![The Myogenic Regulatory Factors 11
D. Axial tissues are required for epaxial somite myogenesis
Cells within the newly formed somite are positionally determined as a result of extrinisic
signals acting during later stages of somite development (Aoyama et al., 1988: Ordaht
et al., 1992). Analysis of the phenotypes of notochord mutant mice and zebrafish suggest
an essential role of the notochord in somite myogenesis (see Figure 3) (for review see
(Borycki et al.. 2000)). These studies along with grafting and explant studies suggest
a combination of signals from the neural tube, notochord and surface ectoderm are
sufficient to induce expression of the MRFs and somitic myogenesis (for review see
IArnold et al.. 1998: Borycki et al.. 2000)). In presomitic mesoderm explants from
mouse 9:5 dpc embryos, axial structures preferentially activate myogenesis through
a Myf5-dependent pathway, whereas axial structure isolated from a 10.5 dpc embryo
preferentially induce MyoD expression independent of Myf5 (Tajbakhsh et al., 1998).
This suggests the temporal separation of Myf5 dependent and independent pathways
is the result of axial tissue maturation (Tajbakhsh et al., 1998). The positive acting
factors include Sonic hedgehog, and Wnt family members (see below, for review see
(Borycki et al.. 2000)).
E. Shh and Wnts
The notochord and floor plate secrete factors including somc hedgehog which
positively regulate myotome formation (for review see (Borycki et al., 2000)).
Sonic hedgehog (Shh) with it two related mammalian proteins Indian hedgehog
and Desert hedgehog are small secreted signaling proteins. It is synthesized in
the notochord, floorplate and zone of polarizing activity in the developing embryo
(Marti et al.. 1995). Shh is required for early activation of Myf5 and MyoD in somite
cells that give rise to the epaxial lineage (see Figure 3) (Borycki et al., 1999). It is
however not essential for the activation of Myf5 and MyoD in the hypaxial dermo-
myotome or the myogenic progenitors that form limb muscle (Borycki et al.. 1999].
The downstream effectors of Shh signaling include Ptc Ithe Shh receptor) and the Gli
transcription factors (for review see (Borycki et al., 200(3)). Although analysis of the
expression patterns of these proteins suggest a possible role in control of myogenesls,
the involvement of molecules downstream of Shh have yet to be determined.
Similarly Wnt-1. Wnt-3 and Wnt-4, which are expressed in dorsal regions of the
neural tu~, can activate myogeneis m vitro (see Figure 3) (Munsterberg et al.. 19957.
Analysis of Wntl +, Wnt3a4- null mice suggests Wnt signals regulate early myogenesis.
in particular early Myf5 expression (Ikeya et al.. 1998). Wnt7a is expressed in
the dorsal ectoderm and preferentially activates MvoD (Tajbakhsh et al.. 1998).
The myogenic promoting activity of Wnt7a is inhibited by BMP4. These results
highlight the role of Shh and Wnt signalling in embryonic myogenesis. In addition°
these results support the hypothesis that the epaxial lineage is Myf5-dependent and
distinct from the MyoD-dependent hypaxial lineage.](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-19-320.jpg)
![16 C. M. Palmer and M. A. Rudnicki
expression of MyoD (Nabeshima et al., 1993). However. analysis of primary MyoD4-
myogenic cells suggest continuous culture of primary myoblasts effects differentiation
potential. The differentiation defect observed in low-passage primary MyoD+ myoblasts
is not observed in later-passage MyoD+ cultures (Sabourin et al., 1999). In fight of
these results, it is important to characterize the differentiation potential of low-passage
primary myogenin + myoblasts.
E. MRF4
The gene encoding mouse MRF4 is linked to the Myf5 gene in a head to tail orientation.
Approximately 8.5 kb of sequence separates the two loci (Miner et at., 1990).
Three laboratories have generated MRF4 mutant mice (Braun et at., 1995; Patapoutian
et aL, 1995; Zhang et al., 1995). Although the identical PGK-neo selection cassettes was
used in the targeting constructs, orientation of the cassette and deleted MRF4 sequence
is unique to each construct [for review see (Olson et at., 1996)]. Analysis of the
role of MRF4 in embryonic myogenesis has been complicated by the close proximity
of the two MRF genes. Targeted disruption of MRF4 affects Myf5 expression via
a cis-acting mechanism tFloss et at., 1996; Yoon et al., 1997). The negative cis
effect of MRF4 mutations on Myf5 transcription was confirmed by the generation
of compound heterozygous MRF4; Myf5 rmce in which the disrupted alleles are on
different chromosomes. These mice have a significant reduction in Myf5 transcript
compared to control animals suggesting the wildtype Myf5 allele is partially inactivated
(Floss et at., 1996; Olson et at., 1996: Yoon et at., 1997). Myf5 enhancer elements
have been identified in a 14 kb region spanning the MRF4/Myf5 locus (Summerbell
et al., 2000). These enhancers regulate Myf5 expression in epaxial muscle precursor
cells, the branchial arches and the central nervous system.
MRF4 null mice generated in Barbara Wold's laboratory die perinatally and show
defects in axial myogenesis and rib structure (Patapoutian et at., 1995). In light of recent
insights into the neo cassette-dependent rib defects observed in some Myf5 mutant
mice, the rib anomalies observed in the MRF4 null mice may be the result of cis effects
on a yet to be identified gene (Kaul et al., 2000). A deficit in myotome development
is observed in the MRF4-/- mice between 9 and 11 dpc. This corresponds to the first
wave of MRF4 expression during development. A significant reduction in Myf5, MyoD
and myogenin expression is observed in mutant mice at day 10 pc. A decrease in
Myf5, MyoD and myogenin levels is also observed in heterozygous mice at this time
(Patapoutian et al., 1995). By 14 pc, myogenesis in these MRF4 mutant animals appears
normal. Minor defects are observed in the intercostal muscle of neonates (Patapoutian
et al., 1995). Although studies using transgenic mice have shown the MRF4 sequences
deleted in the Wold mice are not required for appropriate Myf5 expression, interference
of Myf5 expression by PGK-neo can not be ruled out (Summerbell et al., 2000).
The Olson MRF4 mutant mice are viable however have malformations of the ribs
including bifurcations, fusions and supernumerary processes (Zhang et al., 1995).
Neonatal muscle is grossly normal although the level of embryonicmyosin heavy chain
is slightly decreased. Similar to the Wold MRF4-/-mice, defects in myotome differentia-
tion are observed in 10.5 dpc MRF4-/- embryos (Vivian et at., 2000). Myogenin and](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-24-320.jpg)
![matters occurs in the treasury roll of 1201, where two men in Devon
are set down as owing a fine “because they had been with Count
John”[517]—that is, because they had supported, in his rebellion
against Richard in 1193, the very man for whom, as king, the fine
was now claimed. The Crown had, however, no direct means of
enforcing payment of either fines or taxes, at any rate in the case of
the barons. Its one remedy was to seize the lands or castles of an
obstinate and wilful defaulter; and this remedy was fraught with
danger to the crown itself. Neither law nor custom defined the
circumstances or fixed the limits of time within which a defaulter was
not, and beyond which he was, liable to be treated as obstinate and
wilful; in every case where the king exercised his right of seizure on
this ground, therefore, the defaulter and his friends could always
find a plea for denouncing its exercise as arbitrary and unjust. It
seems probable that at the close of Richard’s reign his ministers may
have thus seized the castles or lands of certain barons in pledge for
the arrears of their dues to the crown, and that this may have been
one of the grievances referred to in the demand of the barons that
Richard’s successor “should restore to each of them his rights.”
John’s demand for the castles of some of the barons in 1201 was in
all likelihood a proceeding of the same kind, based on the same
ground, and, as it seems, equally ineffectual in compelling payment;
all that the king obtained was the surrender not indeed of the
castles, but of some of the barons’ sons as hostages. The deadlock
was probably inevitable; but every year of its continuance
aggravated both the financial difficulties of the government, and the
unfriendliness of the relations between the barons and the king; and
this latter evil was yet further aggravated by the measures which
had necessarily to be taken in order to meet the former one.
Plunged as he was from the very moment of his accession in a costly
struggle with France, John had been forced to lay continually fresh
burdens upon that very class among his subjects who already were,
or considered themselves to be, overburdened by the demands of
his predecessor. The “first scutage of King John” seems to have been
assessed immediately after his coronation; it appears in the Pipe Roll](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-27-320.jpg)
![made up at Michaelmas 1199. In the financial year ending at
Michaelmas 1201, and in every one of the five following years, there
was another new scutage;[518] and these scutages were
independent of the fines paid by the barons who did not accompany
the king on his first return to Normandy in 1199, of the money taken
from the host as a substitute for its service in 1201, of the
equipment and payment of the “decimated” knights in 1205, and the
fines claimed from all the tenants-in-chivalry after the dismissal of
the host in the same year, as well as of the actual services which
many of those who had paid the scutage rendered in the campaigns
of 1202–1204 and 1206.
The other taxes levied during these years were a carucage in
1200[519]and a seventh of moveables in 1204.[520] But all the while
arrears went on accumulating, and year after year a budget had to
be made up by devices of the most miscellaneous character. The
accession of a new king could, of course, easily be made a pretext
for selling confirmations of existing rights and privileges, and John
availed himself of this pretext to the uttermost of his power at the
earliest opportunity—that is, on his visit to England in 1201. During
that time nobody in England seems to have felt secure of anything
that he possessed till he had bought it of the king. Individuals of
various ranks bought the sovereign’s “peace” or his “goodwill”;[521]
the cities of Winchester and Southampton and the county of Hants
each gave him money “that they might be lovingly treated”;[522]
Wiltshire gave him twenty pounds “that it might be well
treated.”[523] The citizens of York offended him by omitting to
welcome him with a procession when he visited their city, and to
provide quarters for his cross-bowmen; he demanded hostages for
their future good behaviour, but afterwards changed his demand to a
fine of a hundred pounds.[524] The sale of offices went on as of old;
[525] while the sale of charters to towns, which under Richard was
already becoming a remarkable item in the royal accounts, was a
transaction of yet greater frequency and importance under his
successor.[526] On the other hand, John’s treasury rolls contain many](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-28-320.jpg)
![1207
notices of persons who owe the king money “which he has lent
them.” These loans from the king to his barons and other subjects
were probably made chiefly in the hope of securing the fidelity of the
borrowers. In one way or another the speculation must have been in
most cases a paying one for John. The privilege of claiming interest
in hard cash for a loan was indeed reserved exclusively for the Jews,
and not shared even by the king; but he could take from his debtors
ample security on their lands or castles, or by means of hostages
who were usually their sons or other young members of their
families, and whom it was of the greater importance for him to hold
in his power as his relations with the barons grew more strained
year by year.
In 1206 the tension had reached such a point
that John did not venture to impose a scutage of
the full amount—two marks on the knight’s fee—which had been
usual since his father’s time, but contented himself with twenty
shillings.[527] In 1207 he evidently dared not attempt to levy any
fresh scutage at all. Nor was a carucage likely to prove either less
unpopular or more productive; for the agricultural interest of the
country was in a state of extreme depression, owing to a long
succession of bad seasons; while the taxation of moveables was an
expedient which seems to have found, as yet, but little favour with
either the people or the government. John now put forth a
suggestion which was, so far as we can see, a novelty in English
finance. He “held a council in London on January 8, and there
requested the bishops and abbots that they would allow parsons and
others holding ecclesiastical benefices to give to the king a fixed sum
from their revenues.”[528] Neither in equity nor in policy was the idea
a bad one. While the military tenants and the socage tenants had
each their own peculiar burden—scutage in the one case, carucage
in the other—the beneficed clergy, as such, had never yet been
subjected to taxation. The king might well argue that it was time for
them to take their turn in making a special contribution to the
financial needs of the State; and the argument was sure to meet
with the approval of the laity. The prelates, however, were unwilling;](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-29-320.jpg)
![and the question was adjourned to another council, in which “an
infinite multitude” of ecclesiastical and temporal magnates came
together at Oxford on February 9.
At this second meeting the bishops of both provinces gave it as
their final answer that “the English Church could by no means
submit to a demand which had never been heard of in all previous
ages.”[529] The only approach to a precedent for it, indeed, had
occurred in 1194, when Archbishop Geoffrey of York, eager to collect
money for Richard’s ransom, had asked the canons of his cathedral
chapter to give for that purpose a fourth part of their revenues for
the year, with the result that they accused him of “wanting to
overthrow the liberties of their church,” and shut its doors in his
face.[530] Between the council in London and that at Oxford,
Geoffrey and John, who had been more or less at variance ever
since the latter’s accession, were formally reconciled;[531] John
therefore probably counted upon Geoffrey’s support of his scheme,
and he may have hoped that the suffragans of Canterbury, having
no metropolitan of their own to lead them, would not venture to
stand out against the northern primate and the king with the barons,
for once, at his back. But what Geoffrey had himself asked of his
own chapter as a special favour to Richard in a wholly exceptional
emergency, he had no mind to give leave for John to claim from all
the beneficed clergy of his province as a matter of right, and under
entirely different circumstances. The king was prudent enough not to
press his demand; but it may be doubted whether the lay barons
agreed with the Waverley annalist in deeming its withdrawal a proof
that he “had taken wiser counsel,” since he substituted for it a
demand for a thirteenth of the moveable goods of every layman
throughout the realm.[532] This they had no excuse for refusing. “All
murmured, but no man dared contradict,”[533] except Geoffrey of
York. He, it seems, claimed exemption for laymen holding lands of
the Church, or at least of his cathedral church. His protest, however,
was disregarded; whereupon he excommunicated all spoilers of the](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-30-320.jpg)
![1208
Church in general, and of the province of York in particular, and then
withdrew over sea,[534] to spend the rest of his life in exile.
Thus for the next eight years the vast diocese of
York was practically without a chief pastor and the
province without a metropolitan, while the temporalities of the see
were in the hand of the king. As for Canterbury, John had answered
the Pope’s request that he would ratify the election of Stephen
Langton by a flat refusal to accept as primate a man of whom he
declared that he “knew nothing, save that he had dwelt much
among his enemies”;[535] and when on June 17 Stephen was
consecrated by Innocent,[536] the king seized the estates of the
Canterbury chapter, drove the monks into exile,[537] and proclaimed
that any one who acknowledged Stephen as archbishop should be
accounted a public enemy.[538] In August Innocent bade the bishops
of London, Ely and Worcester threaten the king, if he continued
obstinate, with an interdict upon his realm, and hinted that this
might be followed by a papal excommunication of John himself.[539]
Negotiations went on throughout the winter, but without result,[540]
and on Passion Sunday, March 23, or Monday, March 24, 1208, the
interdict was proclaimed.[541] It seems that notice of the intended
date of its publication was given about a week before, and that the
king at first answered this notice by ordering all the property of the
clergy, secular or monastic, to be confiscated on Monday, March 24;
but that he immediately afterwards decided to anticipate, instead of
returning, the blow, and caused the confiscation to be begun at
once.[542] For him the opportunity was a golden one. The interdict
enabled him to put the whole body of the clergy in a dilemma from
which there was no escape. They held their property—thus he
evidently argued—on condition of performing certain functions: if
they ceased from those functions, their property was forfeit, just as
that of a layman was forfeit if he withheld the service with which it
was charged. The logical consequence in either case—from John’s
point of view—was confiscation; difficult and dangerous to enforce](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-31-320.jpg)
![1209
on a wide scale against laymen, but easy and safe when the victims
were clergy. The barons made no objection to a proceeding which
would fill the king’s coffers without drawing a single penny from their
own; the chief justiciar himself, Geoffrey Fitz-Peter, earl of Essex,
had no scruple in acting as custos for the Crown of all the Church
property on his own estates, which were scattered through thirty-
one counties, and also of the revenues and goods of the Templars
throughout all England.[543] The spoliation was indeed effected with
a brutal violence which would have been impossible had there been
any strong feeling against it among the influential classes of the
laity,[544] and which so far outran the intentions of the king that on
April 11 he issued a proclamation ordering that any man caught
doing or even speaking evil to a monk or a clerk, “contrary to our
peace,” should be hanged upon the nearest oak.[545] The clergy, like
the Jews, were to be ill-treated by no one save the king himself.
Many of them made a compromise with their spoiler; within a very
few weeks five bishops, three cathedral chapters, the prior of the
Hospitallers, and the heads of fourteen important monasteries,
besides sundry individual priests, undertook to farm their own
benefices and other property for the king.[546] The Cistercians,
asserting that the privileges of their order exempted them from
interdict, ceased from performing the offices of religion for a few
days only, and then resumed them as usual;[547] whereupon their
possessions, which had been seized like those of the other orders,
were restored to them on April 4.[548]
At the same time John despatched an envoy to
Rome proposing terms on which he professed
himself willing to let Stephen take possession of his see; and he
contrived to spin out the negotiations for six months before Innocent
discovered that the terms offered were merely a device for wasting
time, and that the king had never intended to fulfil them.[549] On
January 12, 1209, the Pope informed the bishops of London, Ely and
Worcester that he had written to John a letter of which he sent them
a copy, and bade them excommunicate the king if he did not repent](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-32-320.jpg)
![1208–09
within three months after its receipt.[550] John upon this began a
fresh series of negotiations, which kept the three bishops—who had
apparently gone over sea immediately after publishing the interdict—
flitting to and fro between the continent and England, without any
result, for nine more months. In October they finally withdrew, but
without publishing the excommunication; and by the end of the year
all possibility of its publication in England had vanished, for every
English bishop had fled save two, Peter des Roches, bishop of
Winchester, and John de Grey, bishop of Norwich, both of whom
were creatures of the king; John de Grey, moreover, was now
justiciar in Ireland, and the Poitevin Peter des Roches was thus left
sole representative of the episcopal order in England.[551]
It was John’s hour of triumph, not over the clergy
alone, but over all his subjects and vassals within
the four seas of Britain. The action of the Pope and the inaction of
the barons had opened a way for him to make himself “King of
England” in his own sense of the words. To all outward seeming his
whole time, since his return from the continent, had been devoted to
mere amusement and self-indulgence. He “haunted woods and
streams, and greatly did he delight in the pleasure of them.”[552]
When he was not thus chasing the beasts of the forest, his yet more
relentless pursuit of other prey was making havoc of the domestic
peace, and rousing against him the deadly hatred, of some of the
greatest of his barons.[553] But their hatred was futile; they were
paralyzed partly by their own mutual jealousies, which the king was
continually stirring up,[554] partly by the consequence of their selfish
shortsightedness with regard to his persecution of the clergy. The
interdict had placed one whole estate of the realm at John’s mercy;
and the laity, having failed at the critical moment to make common
cause with their clerical brethren, now found themselves in their turn
without a support against his tyranny. His consciousness of power
broke out in the strangest freaks of wantonness; in causing the
Michaelmas session of the Exchequer to be held at Northampton
instead of London, “out of hatred to the Londoners”;[555] in](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-33-320.jpg)
![forbidding the capture of birds all over England;[556] in ordering that
throughout the Forest districts the hedges should be fired and the
ditches made by the people to protect their fields should be levelled,
“so that, while men starved, the beasts might fatten upon the crops
and fruits.”[557] It showed itself too in acts of graver political
significance. A series of orders to the bailiffs of the coast towns for
the equipment and mustering of their ships and the seizure of
foreign vessels, issued in the spring and summer of 1208, indicates
that John was then either meditating another expedition over sea,
or, more probably, expecting an attack from thence. The muster,
originally fixed for Trinity Sunday, was postponed to S. Matthew’s
day,[558] and the end of the matter was that John, finding he had no
immediate need for the services of the fleet, “took occasion”—no
doubt on pretext of some deficiency in the contingent due from
them—“to oppress the mariners of the Cinque Ports with great and
heavy affliction. Some he hanged; some he killed with the sword;
many were imprisoned and loaded with irons”; the rest fled into
exile, and it was only by giving him fines and hostages that they
appeased his wrath and bought his leave to return to their homes.
[559] The barons were again required to renew their homage; the
demand was made literally at the sword’s point—for John’s lavish
hospitality and largesse[560] filled his court with mercenaries who
were quite ready to enforce his will in such a matter—and they were
compelled either to submit to it, or to give their sons and kinsmen as
hostages for their fidelity.[561] The king seemed indeed, as Matthew
Paris says, to be courting the hatred of every class of his subjects.
[562] But hate him as much as they might, they feared him yet more
than they hated him; and “burdensome” as he was “to both rich and
poor,”[563] when he summoned all the free tenants throughout the
realm, of whatever condition, who were above the age of twelve
years, to swear fealty in person to him and his infant heir in the
autumn of 1209, rich and poor alike durst not do otherwise than
obey him.[564]](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-34-320.jpg)
![1209
Aug. 7
This ceremony took place at Marlborough in
September,[565] just before the final rupture of the
negotiations with Langton and the bishops. A few weeks earlier John
had received the submission of the king of Scots. Twice or thrice in
the last two years a visit of William the Lion to the English court had
been projected.[566] It took place at length in the middle of April
1209 at Bolton, whence John and William proceeded together to
Norham for a conference.[567] The shelter given in Scotland to some
of the bishops and other persons who fled from John’s persecution in
connection with the interdict[568] supplied the English king with a
pretext for demanding, once for all, security for William’s loyalty. He
bade him surrender either three castles on the border or his only son
as a hostage. William refused to do either.[569] John, on returning to
the south, summoned his host, and in July set out to take the three
castles by force. The papal excommunication was hanging over his
head, and its publication was hourly expected; his troops shrank
alike from his leadership and from an encounter with the Scot king,
who was considered “eminent for his piety,” the champion of the
Church and the favourite of Heaven, while they, being under
interdict, were virtually outcasts from the Christian fold. A dexterous
renewal of negotiations with Innocent and Stephen, however, staved
off the excommunication and prevented the threatened desertion of
the English troops;[570] and on August 4 John was at Norham[571] at
the head of a great host ready to do battle with the Scots. On
hearing this, William “greatly feared his attack, knowing him to be
given to every kind of cruelty; so he came to meet him and offered
to treat for peace; but the king of the English flew into a rage and
insulted him bitterly, reproaching him with having received his
(John’s) fugitives and public enemies into his realm, and lent them
countenance and help against him.” At last some “friends of both
realms” arranged terms which pacified John and which William dared
not refuse. He sent his son, not indeed as a
hostage, but to do homage to the English king “for
the aforesaid castles and other lands which he held”;[572] he](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-35-320.jpg)
![1199–1209
undertook to pay John by instalments within the next two years
fifteen thousand marks “to have his goodwill”; he gave hostages for
the fulfilment of this undertaking; and he surrendered his two
daughters to be kept in John’s custody as his wards and married at
his pleasure.[573] According to Gervase of Canterbury, one of these
ladies was to be married to John’s son;[574] one of his many
illegitimate sons must be meant, for though John had now two sons
by his queen, the elder of them was not yet two years old, while the
younger of William’s daughters was thirteen at the least.[575] All that
William obtained in return for these concessions was the freedom of
the port of Berwick, and leave to pull down a castle which the bishop
of Durham had built over against it.[576] Of his claim upon
Cumberland and Westmorland nothing further was ever heard.
Two months later, Wales followed Scotland’s
example. Over Wales, indeed, John’s triumph was
won without the trouble even of a military demonstration on his
part. The anarchy of Wales had been growing worse and worse ever
since the death of Henry II. Its danger for England lay mainly in the
opportunities which it afforded to any of the English barons of the
border who might be treasonably inclined, for making alliances with
one or other of the warring Welsh princes, and thus securing for
themselves a support which might enable them to set at defiance
the authority of the English crown. John himself had held the
position of a border baron for ten years, as earl of Gloucester and
lord of Glamorgan, and had used it for his own private ends as
unscrupulously as any of his neighbours.[577] The familiarity with
Welsh politics which he had thus acquired stood him in good stead
when he became king. At his accession, a struggle which had been
going on for two years between three rival claimants to the
succession in South Wales, Griffith and Maelgwyn, sons of the late
prince Rees ap Griffith, and Gwenwynwyn, son of Owen Cyveiliog,
prince of Powys, had just ended in the triumph of Griffith, who, by
the help of a force supplied to him by the English government,
overcame both his rivals at the close of 1198. On Griffith’s death in](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-36-320.jpg)
![1209–10
1200 Gwenwynwyn for a moment regained the ascendency in South
Wales; but he found a new and formidable rival in the prince of
North Wales, Llywelyn ap Jorwerth, who in a few years succeeded in
reducing most of the South Welsh princes to dependence on himself.
[578] Throughout these years John, amid all his political and military
occupations on the continent, watched every vicissitude of the
struggle in Wales, kept up constant relations with both parties, and
balanced the one against the other[579] with a mingled
unscrupulousness and dexterity for which even the Welshmen were
scarcely a match, and which at last brought them all alike to his feet.
In July 1202 Llywelyn promised to do homage to the English king as
soon as the latter should return from over sea;[580] before October
15, 1204, he was betrothed to John’s illegitimate daughter Joan,[581]
and in 1206 she became his wife.[582] In 1208 his rival
Gwenwynwyn was in an English prison, whence he obtained his
release by doing homage to John at Shrewsbury on October 8.[583]
Llywelyn’s promised visit to the English court seems to have not yet
taken place; but a year later, on the king’s return from the north,
there befell, say the chroniclers, “what had never been heard of in
times past: all the Welsh nobles”—that is, evidently, the princes of
both North and South Wales—“came to him and did him homage,”
not on the border, but in the heart of his own realm, at Woodstock,
[584] on October 18 or 19, 1209.[585]
The king’s triumph was complete. The last date
which had been fixed for the publication of the
papal sentence was October 6;[586] the sentence was still
unpublished, and the bishops who should have published it had fled.
They proclaimed it indeed in France in November;[587] but John took
care that no official notification of the fact should reach England,
and the sentence remained a dead letter. Its existence was known
and talked of all over the country, but it was talked of with bated
breath. The excommunicate king held his Christmas feast at Windsor
surrounded by “all the great men of England,” who sat at his table](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-37-320.jpg)
![and held intercourse with him as usual, simply because they dared
not do otherwise.[588] Of the fate in store for those who stood aloof,
one terrible example sufficed. The archdeacon of Norwich quitted his
place at the Exchequer table at Westminster, after warning his
fellow-officers that they were perilling their souls by serving an
excommunicate king. He was seized by a band of soldiers, loaded
with chains, flung into prison, and there crushed to death beneath a
cope of lead.[589] The whole body of the clergy, already stripped of
their possessions, were now in peril of their lives. As the king was
passing through one of the border counties he met some of the
sheriff’s officers in charge of a prisoner with his hands tied behind
him. They said the man was a robber, and had robbed and slain a
priest on the highway: what, they asked, should be done with him?
“Loose him and let him go” answered John, “he has slain one of my
enemies!” Nor was his persecution limited to the clergy; the lay
relatives and friends of Langton and of the other exiled bishops were
hunted down and flung into prison, and their property seized for the
king.[590] When he could plunder his Christian subjects no more, he
turned upon the Jews. At the opening of 1210 all the Jews in
England, of both sexes, were by his order arrested, imprisoned, and
tortured to make them give up their wealth. It was said that the king
wrung ten thousand marks from one Jew at Bristol by causing seven
of his teeth to be torn out, one every day for a week,[591] and that
the total sum transferred from the coffers of the Jews to the royal
treasury amounted to sixty-six thousand marks.[592] Never before—
not even in the worst days of William the Red—had England fallen so
low as she now lay at the feet of John. “It was as if he alone were
mighty upon earth, and he neither feared God nor regarded
man.”[593] John seems in fact to have been one of the very few men
of whom this latter assertion can be made with literal truth; and in
this utter recklessness and ruthlessness lay the secret of his terrible
strength. “There was not a man in the land who could resist his will
in anything.”[594] The very few barons who had dared openly to](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-38-320.jpg)
![1191–1200
resist it since his return from Poitou in 1206 were now all in Ireland;
and it was Ireland that he set himself to subdue in 1210.
John de Courcy had apparently ceased to be
governor of the Irish March in 1191. The succession
of governors there during the next few years is obscure; but we
know that, as John’s chief ministers, they bore the same title which
was borne by the chief minister of the king in England, that of
justiciar.[595] Owing to the paucity and obscurity of the records it is
difficult to gain any real understanding of the vicissitudes of the
English dominion in Ireland during the twenty-five years which
elapsed between John’s two visits to that country, and especially
during the fourteen years between his first visit there and his
accession to the English crown. He granted a new and important
charter to the city of Dublin in 1192.[596] In 1195 the intruders—
neither for the first nor for the last time—fell out among themselves:
“John de Courcy and the son of Hugh de Lacy marched with an army
to conquer the English of Leinster and Munster.”[597] They certainly
did not succeed in wresting Leinster from William the Marshal. As for
Munster, Richard de Cogan was apparently still holding his ground in
Desmond; Raymond the Fat probably died in 1184 or 1185,[598] and
as he had no direct heirs,[599] the share of that kingdom which had
been originally allotted to Fitz-Stephen lapsed to John as overlord.
[600] From the city of Cork the “English” are said to have been driven
out in 1196;[601] but their expulsion was only momentary.
Meanwhile they had at last begun to gain a footing in Thomond. By
1196 they had got possession of the city of Limerick; in that year or
the next they lost it, but it was speedily recovered by Meiler Fitz-
Henry,[602] who in 1199 or early in 1200 became chief justiciar in
Ireland.[603] Limerick was put under the charge of William de Burgh,
who apparently had won for himself some lands within the kingdom
of Thomond, among them Ardpatrick, of which he received a grant
from John in September 1199.[604]](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-39-320.jpg)
![1198–1202
1179–1201
The last Irish Ard-Righ, Roderic O’Conor, died in
1198;[605] he had been dethroned sixteen years
before, but his death was the signal for renewed strife between his
sons for the possession of his kingdom of Connaught. The foreign
settlers in Ireland took sides for their own interest in the struggle
between the native princes; John de Courcy and the “English of
Ulidia,” with the De Lacys of Meath and their followers, supported
Cathal Crovderg O’Conor, while his rival, Cathal Carrach, was helped
by “William Burke, with the English of Limerick.” For a moment
Cathal Carrach’s party was victorious; but next year (1200) he was
attacked by “Meiler and the English of Leinster,” while De Burgh
changed sides and joined Cathal Crovderg. In 1201 or 1202 the
united forces of Cathal Crovderg and De Burgh won a battle in which
Cathal Carrach was slain. Cathal Crovderg being thus master of
Connaught, De Burgh at once began to plot against his life; but the
men of Connaught slaughtered the followers of the double-dyed
traitor, and he himself escaped as best he could back to Limerick.
[606]
The “honour of Limerick”—exclusive of the city
and the Ostmen’s cantred, which the king retained
in his own hands, and the service due from the lands held within
that honour by William de Burgh, which was also reserved to the
Crown—had meanwhile been granted by John, on January 12, 1201,
to William de Braose, “as King Henry gave it to his uncle, Philip de
Braose.”[607] These last words define the extent of the “honour,” as
corresponding (with the exceptions specified) to the “kingdom of
Limerick” (Thomond) named in Henry’s grant of 1177. Philip de
Braose was probably now dead. William was the son of Philip’s elder
brother, another William who to the family estates of Bramber in
Sussex and Barnstaple and Totnes in Devon had added, by his
marriage with an heiress, the lordships of Radnor, Brecon, and
Abergavenny in Wales.[608] The younger William probably succeeded
to all these possessions soon after 1179.[609] Before 1189 his sister
Maud was married to Griffith Ap Rees, who from 1198 to 1201 was](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-40-320.jpg)
![1201–1204
Prince of South Wales; and throughout the last ten years of the
twelfth century William was constantly concerned in the quarrels of
the South Welsh princes and people.[610] His daughter Margaret had
before November 19, 1200 become the wife of Walter de Lacy,[611]
the lord of Meath, who was already her father’s neighbour on the
Welsh border, where Ludlow formed part of the Lacy heritage; a
younger daughter was married before 1210 to a son of another
baron of the Welsh March, Roger Mortimer.[612] Count John of
Mortain, as earl of Gloucester and lord of Glamorgan, was also for
ten years a neighbour of William de Braose, and evidently made a
friend of him, for in 1199 William was at the head of the party which
most vigorously urged John’s claim to the crown.[613] In June 1200
he received a royal grant of “all the lands which he had acquired or
might at any future time acquire from our Welsh enemies, to the
increase of his barony of Radnor.”[614] As the king was at the same
time in diplomatic relations with several of the “enemies” whom
William was thus authorized to despoil, this grant was of doubtful
value. The same may be said of the grant of Thomond; this,
however, was a speculation on both sides; William covenanted to
pay the king five thousand marks for it at the rate of five hundred
marks a year.[615]
De Braose immediately went to Ireland;[616] and
in process of time he succeeded in obtaining
possession of the greater part of his new fief, though the difficulties
with which he had to contend were many and great. The other
persons who had previously received from John grants of land in
Thomond[617] no doubt resented and resisted the change in their
position from tenants-in-chief of the king to under-tenants of William
de Braose. It seems that they were upheld in their resistance by the
justiciar, Meiler Fitz-Henry, and that John in consequence summoned
Meiler to his court, suspended him from his office, and put it into
commission in December 1201. In August 1202 John issued further
orders for enforcing the claims of De Braose in Thomond; in
September he forgave him all the debts which he owed to King](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-41-320.jpg)
![1204–1206
Henry and King Richard; in October he granted the entire custody of
the lands and castles of Glamorgan, Gwenllwg and Gower to
“William de Braose, whose service we greatly approve.”[618] In the
winter William was with the king in Normandy, and had the custody
of the captive Arthur. This he resigned, seemingly at the end of the
year,[619] and in January 1203 he was in charge of some matters
connected with the fleet.[620]
Meanwhile the governor of Limerick city, William
de Burgh, had escaped from the vengeance of the
Irish allies whom he had betrayed, only to fall under that of the
English justiciar whom he had set at defiance. Meiler Fitz-Henry had
been restored to his post; in 1203 he and Walter de Lacy joined with
the Irish of Connaught in expelling De Burgh from Limerick,[621] and
on July 8 William de Braose was appointed by the king to succeed
De Burgh as constable of the city.[622] Meiler and De Burgh had
already appealed against each other to the king;[623] in March 1204
a commission was appointed to hear their reciprocal complaints;[624]
in September all De Burgh’s Irish estates except those in Connaught
were restored to him on his promise of “standing to right in the
King’s Court of Ireland.”[625] There is no record of the trial, which
may have been prevented by his death, for at the end of the year or
in 1205 he died;[626] and on April 3, 1206 the justiciar was ordered
to take all his Munster estates into the king’s hand.[627]
The reservation of De Burgh’s Connaught lands in 1204 may have
been made in consequence of some negotiations which were at that
moment going on between Meiler, as John’s representative, and the
King of Connaught, Cathal Crovderg. Cathal, it seems, offered to
cede two-thirds of Connaught to John, on condition that the
remaining third should be secured to himself and his heirs for a
yearly payment of one hundred marks. John was willing to accept
this offer, but he insisted that the portion of land to be ceded to him
should be chosen by Meiler, and bade Meiler take care that it was
“the best part, and that which contained the best towns, ports, and](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-42-320.jpg)
![1199–1205
sites for castles.”[628] Possibly this claim of John’s to choose the land
for himself was refused by Cathal; the negotiations certainly came to
nothing, for in December 1206 Cathal made another proposition. He
would hold one-third of Connaught of King John for a hundred
marks a year; out of the other two-thirds he would cede to John two
cantreds, and for the remainder he would pay him a tribute of three
hundred marks. John authorized Meiler to accept these terms, if he
could get no better.[629] Whether the agreement was ever actually
made, there is nothing to show; it was not likely to have any
practical result. The invaders had evidently already gained some
slight and precarious footing in eastern Connaught; but they had too
much to do within their own March—as the dominions of the English
crown in Ireland were called in those days[630]—to make any real
progress westward for some years to come.
The turbulence and lawlessness which prevailed
in the Irish March reflected that of the Welsh March
whence most of its original settlers had come. William de Braose and
William de Burgh were far from being the only barons at feud with
Meiler Fitz-Henry, either simply as a fellow-baron, or in his official
capacity of representative of the king. In September 1199 John de
Courcy and Walter de Lacy are mentioned in a royal writ as having
acted together “for the destruction of our realm of Ireland.”[631] The
reference probably is to their joint attack upon Leinster in 1195,
which had been followed by the forfeiture of Lacy’s English and
Welsh lands; these, however, he had regained in 1198.[632] In 1203,
as has been seen, he helped Meiler to expel William de Burgh from
Limerick; and in February 1204 he was appointed one of four
commissioners to assist Meiler in dealing with escheats.[633] His
former ally, John de Courcy, had a safe-conduct to and from the
king’s court in July 1202;[634] but he evidently did not come to terms
with the king; and next year the Lacys turned against him; Hugh de
Lacy, Walter’s younger brother, defeated him in a battle near Down
and drove him out of Ulidia.[635] In September he had another safe-
conduct to go to the king and return “if he does not make peace](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-43-320.jpg)
![1204
with us.”[636] This time it seems that he did “make peace,” but failed
to fulfil its conditions. On August 31, 1204, he was summoned, on
pain of forfeiture, to come to the king’s service “as he swore to
come”; and Meiler was instructed, if the forfeiture should take place,
to give to the two De Lacys the eight cantreds of De Courcy’s land
which lay nearest to Meath.[637] De Courcy incurred the forfeiture;
Meiler seemingly committed its execution to the De Lacys; they
again attacked De Courcy, and drove him to take refuge in Tyrone;
[638] and on May 2, 1205, King John granted Ulster to Hugh de Lacy,
to hold “as John de Courcy held it on the day when Hugh defeated
him.”[639] A few weeks later Hugh was belted earl of Ulster;[640] and
at the end of June the triumph of the Lacys was completed by a
royal order forbidding the chief justiciar to “move war against any
man of the March” without the consent of Earl Hugh and his brother
Walter.[641]
With the colleagues thus forced upon him Meiler
was soon at strife. His strife with Walter de Lacy,
indeed, had recommenced already. Walter’s appointment as a
commissioner of escheats in 1204 had been made in connexion with
a demand which John—anxious to prepare for an attack upon
France, as well as to guard against an expected French invasion of
England, and scarcely daring to ask his English subjects for more
money—addressed to all his vassals in Ireland, that they would
furnish him with an aid.[642] They undertook to do so; on September
1 the king thanked them for their services and their promises, and
desired that the latter might be fulfilled.[643] At the same time he
was taking measures for the security of the March and of his own
authority there; on August 31 he had ordered Meiler to build a castle
at Dublin,[644] and in September he bade the citizens do every man
his part in helping to fortify the city.[645] In November he decided
upon taking back into his own hands the city of Limerick and its
cantred, being, as he said, advised by his barons of England that this
step was necessary for the security of his domains in Connaught and](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-44-320.jpg)
![1207 March
1207 April
1207
Cork. It appears that William de Braose had called in the help of his
son-in-law, the lord of Meath, for the keeping of this important
border-post; the king’s orders for its surrender to the justiciar were
addressed to Walter de Lacy and the bailiffs of William de Braose.
[646] Walter seemingly refused to obey the order; Meiler, however,
succeeded in taking possession of the city, “on account of which
there arose a great war” between him and De Lacy,[647] with the
result that John, to end their strife, took away the custody of
Limerick from both of them, and restored it in August 1205 to
William de Braose.[648] Nineteen months later Walter de Lacy’s
castle of Ludlow was seized for the Crown, and
Walter was bidden to come and “stand to right” in
the English court.[649]
By that time Meiler was at strife with William de
Braose again, and also with another Marcher lord of
very different character from any of those with whom he had as yet
had to deal. Meiler Fitz-Henry, though loyal to the king, was
evidently not quite the man for the post of chief justiciar in Ireland.
He was one of the few survivors of the first band of Norman-Welsh
adventurers who had taken part in the invasion under Robert Fitz-
Stephen. The royal blood of England and of Wales was mingled in
his veins; he was in fact, though not in law, first cousin to Henry II.
[650] The two young Lacys, now so often opposed to him, were
cousins of his wife, a niece of the elder Hugh de Lacy.[651] He was,
however, not one of the great barons of the March; he seems to
have held in chief of the Crown nothing except three cantreds in
Desmond granted to him by John in October 1200;[652] his principal
possession was the barony of Leix in Ossory,[653] for which he owed
homage to William the Marshal as lord of Leinster. In the spring of
1207 William the Marshal asked leave of John to visit his Irish lands,
which he had never yet seen. The leave was given, though
unwillingly; but as William was on the point of setting out from
Striguil, he was overtaken by a message from the king, bidding him
either remain in England, or give his second son as a hostage.](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-45-320.jpg)
![1207 Feb. 12
William sent the boy back with the messenger, saying that the king
might have all his children as hostages if he pleased,[654] but as for
himself, he was determined to go to Ireland; and next day he sailed.
His coming was far from welcome to the justiciar, who till then had
been without a superior in the country, and who resented alike the
necessity of doing homage to the Marshal for the land which he held
under him, and the probability of his own importance being
overshadowed by the presence of a man whose territorial and
personal weight was so much greater than his own. Meiler therefore
wrote to the king urging him to recall the Marshal. John did so, but
bade Meiler himself come over at the same time. The Marshal,
though feeling that mischief was in prospect, obeyed the king’s
summons with his usual readiness, and returned to England at
Michaelmas, leaving his wife with a band of trusty followers to
defend Leinster in his stead. Meiler also came, after secretly bidding
his kinsmen and friends attack the Marshal’s lands as soon as he was
gone, which they did the very next week. The king gave Meiler a
warm welcome, but treated the Marshal with coldness and
displeasure,[655] which Meiler soon found a way to increase.
At the beginning of the year the justiciar had seized for the Crown
some of the lands, men and goods of William de Braose.[656] His
excuse for this proceeding was probably the fact that De Braose was
in debt to the Crown for the ferm of the city of Limerick, and also for
no less than four thousand two hundred and ninety-eight marks of
the five thousand which he had in January 1201 covenanted to pay,
by instalments of five hundred every year, for the grant of the
honour of Limerick.[657] Meiler, however, had acted without
instructions from the king; and when De Braose complained of the
treatment which he had received, John declared
that he “found no fault in him,” and bade Meiler
restore everything that had been taken from him, unless indeed the
city of Limerick was included; if that had been seized for the Crown,
Meiler was to retain it till further orders.[658] The mingled feelings of
the king are reflected in his letter. John had found in William de](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-46-320.jpg)
![1207–08
Braose a useful servant and friend; he knew that he might find in
him a dangerous enemy; he was therefore reluctant to take any
measures which might drive William into opposition. On the other
hand, William’s neglect of his pecuniary obligations to the Crown had
reached such a pass that it could hardly be ignored much longer;
and William was further suspected of being in secret alliance against
the king, both with the Welsh and with the De Lacys.[659] Of this
suspicion the king seems to have known nothing till after the middle
of July, when he reappointed “our beloved and faithful William de
Braose” custodian of Ludlow Castle.[660] It had, however, reached
his ears by the time of Meiler’s coming to England, and Meiler turned
it to account for a double purpose of his own. One day, as the king
and his chief counsellors sat talking together after dinner, something
was said about William the Marshal and his friendly relations with
William de Braose. Meiler wrought upon the king’s jealousy of the
one and his suspicions of the other, till he persuaded him to join in a
plot for bringing them both to ruin.
At the justiciar’s instigation John secretly
despatched letters to all those of the Marshal’s
followers in Ireland who held lands in England, bidding them, on
pain of forfeiting these, to be at his court within a fortnight. At the
same time Meiler, with the king’s licence, returned to Ireland. The
Marshal asked permission to do the same; but this was refused.
Meiler on his arrival found that hitherto his men had, on the whole,
been worsted in their strife with those of Leinster. He now
summoned the Marshal’s men to a “parliament,” at which the king’s
messenger read out the secret letters. The men to whom these
letters were addressed saw but too plainly what would be the result
of their obedience: the Marshal’s lands would be left without defence
against Meiler. They unanimously resolved to sacrifice their own
English estates, disobey the king for their lord’s sake, and resist
Meiler to the uttermost; and with the help of two powerful
neighbours whom they called to their aid, Ralph Fitz-Payne and
Hugh de Lacy, they succeeded, as one of them says, in doing to](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-47-320.jpg)
![1208–09
Meiler as much mischief as he had thought to do to their lord.[661]
The Marshal, meanwhile, was compelled to remain at court, but so
discountenanced by the king that hardly any one dared to speak to
him. At last, one winter day, as they rode out from Guildford,[662]
John called to him: “Marshal, have you had any news from Ireland
that pleases you?” “No, sire.” “I can tell you some news,” said the
king, laughing; and he told him that his wife, the Countess Isabel,
had been besieged in Kilkenny by Meiler, who had indeed been at
length worsted and even captured by her people, but with very
heavy losses on her side, three of the Marshal’s chief friends being
among the slain. The story was a sheer invention of John’s; in reality
he had received no news from Ireland at all. The Marshal, though
perplexed and troubled, retained his outward composure; and early
in the spring he himself received from Ireland a very different
account of what had happened there. The justiciar had not only
been captured, but had made submission to the countess and given
his son as a hostage till he himself should stand to right in her
husband’s court for the wrong which he had done to him as his lord.
These tidings were sent at the same time to the
king, who was by no means pleased with them, but
characteristically changed his policy at once to meet the turn of the
tide. He called the Marshal to his presence, greeted him with
unusual courtesy, and asked him if he had heard anything from
Ireland. “No, sire; I have no news from thence.” “Then I will tell you
some good news, of which I wish you joy”—and thereupon John
related the truth, which William knew already, though he had not
chosen to say so. From that time forth “the king made him as good
cheer as he had made him evil cheer before”; and when the Marshal
soon afterwards again asked leave to go to Ireland, it was granted at
once.[663] On March 7 Meiler was ordered to refrain from interfering
with the lands of the Marshal, who had instructed his men to keep
the peace towards Meiler in return;[664] on March 20 John informed
the justiciar that “the Marshal has done our will,” and despatched to
Ireland four commissioners by whose instructions Meiler was to act,](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-48-320.jpg)
![1208
and who, if he failed to do so, were empowered to act in his stead.
[665] On the 28th, a new grant of Leinster, on the terms of the
original grant to Richard de Clare, was made by the king to the
Marshal.[666] A month later Meath was in like manner granted afresh
to Walter de Lacy;[667] and at the end of the next year, 1209, Meiler
was removed from his office of justiciar, and replaced by the bishop
of Norwich, John de Grey.[668]
On one point, however, Meiler was justified by
the king. In the spring of 1208 John made up his
mind to bear with William de Braose no longer, and ordered a
distraint upon his Welsh lands. William’s wife, Maud of Saint-Valery,
[669] his nephew, Earl William of Ferrars, and his sister’s husband,
Adam de Port, met the king at Gloucester and persuaded him to
grant an interview to William himself at Hereford. William promised
to pay his debts to the treasury within a certain time, pledged some
of his castles for the payment, and gave three of his grandsons and
four other persons as hostages.[670] Roger of Wendover relates that
when the king’s officers went to fetch the hostages, Maud refused to
deliver up her grandchildren to the king, “because,” said she, “he
has murdered his captive nephew”; that her husband reproved her,
and declared himself willing to answer according to law for anything
in which he had offended the king; and that John, on hearing what
Maud had said, was “greatly perturbed,” and ordered the whole
family of De Braose to be arrested.[671] John himself, in a public
statement attested by the chief justiciar of England and twelve other
men of high position, among whom were De Braose’s own nephew
and brother-in-law, asserted that shortly after the meeting at
Hereford De Braose and his sons attempted to regain the pledged
castles by force, and when they had failed in this attempt, attacked
and burned Leominster.[672] Thereupon it seems that William was
proclaimed a traitor; on September 21 John empowered Gerald of
Athies to make an agreement with all who were or had been](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-49-320.jpg)
![1209–10
homagers of William de Braose, so that they should “come to the
king’s service and not return to the service of William.”[673]
V.
Stanford’s Geogˡ. Estabᵗ. London.
London: Macmillan & Co., Ltd.
De Braose was chased by the king’s officers,[674]
till in the following year, 1209, he escaped, with his
wife and two of their sons, from some Welsh seaport, intending to](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-50-320.jpg)
![go to Ireland. A violent storm kept them tossing on the sea for three
days and three nights; at last they landed at Wicklow. William the
Marshal chanced to be there; he received them kindly and sheltered
them for three weeks. Then their presence was discovered by the
new justiciar, Bishop John de Grey, who at once taxed the Marshal
with harbouring “the king’s traitors,” and bade him give them up to
justice. The Marshal refused, saying he had only received “his
lord,”[675] as he was bound to do, and without knowing that De
Braose had incurred the king’s displeasure; and he added that he
himself would not act like a traitor towards De Braose at the
justiciar’s bidding. Thereupon he sent the refugees safely on to their
destination, the home of De Braose’s son-in-law, Walter de Lacy. The
justiciar complained to the king, who summoned his host for an
expedition to Ireland;[676] both the Marshal and the Lacys having
positively refused to give up De Braose, though they offered to be
answerable for his going to England to satisfy the king within a fixed
time, and promised that, if he failed to do so, they would then
harbour him no more. At last—seemingly in the spring of 1210—De
Braose was allowed to go on these conditions back to Wales. John
had apparently consented to meet him at Hereford; but when De
Braose reached Hereford, “he,” says the king, “regarded us not,” but
began to collect all the forces he could muster against the Crown.
His nephew, the earl of Ferrars, however, managed to bring him to a
meeting with the king at Pembroke. He offered a fine of forty
thousand marks. “We,” says John, “told him we knew well that he
was not in his own power at all, but in that of his wife, who was in
Ireland; and we proposed that he should go to Ireland with us, and
the matter should be settled there; but he chose rather to remain in
Wales,”[677] and was suffered to do so—John being determined now
to settle matters not only with Maud de Braose, but with all the
barons of the Irish March, according to his own will and pleasure.
At some date between June 16 and 20 John crossed from
Pembroke to Crook, near Waterford. Thence he proceeded by way of
Newbridge and Thomastown to Kilkenny, where he and all his host](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-51-320.jpg)
![1210
were received and entertained for two days (June 23 and 24) by
William the Marshal.[678] On June 28 the king reached Dublin;
thence he led his host into Meath.[679] Walter de Lacy and the De
Braoses fled, evidently into Ulster; thither John marched in pursuit of
them, but before he could overtake them they had escaped over sea
into Galloway.[680] Hugh de Lacy had retired into the stronghold of
Carrickfergus; at the king’s approach, however, he, too, slipped away
in a little boat to Scotland.[681] Carrickfergus was provisioned for a
siege, but its garrison was soon frightened into surrender.[682] While
John was at Carrickfergus, his “friend and cousin,” Duncan of
Carrick, sent him word that he had captured Maud de Braose, one of
her daughters, her eldest son, his wife and their two children; her
younger son, Reginald, had escaped, and so had the Lacys. The king
despatched John de Courcy (whom he had taken back into favour,
and brought with him to Ireland, as likely to be a willing and useful
helper against the De Lacys) to fetch the captives from Galloway.
When they were brought before him, Maud offered the surrender of
all her husband’s lands and a fine of forty thousand marks, which
John accepted; but three days later she repudiated her agreement.
[683] Taking his prisoners with him, the king turned southward again,
and soon completed the subjugation of the Lacys’ territories. Most of
the lesser barons fled before him as their lords had done, “fearing to
fall into his hands.”[684] A week’s stay in Dublin (August 18 to 24)
brought his expedition to a close.[685]
It was probably during this second stay of John’s
at Dublin that, as Roger of Wendover says, “there
came to him there more than twenty kinglets[686] of that country,
who all, terrified with a very great fear, did him homage and fealty;
yet a few kinglets neglected to come, who scorned to do so, because
they dwelt in impregnable places. Also he caused to be set up there
English laws and customs, establishing sheriffs and other officers
who should judge the people of that realm according to English
laws.”[687] This latter statement of Roger’s may have given rise to](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-52-320.jpg)
![the later belief that it was John who organized the administration of
the March in Ireland after the English model, by dividing the whole
of the conquered territory into counties, each under its own sheriff.
[688] It appears, however, that there were sheriffs in Ireland in the
days of Henry II.[689] The earliest known mention of a sheriff’s
district there occurs in 1205, when we hear of the “county of
Waterford.”[690] Ten years later the same county is mentioned again,
and also that of Cork;[691] and before the end of the century ten
counties, at least, were recognized by the English government in
Ireland.[692] The names of the earliest Irish counties thus known to
us and the circumstances of John’s visit to Ireland in 1210 may
suggest a clue to the rise and growth of the shire-system in that
country. The district which forms the present county of Waterford
had never been enfeoffed either by Henry II. or by John, but
remained directly in the hands of the supreme ruler of the March. Of
the present county Cork, the eastern half, at least, escheated
together with the rest of Raymond FitzGerald’s share of the
“kingdom of Cork” on his death about 1185. No notice of a new
enfeoffment of any of the lands which had been his occurs till 1208,
and then they were not granted as a whole; so far as we know, only
a portion of them was enfeoffed, and that portion was distributed
among several feoffees.[693] It seems probable that the system of
county administration may have been first established in Ireland in
those districts which were under the direct rule of the English Crown
(or, to speak more exactly, of the “English,” or Angevin, “Lord of
Ireland”), and of which the continuous extent was too great for
them to be left, like the single cantreds attached to the other
seaport towns, under the control of a mere military governor or
constable, and that it was only by degrees introduced into the great
fiefs. If this were so, the events of 1210 would furnish an excellent
opportunity for its extension. Of the four great fiefs which, together
with the royal domains and the lately redistributed honour of Cork,
made up the “English” March in Ireland, Leinster was, when John
sailed from Dublin for England at the end of August,[694] practically](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-53-320.jpg)
![the only one left. Meath, Ulster, and Limerick were all forfeit to the
Crown; and the Crown kept the greater part of them for many years
after. Meath was not restored to Walter de Lacy till 1215;[695]
Walter’s brother, the earl of Ulster, did not return from exile till after
John’s death;[696] and the honour of Limerick was never again
bestowed as a whole upon a single grantee. Under these
circumstances a system of administrative division into counties
placed under sheriffs appointed by the king, or by the justiciar in his
name, might be established without difficulty in territories where its
introduction in earlier years, if ever attempted, would probably have
been rendered ineffectual by the power of the great barons. The one
great baron who in the autumn of 1210 still held his ground in the
March—Earl William the Marshal, the lord of Leinster—had no
hesitation in withstanding the king to his face in the cause of honour
and justice; but he was not a man to throw obstacles in the way of
the royal authority when it was exercised within the sphere of its
rights and in the interest of public order.
On the king’s return to Dublin William the Marshal came to the
court. John at once accused him of having “harboured a traitor” in
the person of William de Braose. The Marshal answered the king as
he had answered the justiciar, and added that if any other man
dared to utter such a charge against him, he was ready to disprove
it there and then. As usual, no one would take up his challenge;
nevertheless, John again required hostages and pledges for the
Marshal’s fidelity, and again they were given at once.[697]
Meanwhile, the sheriff of Hereford sent word that William de Braose
was stirring up trouble in Wales, and urged that he should be
outlawed; but the king ordered that the matter should await his own
return to England. When he was about to sail, Maud de Braose
offered to fine with him for forty thousand marks, and ten thousand
in addition, as amends for having withdrawn from her former
agreement. John accepted these terms; the fine was signed and
sealed, and it was agreed that Maud, and also, it seems, the other
members of her family who had been captured with her, should](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-54-320.jpg)
![remain in custody till it was paid. John carried his prisoners back
with him to England, put Maud in prison at Bristol, and at her
request gave an audience to her husband, who ratified the fine
which she had made, but fled secretly just before the day fixed for
paying the first instalment. The king asked Maud what she now
proposed to do, and she answered plainly that she had no intention,
and no means, of paying. Then it was ordered that “the judgement
of our realm should be carried out against William,” and he was
outlawed.[698] Thus far the king tells his own story, and there is no
reason to doubt its truth. What he does not tell is the end of the
story. He sent Maud and her son to a dungeon at Windsor, and there
starved them to death.[699]
FOOTNOTES: [Skip footnotes]
[502] July 15–20, 1205, Itin. a. 7.
[503] Gerv. Cant. vol. ii. p. 98. Cf. Rot. Pat. vol. i. p. 60, 60 b.
[504] Cf. Innoc. III. Epp. l. viii. No. 161, and Gerv. Cant. l.c.
[505] R. Wendover, vol. iii. p. 183. Cf. Gerv. Cant. vol. ii. p. 99.
[506] Innoc. III. Epp. l. viii. No. 161.
[507] R. Wendover, vol. iii. pp. 184, 185.
[508] Rot. Pat. vol. i. p. 56 b.
[509] R. Wendover, vol. iii. p. 185; Rot. Pat. l.c.
[510] Innoc. III. Epp. l. ix. Nos. 34, 35, 36.
[511] Rot. Pat. vol. i. pp. 65 b, 67.
[512] Ib. p. 64.
[513] M. Paris, Hist. Angl. vol. ii. p. 111; Chron. Maj. vol. ii. p. 514. Cf.
W. Coventry, vol. ii. pp. 197, 198.](https://image.slidesharecdn.com/3781-250605204938-7379c840/85/Stem-Cells-and-Cell-Signalling-in-Skeletal-Myogenesis-1st-Edition-D-A-Sassoon-55-320.jpg)
Stem Cells and Cell Signalling in Skeletal Myogenesis 1st Edition D.A. Sassoon
- 1. Stem Cells and Cell Signalling in Skeletal Myogenesis 1st Edition D.A. Sassoon download pdf https://ebookultra.com/download/stem-cells-and-cell-signalling-in- skeletal-myogenesis-1st-edition-d-a-sassoon/ Visit ebookultra.com today to download the complete set of ebook or textbook!
- 2. We have selected some products that you may be interested in Click the link to download now or visit ebookultra.com for more options!. Stem Cells in Ophthalmology 1st Edition Daniel H. Scorsetti https://ebookultra.com/download/stem-cells-in-ophthalmology-1st- edition-daniel-h-scorsetti/ Stem Cells and Regenerative Medicine Philippe Taupin https://ebookultra.com/download/stem-cells-and-regenerative-medicine- philippe-taupin/ Embryonic Stem Cells 1st Edition Irina Klimanskaya https://ebookultra.com/download/embryonic-stem-cells-1st-edition- irina-klimanskaya/ Stem Cells in Regenerative Medicine 1st Edition Alexander Meissner Phd https://ebookultra.com/download/stem-cells-in-regenerative- medicine-1st-edition-alexander-meissner-phd/
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- 5. Stem Cells and Cell Signalling in Skeletal Myogenesis 1st Edition D.A. Sassoon Digital Instant Download Author(s): D.A. Sassoon ISBN(s): 9780444506634, 0444506632 Edition: 1 File Details: PDF, 21.88 MB Year: 2002 Language: english
- 7. GENERAL OVERVIEW OF THIS VOLUME Skeletal muscle development is perhaps one of the best understood processes at the molecular, cellular and organismal level due in large part to the fact that primary myogenic cells Imyoblasts) will grow and subsequently differentiate into myotubes in culture. This experimental advantage which was pioneered over 30 years ago, coupled with intense efforts to elucidate the causes of multiple muscle myopathies has lead to the identification of growth factors, transcription factors and cell-cell interactions that control this process. With the advent of reverse mouse genetics, many of the observations gained through the study of myogenic cells in vitro have been directly tested in vivo. What has emerged is a complex but cohesive story of how myogenic cells are initially specified in the vertebrate embryo and how muscle fibers ultimately achieve their respective identities (i.e. fast versus slow) to perform their function. While this collection of chapters is focused on these developments, it should be noted that the general paradigms that have emerged have been useful in the study of other tissue types. Skeletal muscle, like nerve cells, undergoes a permanent exit from the cell cycle upon differentiation, and it is generally thought that myofibers are long lived cells. One of the early questions in the field which came from an interest in stimulating regeneration of diseased muscle was where are the myogenic stem cells in skeletal muscle tissue? The identification of the satellite cell appeared to resolve this issue. These cells which lie under the basal lamina (cell matrix) of a myotube with highly condensed nuclei reflecting a quiescent state were observed m enter the cell cycle and reform myotubes. While these cells were identified about 30 years ago. it has been only in the last 5 years that the molecular processes underlying their regulation have become identified. Work by Rudnicki and colleagues has shown that MyoD is critical for satellite cell function and functions downstream of a patterning gene called pax7. One obstacle faced by skeletal muscle is that these stem cells are relatively rare and do not always appropriately respond to injury or disease. Indeed, as muscle undergoes normal aging processes, muscle tissue atrophies and stem cell numbers and competence appears to decline leading to much of the loss of muscle mass typical of old age in humans. Rosenthal and colleagues have identified at least one growth factor, IGF, which can restore competence to stem cells and promote myotube hypertrophy and robust appearance-mice carrying a transgene for IGF appear to resist muscle aging without collateral ill effects. The potential for application to clinical situations is now within reach. Curiously, skeletal muscle tissue is not highly prone to cancers perhaps reflecting the myogenic program which entails terminal differentiation. The field of molecular cancer cell biology has focused heavily on the role of a critical tumor suppressor gene p53 and the retinoblastoma gene, pRb. One skeletal muscle tumor which is seen primarily in children, rhabdomyosarcomas, appears to result from a novel set of molecular pathways that involve these tumor suppressors but in unexpected ways, Thayer and colleagues outline how these pathways may operate in muscle which appear to diverge greatly from the general scenario that has emerged from the study of other tumor types. vii Advances in Developmental Biology and Biochemistry Ed. Paul M. Wassarman vii -- viii © 2002 Elsevier Science. Printed in the Netherlands.
- 8. vnl When muscle differentiated in vivo, the muscle tissue expresses a variety of contractile proteins and catalytic enzymes that allow it to optimally perform its function. Specific muscles in the leg for instance, contain fast fibers which allow us to run with the drawback that they fatigue easily,. In contrast, back muscles are primarily slow which do not fatigue easily and allow us to maintain body posture for prolonged periods of time. While the contribution of nerve has long been recognized to regulate this process, it is also known that the initial pattern of fiber type specification can occur in the absence of nerve and may also reflect specific myogenic cell origin or lineages. Again, this issue has been under investigation for many years, but only recently have the signaling pathways involved become better understood. Schiaffino and colleagues outline recent progress in this field and their own work showing which pathways can regulate fiber type determination. While muscle cells can proliferate and differentiate in vitro, this process is largely manipulated by the amount of serum in the culture media. The serum which contains a mixture of growth factors or cytokines acts via specific receptors and ultimately engages components of the cell cycle and myogenic regulatory program to execute the biological outcome. It is obvious that changes in serum levels is an artificial construct of culture conditions that only vaguely reflects how differentiation is regulated in vivo. Whereas one can induce the majority of myogenic cells to terminally differentiate in vitro by lowering serum concentrations, we know that differentiation in vivo occurs in a non-synchronous fashion with fully differentiated myotubes immediately adjacent to highly proliferative myoblasts. The sheer number of cytokines implicated in this process is daunting, but a cohesive picture, as outlined by Olwin and colleagues is emerging with common effector genes downstream. Despite the intense focus on understanding skeletal myogenesis over the last several decades, investigators are often guided by certain dogmas and general ideas which influence how we interpret our data. We know, for instance, that muscle regeneration relies on the recruitment of satellite stern cells...or does it? We have defined a muscle stern cell as a cell which gives rise to skeletal muscle and does not contribute to other lineages. However work outlined by Cossu and colleagues has recently called these precepts into question. It appears that satellite cells can contribute to a wide variety of lineages and that other cell lineages such as cells originating from the bone marrow can participate in muscle regeneration. Again, the potential clinical application of this for stern cell based therapies is exciting and suggests a level of plasticity never imagined for this tissue. Thus, this volume focuses on these old and new directions for the skeletal muscle field and points out directions where the field may eventually progress. This volume is dedicated to the memory of Alex Mauro who first described the satellite cell and who spent innumerable hours with colleagues and convinced many graduate students, including me, that the field of skeletal myogenesis was one of the most fascinating biological questions to pursue. I am sure he would have something unexpected and provocative to say about where we have gone these last 10 years.
- 9. CHAPTER 1 THE MYOGENIC REGULATORY FACTORS CLAIRE M. PALMER and MICHAEL A. RUDNICKI Table of contents I. Introduction............................................................. 2 A. The MRFs are basic helix loop helix transcription factors..................... 2 B. MRF activity is regulated at the level of dimer formation..................... 3 C. MRFs bind E boxes in the promoters and enhancers of muscle specific genes..... 4 D. Identification of residues within the bHLH domains of the MRFs required for myogenic activity: The Myogenic Code................................ 4 E. Chromatin remodeling and the MRFs .................................... 5 F Myocyte enhancer factor-2 (MEF2) and myogenesis......................... 6 G. Evolutionary relationship of the MRFs ................................... 6 II. The role of the MRFs in embryonic myogenesis ................................ 7 A. Embryonic expression patterns of the MRFs ............................... 7 III. Mechanisms acting upstream of the MRFs .................................... 8 A, Pax3 ............................................................... 8 B. Synergistic regulation of muscle development by Pax3. Dach2, Eya2 and Sixl... 9 C. Development of the limb musculature: Role of Mox2 and Lbx 1............... 10 D. Axial tissues are required for epaxial somite myogenesis..................... 11 E. Shh and Wuts ........................................................ IV, Gene targeting experiments reveal a hierarchical relationship amongst the MRFs ..... 12 A. MyoD ............................................................ 12 B. Myf5 ............................................................. 12 C. MyoD and Myf5 are myogenic determination factors....................... 13 D. Myogenin is essential for myoblast differentiation in vivo but not in vitro ....... 13 E. MRF4 ............................................................ 16 F. Compound MRF mutant mice ......................................... 17 V. Postnatal myogenesis .................................................... 18 A. Satellite cells ....................................................... 18 VI. The role of the MRFs satellite cell function................................... 18 A. MRF expression pattern .............................................. 18 B. MyoD is required for satellite cell function ............................... 20 C. Myf5 and regeneration................................................ 20 D. Muscle stem cells ................................................... 21 E. Pax7 and satellite cells ............................................... 21 VII. Acknowledgements...................................................... 21 VIII. References............................................................. 22 1 Advances in Developmental Biology and Biochemistry Ed. David Sassoon 1 -- 32 © 2002 Elsevier Science. Printed in the Netherlands.
- 10. C. M. Palmer and M. A. Rudnicki I. In~oducfion The first hints to the existence of a dominant-acting myogenic transcription factor come from studies in which non-muscle cells were fused to myoblasts and muscle specific gene expression was activated in the non-muscle nuclei of the heterocaryons (Blau et al., 1983; Wright, 1984). This transcription factor was later identifiedby a subtractive cDNA approach and called MyoD ((Davis et al., 1987); for review (Weintraub et al., 1991)). MyoD is a basic helix loop helix transcription factor capable of inducing myogenic gene expression in non-muscle cells ((Davis et al., 1987); for review (Weintraub et al., 1991)). Myf5, Myogenin and MRF4 were later identified as members of the MyoD family of transcription factors, also called the myogenic regulator factors (MRFs). A similar capacity to induce muscle specific gene expression in non-muscle cells was demonstrated for these MRFs. Members of this family have subsequently been identified in a diverse array of organisms including: Drosophila (reviewed in (Abmayr et al., 1998)); Xenopus (Harvey, 1990; Hopwood et al., 1991; Jennings, 1992); Chicken (Fujisawa- Sehara et al., 1992), mouse (Davis et al., 1987; Wright et al., 1989) and human (Pearson-White, 1991). A. The MRFs are basic helix loop helix transcription factors The superfamily of basic helix loop helix (bHLH) transcriptional regulators has over 400 members which are active in a diverse array of developmental processes including myogenesis and neurogenesis (for review see (Atchley et al., 1997; Atchley et al., 1999; Morgenstern et al., 1999; Atchley et al., 2000; Wollenberg et al., 2000)). Based on homology within the bHLH domain, DNA binding affinity and the presence or absence of a leucine zipper motif, these proteins are classified into five subfamilies (Atchley et al., 1997). The MRFs are members of subfamily A, other members of this subfamily include Twist, dHand and E12 (Atchley et al., 1997). Subfamily A members bind the E Box sequence 5'- GAGCTG - 3' with high affinity, lack a leucine zipper motif and have a conserved configuration of amino acids at residues 5, 8, and 13 of the bHLH domain (Atchley et al., 1997). The four vertebrate MRFs share significant amino acid sequence identity (over 80%) throughoutthe basic helix loop helix (bHLH) domain (Murre et al., 1989b; Dias et al., 1994). The individual MRF bHLH domains appear to be functionallyequivalent however regions outside this domain give specific activities to individual MRFs (Asakura et al., 1993). Through the amphipathic helices of the bHLH domain, the MRFs can both homo and hetrodimerize (Ferre-D'Amare et al., 1993; Ma et al., 1994). The MRFs form transcription- ally active heterodimers with the widely expressed E proteins, a distinct group of bHLH proteins including E12/E47, ITF-2 and HEB (Murre et al., 1989b; Lassar et al., 1991). There is evidence to suggest that DNA binding may facilitate the dimerization of MyoD with E protein partners (Weintraub et al., 1990; Czernik et al., 1996; Maleki et al., 1997; Wendt et al., 1998).
- 11. The Myogenic Regulatory Factors B. MRF activity is regulated at the level of dimer formation Several proteins, for example Id, Mistl and MyoR. have been identified which either sequester E proteins from MyoD or prevent MyoD/E protein heterodimerization by binding the bHLH domain of MyoD (Benezra et al.. 1990; Lemercier et al., 1998; Lu et al., 1999). ld (inhibitors of DNA binding) proteins are helix loop helix proteins which lack a DNA binding domain (Benezra et al., 1990; Christy et al., 1991: Biggs et al.. 1992: Ellmeier et al., 1992: Deed et al.. 1993; Riechmann et al., 1994). Idshave been suggested to avidly bind E proteins sequestering them from bHLH transcription factors (Benezra et al.. 1990; Langlands et al., 1997). In addition. Idl and Id2 bind to MyoD and Myf5 strongly preventing dimer formation (Langlands et at.. 1997). Id expression is down regulated during myoblast differentiation presumably allowing MRF activity to regulate differentiation (Benezra et al., 1990). Furthermore. ectopic expression of Id in differentiating cultures of C2C12 myoblasts inhibits terminal differentiation (Jen et al.. 1992). Ids may function to prevent MRF dependent entry into the differentiation program during expansion of myogenic cells (Langlands et al., 1997). However, mice null for Idl, Id2. Id3. or Idl and Id3 do not have any overt muscle defects (Lyden et al.. 1999; Yokota et al., 1999; Rivera et al.. 2000; Yokota et al., 2000). The lack of a muscle phenotype may in part be due to functional redundancy between Id proteins: analysis of additional Id compound mutant mice may address this question. MyoR is a muscle restricted basic helix-loop-helix protein which antagonises MyoD (Lu et al., 1999). In the developing mouse, MyoR is expressed in a subset of muscle precursor cells predominantly during primary myogenesis between 10.5 and 16.5 days postcoitum (dpc). In cell culture, the expression of MyoR is down regulated during differentiation. In addition to sequestering E proteins. MyoR/E12 heterodimers bind E boxes within the promoters of muscle specific genes competing with MRF/E protein heterodimer for access (Lu et al., 1999). MyoR may function to modulate the timing af muscle-specific gene expression and delay muscle fiber maturation (Lu et al.. 1999). Activators of myogenesis include proteins that stimulate the heterodimerization of MyoD and E proteins. In solution the formation of MvoD/E12 heterodimers is not favoured over the formation of MyoD homodimers (Maleki et al., 1997). However, M0s-dependent phosphorylation of MyoD promotes MyoD-E12 heterodimerization while inhibiting the ?.VfyoDhomodimer DNA binding activity (Lenom~and et al.. 1997 ). c-Mos is a serine/threonine ldnase expressed in aduk skeletal muscle (Leibovitch et al.. 1991). c-Mos directly phospliorylates serine 237 of MyoD (Pelpel K et al., 2000). Mutation of this serine resMu,etO alanine abolishes the c-Mos positive regulation of MyoD (Pelpel K et al., 2000). Although in vitro data supports a role for c-Mos in :myogenic differentiation. homozygous c-Mos mutant mice do not have obvious skeletal muscle defects (Colledge et .al., 1394; tlashimoto et al., 1994; Lenormand et al., 1:997). Since the phoshorylation status of serine 237 was not determined in the c-Mos -/ mice, phosphorylation of this residue by yet4to-be identified kinases may compensate for the loss of c-Mos activity m muscle. In addition, the skeletal muscle defects in MyoD deficient mice are r~otovertly apparent expect during muscle regeneration (Rudnicki et al., 1992; Megeney et al., t996). If c-Mos is essential for MyoD activity in vivo c-mos -/- mice e~bit a muscle regeneration defect similar to mat of MyoD mutant animals.
- 12. 4 C. M. Palmer and M. A. Rudnicki C. MRFs bind E boxes in the promoters and enhancers of muscle specific genes Dimerization of bHLH proteins juxtaposes their basic domains forming a functional DNA binding domain (Ferre-D'Amare et al., 1993; Ma et al., 1994). In general, bHLH protein dimers bind to the consensus sequence 5'-CANNTG-3' (each basic region binds 1/2 of the consensus site), called an E box (Murre et al., 1989b). MyoD/E protein heterodimers preferentially bind the E box sequence 5'CA(G/C) (G/C)TG-3' (Blackwell et al., 1990). Highly conserved residues within the basic domain are responsible for consensus binding, In particular, a glutarnic acid residue conserved in the majority of bHLH proteins is in direct contact with the 'CA' nucleotides of the E box (reviewed ha (Robinson et al., 2000)). Residues within the basic domain in part dictate the consensus sequence specificity of different bHLH protein dimers. For example, Arg- 111 of MyoD makes contact with the 3'G of the E box. A similar contact does not occur when non-myogenic bHLH proteins bind DNA (reviewed in (Robinson et al., 2000)). The unique conformation of the MRF bHLH is likely due to the small size of Ala-114 (a residue required for myogenic activity). In non-myogenic bHLH proteins this position is occupied by a bulkier amino acid, it is this amino acid which is in contact with the G of CANNTG (Ferre-D'Amare et al., 1993; Ellenberger et al., 1994; Ma et al., 1994; Huang et al., 1998). D. Identification of residues within the bHLH domains of the MRFs required for myogenic activity: The myogenic code Mutagenesis studies have identified conserved residues within the basic region of the MRFs essential for their myogenic activity; residues Ala-ll4 and Thr-ll5 of MyoD and residues Ala-86 and Thr-87 of Myogenin (Davis et al., 1990; Brennan et al., 1991). These residues are referred to as the myogenic code. Replacement of Ala-114 of MyoD with asparagine or histidine, the residue normally found at that position in E proteins and Max respectively, alters the conformation of the basic domain such that Arg-111 does not make direct contact with the E box DNA (Huang et al., 1998). Furthermore this local conformational change prevents the unmasking of the N-terminal activation domain of MyoD upon DNA binding (Huang et al., 1998). Interestingly, myogenic specificity can be acquired by other bHLH proteins by the introduction of the myogenic code into the appropriate position (Weintraub et al., 1991; Davis et al., 1992). Similarly, the neurogenic bHLH protein MASH-1 acquires myogenic specificity by replacement of leucine-130 with lysine and the addition of one alpha helical turn without significantly altering cell free DNA binding properties (Dezan et al., 1999). Although the bHLH domains of the four MRFs are highly conserved, there is little sequence conservation outside this region, a notable exception is a cystein/histidine rich region (Tapscott et al., 1988; Weintraub et al., 1991). The transactivation domain is not conserved between individual MRFs. The transactivation domain of Myf5 is located in the C terminus while that of MyoD and MRF4 is in the N terminus (Braun et al., 1990; Weintraub et al., 1991; Mak et al., 1992). Strong transcriptional activation domains have been identified in both the N- and C-termini of myogenin (Olson, 1992). Additional functional studies are required to define more precisely the MRF transactivation domains.
- 13. The Myogenic Regulatory Factors E. Chromatin remodeling and the MRFs Activation of the myogenic differentiation program requires the alleviation of condensed chromatin structure. Both MyoD and Myf5 remodel chromatin in the regulatory regions of skeletal muscle specific genes (Gerber et al.. 1997). MyoD-dependem chromatin remodelling is mediated by two domains, a carboxy terminal domain and a cytseine/histidine rich domain located between the activation domain and the bHLH (Gerber et al., 1997). Interestingly, a number of previously identified MyoD binding partners (including COUP-TFII, N-CoR. p300 and PCAF) can modulate chromatin structure (Puri et al., 1997: Sartorelli et al., 1997; Bailey et al., 1998; Bailey et al.. 1999~ Sartorelli et al., 1999). The interaction of MyoD with binding partners that can modulate chromatin structure could provide a mechanism for targeting chromatin remodelling to muscle specific targets. The MyoD repressor COUP-TFII is an orphan nuclear receptor which recruits histone deacetylases via its interaction with N-CoR and other co-repressors (Bailey et al., 1998). It directly binds the N-terminal activation domain of MyoD. Interestingly MyoD also directly interacts with N-CoR through its bHLH domain (Bailey et al., 1999). In addition to recruiting histone deacetylases. COUP-TFII represses MyoD activity by competitively inhibiting the binding of p300 as a result this repression can be alleviated by overexpression of p300 fBailey et al., 1998). It and other orphan nuclear receptors including REV-erbc~ and RVR expression are down regulated during myogenic differentiation and inhibit MyoD-mediated transcription (see (Bailey et al.. 1998; Bailey et al., 1999)). These orphan nuclear receptors negatively regulate muscle differentiation and appear to have a role in maintaining the proliferative state of myoblasts (Dowries et aL, 1995; Muscat et al., 1995; Burke et al.. 1996: Bailey et al., 1998). Interestingly, MyoD recruits the histone acetyltransferase p300. and PCAF (Puri et al., 1997; Sartorelli et al., 1997; Lau et al., 1999). CBP/p300 and PCAF interact directly with MyoD and are required for MyoD-dependent transactivation (Yuan et al., 1996: Puri et al., 1997; Purl et ai.. 1997; Sartorelli et al.. 1997). A dominant negative p300 that is unable to bind MyoD inhibits both MyoD dependent myogenic conversion of 10T1/2 fibroblasts and transactivation by MyoD (Sartorelti et al., 1997), This is consistent with an essential role for endogenous p300 as a coactivator of MyoD. Both CBP/p300 and PCAF acetylate MyoD on two evolutionarily conserved lysine residues at the boundary of the 'bHLH domain (Sartorelli et ai.. 1999; Polesskaya et al., 2000). Acetylation of MyoD by either CBP/p300 or PCAF increases both in vitro DNA binding affinity and activity of muscle specific promoters (Sartorelli et al.. 1999: Polesskaya et al., 2000). Conservational substitutions of these lysine residues with nonacetylatable arginines inhibit MyoD activity (Sartorelli et al., 1999; Polesskaya et al., 2000). Recently an inhibi- tor of p300 histone acetylase activity, EID-1, was identified as a repressor of MyoD func- tion (MacLellan et al., 2000). EID-1 is a 187 amino acid E1A like protein preferentially expressed in adult cardiac and skeletal muscle tissue (MacLellan et al., 2000). Skeletal muscle specific transcription is inhibited when EID-1 is overexpressed and appears to be due to its ability to bind and inhibit p300's histone acetylase activity (MacLellan et at., 2000).
- 14. C. M. Palmer and M. A. Rudnicki F. Myocyte enhancer factor-2 (MEF2) and myogenesis The MEF2 family of transcription factors play an important role in skeletal, cardiac and smooth muscle development (for review see (Black et al., 1998)). Four vertebrate MEF2 genes have been identified (A to D) and appear to have arose from a common ancestor (for review see (Black et al., 1998)). MEF2 factors are members of the MADS-box family of transcriptional regulators. Members of this family share a 57 amino acid motif (MADS box)which provides minimal DNA-binding activity (Black et al., 1998). In addition to the MADS box, MEF2 factors contain a 29 amino acid MEF2 domain required for high affinity DNA binding and dimerization (Molkentin et al., 1996; Black et al., 1998). MEF2 proteins bind the consensus sequence 5' YTA (A/T)4 TAR 3' with varying degrees of affinity (Pollock et al., 1991; Yu et al., 1992; Molkentin et al., 1996). The transactivation domain of MEF2 factors is in the C-terminus and there is limited sequence homology in this region (for review see (Black et al., 1998)). In addition to it role as a MyoD co-factor, p300 also potentiates the transactivation by MEF2 family members (Sartorelli et al., 1997). In Drosophila, MEF2 expression is both necessary and sufficient to activate myogenesis (Bout et al., 1995; Lilly et al., 1995; Ranganayakulu et al., 1995; Lin et al., 1997). Both cardiac and skeletal muscle in Drosophila MEF2 mutant embryos fail to differentiate (Bout et al., 1995; Lilly et al., 1995; Ranganayakulu et al., 1995). However the myoblasts are appropriately specified and positioned suggesting MEF2 acts late in myogenesis to regulate terminal differentiation and fusion (Black et al., 1998). Interestingly MEF2 does not appear to be sufficient to activate the myogenic differentiation program in mammalian cells and is insufficient to induce myoblast differentiation in myogenin null mice (Yu et al., 1992; Hasty et al., 1993; Nabeshima et al., 1993; Molkentin et al., 1995; Ornatsky et al., 1997). MEF2 factors act synergistically with the MRFs to initiate myogenesis (Kaushal et al., 1994; Molkentin et al., 1995; Black et al., 1998). This synergy has been suggested to be mediated by direct MEF2/MRF interactions (Molkentin et al., 1995; Black et al., 1998). Because of functional redundancy between mouse MEF2 factors and cardiac insufficiencyresulting in lethality, MEF2 knockout mice have been uninformative in regards to the in vivo role of MEF2 factors in skeletal myogenesis (Lin et al., 1997; Bi et al., 1999). However, an essential role for MEF2 factors is suggested by studies in which a dominant-negative form of MEF2A inhibits MyoD dependent conversion of 10T1/2 fibroblasts (Omatsky et al., 1997; Kolodziejczyk et al., 1999). G. Evolutionary relationship of the MRFs Unlike vertebrates, invertebrates such as C. elegans, sea urchins and Drosophila have a single myogenic regulatory factor (Krause et al., 1990; Michelson et al., 1990; Venuti et al., 1991). Despite little sequence homology outside the bHLH domain, the functional conservation between vertebrate and invertebrate myogenic factors is remarkable (Zhang et al., 1999). The Drosophilia MRF (nautilus) and chicken MyoD rescues the myogenic defect observed in C. elegans with a loss of function mutations in CeMyoD (hlh-1) (Zhang et al., 1999).
- 15. The Myogenic Regulatory Factors 7 Phylogenic analysis suggests the four vertebrate MRFs arose from a single ancestral myogenic transcription factor (see Figure 1) (Atchley et al., 1994; Megeney et al., ~995). This factor initially gave rise to two subgroups by gene duplication. MyoD and Myf5 evolved from one subgroup and myogenin and MRF4 the other (Atchley et al.. 1994)o Interestingly, in some chordates a single gene encodes two differentially expressed MRFs suggesting there was a selective pressure for multiple MRFs prior to gene duplication (Meedel et al., 1997). Multiple MRFs likely act as specification and differentiation factors in multiple lineages allowing for coordinated muscle specific gene expression during embryogenesis (Meedel et al.. 1997). Myogenin (chr 1) Myf5 (chr 10) MRF4 (chr 10) MyoD (chr 7) Figure 1. The phylogenetic relationship of the four vertebrate MRF genes. The vertebrates MRFs arose from a common ancestor by gene duplication prior to the radiation of the vertebrates. II. The role of the MRFs in embryonic myogenesis A. Embryonic expression patterns of the MRFs In vertebrates, skeletal muscles with the exception of the anterior head. and extraocular muscles orginate from mesodermal precursor cells in the somites (for review see (Borycki et al.. 2000)). The paraxial head mesoderm gives rise to the anterior head and extraocular muscles (Wachtler et al.. 1992; Christ et al., 1995). Somites are epithelial spheres of paraxial mesoderm which flank the neural tube and form in a rostrocaudal progression (for review see (Christ et al., 1995)). As somites mature, they become subdivided into the ventral scelerotome, which contributes to the vertebra and fibs. and the dorsal dermomyotome (skeletal muscle, distal ribs and dorsal dermis) (for review see (Tajbakhsh el al., 2000)). The dorsomedial region of the dermamyotome gives rise
- 16. C. M. Palmer and M. A. Rudnicki to epaxial muscles (axial muscles of the back). The limb or hypaxial muscles arise from cells which migrate into the limb bud from the ventrolateral dermamyotome (for review see (Molkentin et al., 1996)). Signals from the neural tube, notochord and paraxial mesoderm are involved in regulating somitic myogenesis (Cossu et al., 1996). The four vertebrate MRFs have distinct spatiotemporal expression patterns. MRF expression occurs initially in rostral somites and progresses caudal with somite maturation. Myf5 is the first MRF expressed in the developing embryo. Using in situ hybridization, Myf5 is first detected in the dorsomedial quadrant of the most rostal somite at day 8 postcoitum (pc.) (Ott et al., 1991; Tajbakhsh et al., 1996). As development progresses, Myf5 expression spreads to the whole myotome and is subsequently downregulated as the myotome matures. Myf5 expression in the developing limb bud is detected between day 10 pc. and 12 pc. MyoD mRNA first apprears in the hypaxial somitic domain at 9.75 dpc (Sassoon et al., 1989; Faerman et al., 1995). MyoD expression is detected throughout the myotome by day 11.0 and is maintained throughout development. MyoD expression in the limb bud occurs together with Myf5 expression. Myogenin mRNA is detected approximately 12hrs after Myf5 mRNA in the rostral somites. Myogenin expression spreads caudally as development progresses and is maintained throughout fetal life (Sassoon et al., 1989; Ott et al., 1991). Its expression is downregulated postnatally. Myogenin mRNA is detected in the developing limb bud after 10.5 dpc. MRF-4 is expressed transiently between 9.0 and 11.5 dpc in the myotome (rostral to caudal progression). It is subsequently re-expressed in the muscles of the embryo at 16 dpc becoming the most abundant MRF mRNA postnatally (Bober et al., 1991; Hinterberger et al., 1991; Hannon et al., 1992). Successive waves of myogenic determination and differentiation likely contribute to the complex developmental expression pattern of the MRFs. III. Mechanisms acting upstream of the MRFs A. Pax3 Members of the Pax (paired box) family of proteins are developmentally important transcription factors characterized by the presence of a paired domain (Reviewed in (Borycki et al., 1997; Rawts et al., 1997)). The paired domain is a 128 amino acid DNA binding motif, which is essential for Pax protein function (Treisman et al., 1991; Xu et al., 1995). In addition to the paired domain, several Pax proteins including Pax3 and Pax7 contain a second highly conserved motif, the homeodomain. Pax3 and Pax7 are highly related proteins with similar embryonic expression patterns and as a result may be in part functional redundant (Jostes et al., 1990; Goulding et al., 1991). Identification of both Pax3/FKHR and Pax7/FKHR translocations in rhabdomyosarcomas provided the initial evidence for a role of Pax3 and Pax7 in myogenesis (Galili et al., 1993; Shapiro et al., 1993; Davis et al., 1994). Pax3 expression is initially observed in the neural tube at 8 dpc (Goulding et al., 1991). By 8.5 dpc, Pax3 expression is observed in the neuroepithelium and the dermomyotome. As development progresses, Pax3 expression in the dermomyotome is further restricted. Pax3 is also expressed in myoblasts migrating to the limb buds.
- 17. The Myogenic Regulatory Factors Mice deficient in Pax3 (Splotch mice) die in utero before day 15 pc (reviewed in (Borycki et al., 1997)). The phenotypic presentation of Pax3 null mutations is suggestive of not only a role for Pax3 in neural development but also in myogenesis. The muscles derived from migratory myoblasts (including limb muscles, back muscle (latissimus dorsi), shoulder muscles and diaphragm) are not present in Splotch embryos (10 dpc) (TajbakJash el al.. 1997). The limb muscle deficiency observed in Splotch mice is partially: the result of a failure of somite cells in the lateral dermomyotome to migrate to the limb buds (Daston et al, 1996). A similar muscle phenotype is observed in c-met receptor tyrosine kinase mutant mice, consistent with the observation that c-met is a downstream target of Pax3 (Bladt et at., 1995). Although the Pax3 expressing migrating myogenic precursor cells are committed to the myogenic lineage, they do not differentiate until reaching the appropriate location within the limb. Recent work by Bendall and colleagues have provided some clues on how these lineage committed migrating cells postpone terminal differentiation (Bendall et al., 1999). Msxl. an homeodomain containing protein, sequesters Pax3 m migrating cells preventing it from activating transcription (Bendall et at.. 1999). In addition, Msxl represses MyoD transcription by directly binding a MyoD enhancer (Woloshin et al., 1995). Although Msxl has been disrupted in mice, muscle defects have not been reported (Satokata et al., 1994; Houzelstein et al., 1997). Tajbakhsh et al. (1997) generated 1nice lacking Myf5 and Pax3. Unlike homozygous Pax3 or Myf5 mutant embryos, double homozygous mutant embryos do not express MyoD and as a result body skeletal muscle is not present in these embryos. :In addition. ectopic expression of Pax3 can activate MyoD expression in a subset of cell types (Epstein et al., 1995; Maroto et at., 1997: Tajbakhsh et al., 1997). For example ectopic Pax3 can induce myogenesis in neural tube cells, however cannot induce myogenesis m 10T1/2 fibroblasts and inhibits myogenic differentiation in C2C 12 myoblasts and 10T1/2 fibroblasts transfected with a MyoD expression plasmid (Epstein et al., 1995; Maroto e1 al.. 1997). Although Pax3 is normal expressed in the neural tube. its myogemc activity is suppressed by presumably a myogenic specific Pax3 inhibitor (RaMs et al., 1997; Maroto et al., 1997). The inability of Pax3 to induce myogenesis in other cell types may reflect either a lack of positive cofactors or the presence of Pax3 inhibitors these cells (Rawt.s et at.. 1997; Maroto et al., 1997), These findings are consistent with Pax3 acting as an upstream regulator of MyoD expression, with its ability to promote or inhibit myogenesis dependent on cell type, Similar to in vitro studies, the myogenic promoting activity of Pax3 is restricted in the embryo, suggesting that an inhibitor of Pax3 myogenic promoting activity is present in these tissues or cells (reviewed in (RaMs et al., 1997)). Alternatively an essential cofactor maybe absent from non-permissive tissues or cells (see Section 3.2 below). In addition, re~ent studies suggest Pax3 promotes expansion of the uncommitted somitic mesoderm by preYenting apoptosis of these cells (Borycki et al., 1999l, B. Synergistic regulati0n of muscle development by Pax3, Dach2. Eya2 and Six1 The genetic hierarchy regulating myogenesls is strikingly similar to that which regulates both insect and mammalian eye development (for review (Relaix et al., 1999)).
- 18. 10 C. M. Palmer and M. A. Rudnicki During eye development, eyeless (Drosophilia) or the vertebrate homolog Pax6 participates in a positive regulatory feedback loop with dachshund (Dach) and eyes absent (Eya). Both dachshund and eyes absent synergize with the homeodomain containing protein sine oculis (So) to regulate gene expression (for review (Relaix et al.. 1999; Kawakami et al., 2000)). Ectopic expression of eyeless, eyes absent or dachshund results in ectopic eye development (for review (Relaix et al., 1999; Kawakami et al., 2000)). Heanue et al. (1999) recently demonstrated that an homologous genetic network regulates vertebrate myogenesis (Heanue et al.. 1999: Relaix et al., 1999). During myogenesis, Pax3 and Dach2 appear to participate in a positive regulatory feedback loop, similar to Pax6 and Dach during eye development (Heanue et al., 1999). Dach2 can synergizes with Eya2 to induce myogenesis. In addition, Eya2 can also synergize with Sixl (a vertebrate homolog of sine oculis) to induce myogenesis (Heanue et al., 1999). C. Development of the limb musculature: Role of Mox2 and Lbx 1 Mox2 and the highly related Moxl are homeobox genes expressed in the somitic mesoderm. Moxl expression is detected prior to somite formation in the paraxial mesoderm (Candia et al., 1992; Candia et al., 1996). The development of limb musculature is perturbed in homozygous Mox2 mutant mice (Mankoo et al., 1999). These mice have a general reduction in limb musculature and are missing specific limb muscles (Mankoo et al., 1999). Unlike the Pax3 or c-met null mice, migration of myogenic precursors is normal in Mox2 deficient animals (Mankoo et al., 1999). However Mox2 is essential for the appropriate expression of both Pax3 and Myf5 in limb bud, it is not however required for appropriate MyoD expression (Mankoo et al., 1999). The mammalian homolog of the Drosophila homeobox protein ladybird, Lbxl, is essential for the lateral migration of muscle progenitor cells into the limb (Jagla et al., 1995; Brohmann et al., 2000; Gross et al., 2000). A complete absence or severe reduction of the lateral limb muscles is observed in Lbxl null mice (Schafer et al., 1999; Brohmann et al., 2000; Gross et al., 2000). Other muscles which require contributions from migratory progenitor cells including the ventral limb muscles, tongue muscle and diaphragm muscles are normal consistent with the notion that only a subset of muscle progenitors require Lbxl for appropriate migration (Schafer et al., 1999; Brohmann et al., 2000; Gross et al., 2000). Lbxl expression is detected by in situ hybridization in the lateral somite and dermomyotome at 8 dpc. By 10 dpc, Lbxl expressing migratory cells originating from the ventrolateral lip of the dermamyotome are detected, these cells are Pax3 and c-met positive (Uchiyama et al., 2000). Analysis of Lbxl expression in Splotch mice suggest Pax3 may act as an upstream regulator of Lbxl in the ventrolateral somite (Mennerich et al., 1998). Although Pax3 is required for Lbxl expression, it is not sufficient to induce Lbxl expression, signals derived from the limb bud are also necessary (Mennerich et al., 1998).
- 19. The Myogenic Regulatory Factors 11 D. Axial tissues are required for epaxial somite myogenesis Cells within the newly formed somite are positionally determined as a result of extrinisic signals acting during later stages of somite development (Aoyama et al., 1988: Ordaht et al., 1992). Analysis of the phenotypes of notochord mutant mice and zebrafish suggest an essential role of the notochord in somite myogenesis (see Figure 3) (for review see (Borycki et al.. 2000)). These studies along with grafting and explant studies suggest a combination of signals from the neural tube, notochord and surface ectoderm are sufficient to induce expression of the MRFs and somitic myogenesis (for review see IArnold et al.. 1998: Borycki et al.. 2000)). In presomitic mesoderm explants from mouse 9:5 dpc embryos, axial structures preferentially activate myogenesis through a Myf5-dependent pathway, whereas axial structure isolated from a 10.5 dpc embryo preferentially induce MyoD expression independent of Myf5 (Tajbakhsh et al., 1998). This suggests the temporal separation of Myf5 dependent and independent pathways is the result of axial tissue maturation (Tajbakhsh et al., 1998). The positive acting factors include Sonic hedgehog, and Wnt family members (see below, for review see (Borycki et al.. 2000)). E. Shh and Wnts The notochord and floor plate secrete factors including somc hedgehog which positively regulate myotome formation (for review see (Borycki et al., 2000)). Sonic hedgehog (Shh) with it two related mammalian proteins Indian hedgehog and Desert hedgehog are small secreted signaling proteins. It is synthesized in the notochord, floorplate and zone of polarizing activity in the developing embryo (Marti et al.. 1995). Shh is required for early activation of Myf5 and MyoD in somite cells that give rise to the epaxial lineage (see Figure 3) (Borycki et al., 1999). It is however not essential for the activation of Myf5 and MyoD in the hypaxial dermo- myotome or the myogenic progenitors that form limb muscle (Borycki et al.. 1999]. The downstream effectors of Shh signaling include Ptc Ithe Shh receptor) and the Gli transcription factors (for review see (Borycki et al., 200(3)). Although analysis of the expression patterns of these proteins suggest a possible role in control of myogenesls, the involvement of molecules downstream of Shh have yet to be determined. Similarly Wnt-1. Wnt-3 and Wnt-4, which are expressed in dorsal regions of the neural tu~, can activate myogeneis m vitro (see Figure 3) (Munsterberg et al.. 19957. Analysis of Wntl +, Wnt3a4- null mice suggests Wnt signals regulate early myogenesis. in particular early Myf5 expression (Ikeya et al.. 1998). Wnt7a is expressed in the dorsal ectoderm and preferentially activates MvoD (Tajbakhsh et al.. 1998). The myogenic promoting activity of Wnt7a is inhibited by BMP4. These results highlight the role of Shh and Wnt signalling in embryonic myogenesis. In addition° these results support the hypothesis that the epaxial lineage is Myf5-dependent and distinct from the MyoD-dependent hypaxial lineage.
- 20. 12 C. M. Palmer and M. A. Rudnicki IV. Gene targeting experiments reveal a hierarchical relationship amongst the MRFs The four MRFs have similar capacities to convert 10T1/2 fibroblasts into myoblasts. In addition, the DNA binding and dimerization properties are similar amongst the MRFs (Braun et al., 1991). Cross-activation of the indigenous MRF genes by ectopically expressed MRFs complicates analysis of individual MRF function (Braun et al., 1989; Thayer et al., 1989). Targeted disruption of the MRFs was essential to define the individual roles of:these proteins in myogenesis. A. MyoD Mice deficient in MyoD are viable, fertile and do not display any overt skeletal muscle abnormalities (Rudnicki et al., 1992). MyoD deficiency does not affect sacromere ultrastructure, fast/slow fiber ratio or muscle specific gene expression. Myogenin and MRF4 expression in MyoD mutant mice is unaffected. Both heterozygous and homozygous MyoD mutant mice however show significant postnatal overexpression of Myf5; 1.8 and 3.5 fold respectively (Rudnicki et al., 1992). While the epaxial muscles in a MyoD4- embryo develop normally, development of the hypaxial musculature or migrating muscle lineage is delayed by about 3.5 days (Kablar et al., 1997; Kablar et al., 1998; Kablar et al., 1999; Kaul et al., 2000). In addition, adult MyoD deficient muscle have a severe regenerative defect (Megeney et al., 1996). The effect of MyoD deficiency on regeneration will be discussed later. B. Myf5 Similar to MyoD mutant mice, newborn Myf5-/- mice have no overt morphological defects in their skeletal muscle. Expression of the three remaining MRFs is not upregulated in Myf5 mutant mice (Braun et al., 1992). Using a Myf5 targeted nls-lacZ reporter gene, Tajbakhsh and colleagues tracked the migratory routes of Myf5 expressing muscle progenitor cells ([3-gal positive cells) in both heterozygous and homozygous mutant embryos. Interestingly in Myf5 mutant embyos, muscle progenitor cells that have activated Myf5 expression ([3 -gal positive) migrate aberrantly and fail to respond to positional cues. These ceils ultimately adopt a non-myogenic fate (Tajbakhsh et al., 1996). Epaxial myogenesis in Myf5 mutant embryos is markedly delayed and is rescued by the induction of MyoD expression at 13.5 dpc (Braun et al., 1992; Kablar et al., 1997; Kablar et al., 1998; Kablar et al., 1999; Kaul et al., 2000). Hypaxial myogenesis is normal in Myf5-/-embryos. Taken together the delay in epaxial and hypaxial myogenesis observed in Myf5 mutant and MyoD mutant mice respectively supports a multiple lineage hypothesis in which Myf5 regulates epaxial muscle formation and MyoD regulates hypaxial muscle formation (Kablar et al., 1997; Kablar et al., 1998; Kablar et al., 1999). Nevertheless, the lack of obvious skeletal muscle defect in Myf5 and MyoD mutant mice suggests that the distinct myogenic lineages can to some extent compensate for each other. This hypothesis is further supported by the finding that
- 21. The Myogenic Regulatory Factors 13 Myf5 and MyoD axe not co-expressed in muscle precursor cells but rather expressed in a mutually exclusive manner (Braun et al., 1996). Several Myf5 mutant mice have been generated since the initial targeting) It was initiNly ~ocked-out by Braun et al. (1992) (Braun: et al., t992). ~ese Myf5 mutant mice die perinatally due to a severe rib defect. A similar rib defect was observ~ when a nls-lacZ,reporter gene, together with the PGK-neo selection cassette, Was inserted into exon 1 of the Myf5 locus (Tajbakhsh et al., 1996). However subsequent targeted disruption of the Myf5 locus in which the PGK-neo-TK selection cassette was excised by Cre-recombinase do not have any skeletal defects and are both viable arid fertile (Kaul et al., 2000). C. MyoD and Myf5 are myogenic determination factors To investigate whether Myf5 and MyoD could functional compensate for one another in embryonic myogenesis, compound Myf5/MyoD mutant mice were generated (Rudnicki et al., 1993), (Kaul et al., 2000). Although born alive, Myf5/MyoD compound mutant mice are immobile and die shortly after birth. These mice are devoid of both myoblasts and skeletal myofibers. Adipose and amorphous loose connective tissue occupies the spaces normally occupied by muscle (Rudnicki et al., 1993). A sequential ablation of motor neurons from the spinal cord to the brain occurs during development in Myf5+: MyoD+ mutant embryos, highlighting the intimate connection between nervous development and skeletal myogenesis (Kablar et al., 1999), The muscle phenotype of the compound mutant mice is consistent with a model in which MyoD and Myf5 acl as primary MRFs and are required for myogenic lineage determination (see Figure 2 for summary) (Rudnicki et al., 1993). Interestingly, Myf5+/-: MyoD+ mice unlike Myf5+: MyoD+/- mice are not viable suggesting postnatal survival of MyoD mutants requires two functional Myf5 alleles (Rudnicki et al, 1993). These mice have a substantial reduction in muscle fiber number. Although these observations confirm that Myf5 and MyoD can functional substitute for each other during development, they suggest Myf5 and MyoD are not functional equivalent. Similarly myogenin targeted into the Myf5 locus can partially substitute for Myf5. Myf5myg'k~lrnyg-ki. MyoD-/ mutant mice die perinatally because of reduced skeletal muscle mass (Wang et al.. 1997). The skeletal muscle formed in Myf5myg'ki/myg-ki, MyoD+ mutant mice is normal suggesting the observed reduction in skeletal muscle mass is the result of inefficient recruitment of precursors into the myogenic lineages (Wang et al., 1997). D. Myogenin is essential for myoblast differentiation m vivo but not in vitro Two independent groups generated myogenin mutant alleles by homologous recombina- tion-mediated gene targeting (Hasty et al.. 1993; Nabeshima et al., 1993). Homozygous myogenin mutant mice die perinatally presumabl5 because of a severe defect in the diaphragm. A significant reduction in diaphragm thickness and skeletal muscle content is Observed in myogenin null mice (Hasty et al.. 1993: Nabeshima et al., 1993). Unlike the MyoD -- and Myf5 -- mice. myogenin mutant mice have major skeletal
- 22. 14 C. M. Palmer and M. A. Rudnicki Ceils in Primary Secondary Somite MRFs MRFs Myf5 ~ ~ Myogenin myogenic myoblasts precursors Muscle Dots.! S~bdoma~*l ~ MyoD ~ ~ Myogenin myogenic myoblasts MRF4 ~" precursors Ventral Subdomain Muscle Figure 2. Extracellular signals from the surrounding tissue regulate myotome formation. Positive signals including Sonic hedgehog mad Wnts from the axial structures induce expression of the primary MRFs in the myotome. MyoD expression in the ventral myotome is induced by Wnt7a secreted from the ectoderm and lateral plate. The anti-myogenic activity of BMP4 is inhibited by Noggin secreted from the dorsal medial lip muscle abnormalities (Hasty et al., 1993; Nabeshima et al., 1993). A significant reduction in skeletal muscle mass is observed throughout the body. In addition, the skeletons of mygenin -/- mice show abnormal curvature of the spine and rib cage deformities. Subsequent analysis of an hypomorphic myogeninallele in which myogenin transcription is reduced to 25% wildtype, suggests distinct myogenin threshold levels are required for myogenesis and thoracic skeletal development (Vivian et al., 1999). Muscle hypoplasia is observed in both homozygous myogenin hypomophs and myogenin (hypo/null) mice. The severity of the hypoplasia is dependent on the level of myogenin expression. The thoracic skeletal defect observed in myogenin mutant mice appears to be due to delayed rib cartilage migration (Vivian et al., 1999). Histological analysis of skeletal muscle from myogenin null neonates and embryos revealed extensive disorganization of the tissue. There is a substantial reduction in myofibre density in homozygous mutant embryos. However the number of myoblasts appear normal suggesting myogenin is not required for lineage commitment but rather terminal differentiation. Consistent with this myogenin expression is upregulated during myoblast differentiation both in culture and in vivo (Sassoon et al, 1989; Wright et al., 1989; Smith et al., 1994). The generation of myogenin / Myf5 and myogenin/MyoD mutant mice demonstrates the function of myogenin does not overlap with either MyoD or Myf5 (Rawls et al., 1995). These results together suggest myogenin is a secondary MRF acting downstream of the primary MRFs, MyoD and Myf5.
- 23. The Myogenic Regulatory Factors' 15 Figure 3. Schematicrepresentation of the hierarchial geneticrelationships of the MRFsin the epaxial and hypaxialmyotOmes.Cells fromthe dorsal and ventral subdomainsdifferentiateinto myoblastsby the expressionofMyf5and MyoDrespectively.SubsequentlybothprimaryMRFsare co-expressedin thesecells. GenetargetingexperimentsdemonstratethatMyf5orMyoDarerequiredfor myogeniclineagedetermination whilemyogeninandMRF4arenecessaryforterminaldifferentiation. The muscle defect in myogenin mutant mice is most severe in muscle arising from the ventral subdomain of the dermomyotome (hypaxial) (Nabeshima et al.. t993). The hypaxial dermomyotome also gives rise to a migratory population of myogenic precursors. Very few. if any myofibres are present in these muscles. MyoD and myogenin are the predominate MRFs expressed in this region of the dermamyotome before and after myoblast differentiation respectively (Smith et al.. 1994). This is consistent with the hypothesis that myogenin is the predominate differentiation factor of a MyoD-dependent lineage. Myofibres are present in axial, intercostal and back muscles however the majority are disorganized and lack Z lines (Nabeshima et al.. 1993). Studies by Venuti and colleagues (1995) demonstrate that these myofibres arise during primary myogenesis (Venuti et al.. 1995). Suggesting primary myogenesis is unaffected by the absence of myogenin. The levels of MRF4 transcript are markedly reduced in myogenin mutant embryos while the MyoD levels are unaffected (Hasty et al., 1993: Nabeshima et al., 1993). This reduction in MRF4 could reflect a lack of differentiated myofibres as opposed to direct regulation of MRF4 by myogenin. MRF4 can partially substitute for myogenin during embryonic myogenesis (Zhu et al.. 1997). Myogenin does not appear have an essential role in myoblast differentiation in vitro, Continuous cell lines of myoblasts isolated from myogenin mutant embryos differentiate normally in culture. In addition, myogenin -- fibroblasts convert to myotubes by ectopic
- 24. 16 C. M. Palmer and M. A. Rudnicki expression of MyoD (Nabeshima et al., 1993). However. analysis of primary MyoD4- myogenic cells suggest continuous culture of primary myoblasts effects differentiation potential. The differentiation defect observed in low-passage primary MyoD+ myoblasts is not observed in later-passage MyoD+ cultures (Sabourin et al., 1999). In fight of these results, it is important to characterize the differentiation potential of low-passage primary myogenin + myoblasts. E. MRF4 The gene encoding mouse MRF4 is linked to the Myf5 gene in a head to tail orientation. Approximately 8.5 kb of sequence separates the two loci (Miner et at., 1990). Three laboratories have generated MRF4 mutant mice (Braun et at., 1995; Patapoutian et aL, 1995; Zhang et al., 1995). Although the identical PGK-neo selection cassettes was used in the targeting constructs, orientation of the cassette and deleted MRF4 sequence is unique to each construct [for review see (Olson et at., 1996)]. Analysis of the role of MRF4 in embryonic myogenesis has been complicated by the close proximity of the two MRF genes. Targeted disruption of MRF4 affects Myf5 expression via a cis-acting mechanism tFloss et at., 1996; Yoon et al., 1997). The negative cis effect of MRF4 mutations on Myf5 transcription was confirmed by the generation of compound heterozygous MRF4; Myf5 rmce in which the disrupted alleles are on different chromosomes. These mice have a significant reduction in Myf5 transcript compared to control animals suggesting the wildtype Myf5 allele is partially inactivated (Floss et at., 1996; Olson et at., 1996: Yoon et at., 1997). Myf5 enhancer elements have been identified in a 14 kb region spanning the MRF4/Myf5 locus (Summerbell et al., 2000). These enhancers regulate Myf5 expression in epaxial muscle precursor cells, the branchial arches and the central nervous system. MRF4 null mice generated in Barbara Wold's laboratory die perinatally and show defects in axial myogenesis and rib structure (Patapoutian et at., 1995). In light of recent insights into the neo cassette-dependent rib defects observed in some Myf5 mutant mice, the rib anomalies observed in the MRF4 null mice may be the result of cis effects on a yet to be identified gene (Kaul et al., 2000). A deficit in myotome development is observed in the MRF4-/- mice between 9 and 11 dpc. This corresponds to the first wave of MRF4 expression during development. A significant reduction in Myf5, MyoD and myogenin expression is observed in mutant mice at day 10 pc. A decrease in Myf5, MyoD and myogenin levels is also observed in heterozygous mice at this time (Patapoutian et al., 1995). By 14 pc, myogenesis in these MRF4 mutant animals appears normal. Minor defects are observed in the intercostal muscle of neonates (Patapoutian et al., 1995). Although studies using transgenic mice have shown the MRF4 sequences deleted in the Wold mice are not required for appropriate Myf5 expression, interference of Myf5 expression by PGK-neo can not be ruled out (Summerbell et al., 2000). The Olson MRF4 mutant mice are viable however have malformations of the ribs including bifurcations, fusions and supernumerary processes (Zhang et al., 1995). Neonatal muscle is grossly normal although the level of embryonicmyosin heavy chain is slightly decreased. Similar to the Wold MRF4-/-mice, defects in myotome differentia- tion are observed in 10.5 dpc MRF4-/- embryos (Vivian et at., 2000). Myogenin and
- 25. The Myogenic Regulatory Factors 17 Myf5 expression in the developing myotome are severely reduced indicative of retarded myotome differentiation. There is a significant increase in postnatal myogenin levels in MRF4 mutant mice. This is consistent with the notion that MRF¢ is required to down- regulate postnatal myogenin expression (Zhang et al.. 1995). Alternatively, increased myogenin expression may compensate for the loss of MRF4 (Zhang et al.. I995). The sequence deleted in the Olson MRF4 allele is more extensive than either the Arnold or Wold alleles (for review see (Olson et al.. 1996)). In addition, the PGK~neo cassette ~s inserted in the opposite orientation (opposite to Myf5 transcription), The enhancer element that regulates Myf5 epaxial somite expression is deleted in the Olson al!ele (Summerbell et al.. 2000). The reduction in Myf5 expression :in the epaxial somite may contribute to the delay in myotome differentiation observed in these MRF4 nnll mice. MRF4 mutant mice generated in the laboratory of Hans Arnold die perinatally of respiratory distress due to severe malformation of the ribs (Vivian et al., 2000). Myf5 expression is disrupted in this mutant possibly because of interference by the PGK-neo cassette (Braun et al., 1995: Summerbell et al., 2000). As a result, these mice are phenotypic representatives of Mvf5+: MRF4+ double knock-out mice. These mice are essentially a phenocopy of the Myf5 mutant mice with a delay in early myotome formation (Braun et al., 1995). The deep axial muscles are reduced in the Arnold MRF4 hornozygous knock-out mouse. Consistent with the hypothesis that Myf5 and MRF4 are important in regulating the development of muscles arising from the myotome (Braun et al., 1995). F. Compound MRF mutant mice Compound MyoD-/-:MRF44-tOlson allele) and MyoD+:Myogenin-/- mice have been generated (Rawls e~ at.. 1998). The muscle phenotype of a MRF4/ myogenin double mutant mouse is no more severe than the myogenin single mutant (Rawls et al., 1998). The number of residual myofibres is comparable in mice lacking myogenin or myogenin/MRF4 suggesting neither myogenin nor MRF4 is required for primary myofibre differentiation. Interestingly continuous lines of myoblasts isolated from MRF4/myogenin double mutant mice differentiate normally in vitro, highlighting the differences between in vivo and in vitro myogenesis. The MRF4-/-:MyoD-/- mouse is a phenocopy of the myogenin mutant mouse with only residual myofibres and undifferientated myoblasts (Rawls et al.. 1998). This is consistent with the notion that terminal differentiation of myobtasts originating from the MyoD~dependent hypaxia~ lineage requires myogenm while terminal differentiation of myoblasts originating fi'om the Myf5-dependent epaxial lineage requires MRF4. Myoblasts isolated from MyoD/MRF4 double mutant neonates differentiate normally in vitro suggesting the in vivo cellular environment may restrict the compensatory capacity of the remaining MRFs {Valdez et al.0 2000). Mice lacking all of the MRFs except Myf5 have been generated (Valdez et al.. 2000). Triple mutant myoblast fail to fuse both in vitro and in vivo. Unlike the myogenin or MRF4/MyoD mutant mice, these mice do not have residual myofibres. The observed differentiation defect in the triple mutant mice was suggested to reflect a failure to meet sufficient threshold levels of myogenic bHLH factors (Valdez et al.. 2000), (Rawls et al.. 1998). However. these results are consistent
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- 27. matters occurs in the treasury roll of 1201, where two men in Devon are set down as owing a fine “because they had been with Count John”[517]—that is, because they had supported, in his rebellion against Richard in 1193, the very man for whom, as king, the fine was now claimed. The Crown had, however, no direct means of enforcing payment of either fines or taxes, at any rate in the case of the barons. Its one remedy was to seize the lands or castles of an obstinate and wilful defaulter; and this remedy was fraught with danger to the crown itself. Neither law nor custom defined the circumstances or fixed the limits of time within which a defaulter was not, and beyond which he was, liable to be treated as obstinate and wilful; in every case where the king exercised his right of seizure on this ground, therefore, the defaulter and his friends could always find a plea for denouncing its exercise as arbitrary and unjust. It seems probable that at the close of Richard’s reign his ministers may have thus seized the castles or lands of certain barons in pledge for the arrears of their dues to the crown, and that this may have been one of the grievances referred to in the demand of the barons that Richard’s successor “should restore to each of them his rights.” John’s demand for the castles of some of the barons in 1201 was in all likelihood a proceeding of the same kind, based on the same ground, and, as it seems, equally ineffectual in compelling payment; all that the king obtained was the surrender not indeed of the castles, but of some of the barons’ sons as hostages. The deadlock was probably inevitable; but every year of its continuance aggravated both the financial difficulties of the government, and the unfriendliness of the relations between the barons and the king; and this latter evil was yet further aggravated by the measures which had necessarily to be taken in order to meet the former one. Plunged as he was from the very moment of his accession in a costly struggle with France, John had been forced to lay continually fresh burdens upon that very class among his subjects who already were, or considered themselves to be, overburdened by the demands of his predecessor. The “first scutage of King John” seems to have been assessed immediately after his coronation; it appears in the Pipe Roll
- 28. made up at Michaelmas 1199. In the financial year ending at Michaelmas 1201, and in every one of the five following years, there was another new scutage;[518] and these scutages were independent of the fines paid by the barons who did not accompany the king on his first return to Normandy in 1199, of the money taken from the host as a substitute for its service in 1201, of the equipment and payment of the “decimated” knights in 1205, and the fines claimed from all the tenants-in-chivalry after the dismissal of the host in the same year, as well as of the actual services which many of those who had paid the scutage rendered in the campaigns of 1202–1204 and 1206. The other taxes levied during these years were a carucage in 1200[519]and a seventh of moveables in 1204.[520] But all the while arrears went on accumulating, and year after year a budget had to be made up by devices of the most miscellaneous character. The accession of a new king could, of course, easily be made a pretext for selling confirmations of existing rights and privileges, and John availed himself of this pretext to the uttermost of his power at the earliest opportunity—that is, on his visit to England in 1201. During that time nobody in England seems to have felt secure of anything that he possessed till he had bought it of the king. Individuals of various ranks bought the sovereign’s “peace” or his “goodwill”;[521] the cities of Winchester and Southampton and the county of Hants each gave him money “that they might be lovingly treated”;[522] Wiltshire gave him twenty pounds “that it might be well treated.”[523] The citizens of York offended him by omitting to welcome him with a procession when he visited their city, and to provide quarters for his cross-bowmen; he demanded hostages for their future good behaviour, but afterwards changed his demand to a fine of a hundred pounds.[524] The sale of offices went on as of old; [525] while the sale of charters to towns, which under Richard was already becoming a remarkable item in the royal accounts, was a transaction of yet greater frequency and importance under his successor.[526] On the other hand, John’s treasury rolls contain many
- 29. 1207 notices of persons who owe the king money “which he has lent them.” These loans from the king to his barons and other subjects were probably made chiefly in the hope of securing the fidelity of the borrowers. In one way or another the speculation must have been in most cases a paying one for John. The privilege of claiming interest in hard cash for a loan was indeed reserved exclusively for the Jews, and not shared even by the king; but he could take from his debtors ample security on their lands or castles, or by means of hostages who were usually their sons or other young members of their families, and whom it was of the greater importance for him to hold in his power as his relations with the barons grew more strained year by year. In 1206 the tension had reached such a point that John did not venture to impose a scutage of the full amount—two marks on the knight’s fee—which had been usual since his father’s time, but contented himself with twenty shillings.[527] In 1207 he evidently dared not attempt to levy any fresh scutage at all. Nor was a carucage likely to prove either less unpopular or more productive; for the agricultural interest of the country was in a state of extreme depression, owing to a long succession of bad seasons; while the taxation of moveables was an expedient which seems to have found, as yet, but little favour with either the people or the government. John now put forth a suggestion which was, so far as we can see, a novelty in English finance. He “held a council in London on January 8, and there requested the bishops and abbots that they would allow parsons and others holding ecclesiastical benefices to give to the king a fixed sum from their revenues.”[528] Neither in equity nor in policy was the idea a bad one. While the military tenants and the socage tenants had each their own peculiar burden—scutage in the one case, carucage in the other—the beneficed clergy, as such, had never yet been subjected to taxation. The king might well argue that it was time for them to take their turn in making a special contribution to the financial needs of the State; and the argument was sure to meet with the approval of the laity. The prelates, however, were unwilling;
- 30. and the question was adjourned to another council, in which “an infinite multitude” of ecclesiastical and temporal magnates came together at Oxford on February 9. At this second meeting the bishops of both provinces gave it as their final answer that “the English Church could by no means submit to a demand which had never been heard of in all previous ages.”[529] The only approach to a precedent for it, indeed, had occurred in 1194, when Archbishop Geoffrey of York, eager to collect money for Richard’s ransom, had asked the canons of his cathedral chapter to give for that purpose a fourth part of their revenues for the year, with the result that they accused him of “wanting to overthrow the liberties of their church,” and shut its doors in his face.[530] Between the council in London and that at Oxford, Geoffrey and John, who had been more or less at variance ever since the latter’s accession, were formally reconciled;[531] John therefore probably counted upon Geoffrey’s support of his scheme, and he may have hoped that the suffragans of Canterbury, having no metropolitan of their own to lead them, would not venture to stand out against the northern primate and the king with the barons, for once, at his back. But what Geoffrey had himself asked of his own chapter as a special favour to Richard in a wholly exceptional emergency, he had no mind to give leave for John to claim from all the beneficed clergy of his province as a matter of right, and under entirely different circumstances. The king was prudent enough not to press his demand; but it may be doubted whether the lay barons agreed with the Waverley annalist in deeming its withdrawal a proof that he “had taken wiser counsel,” since he substituted for it a demand for a thirteenth of the moveable goods of every layman throughout the realm.[532] This they had no excuse for refusing. “All murmured, but no man dared contradict,”[533] except Geoffrey of York. He, it seems, claimed exemption for laymen holding lands of the Church, or at least of his cathedral church. His protest, however, was disregarded; whereupon he excommunicated all spoilers of the
- 31. 1208 Church in general, and of the province of York in particular, and then withdrew over sea,[534] to spend the rest of his life in exile. Thus for the next eight years the vast diocese of York was practically without a chief pastor and the province without a metropolitan, while the temporalities of the see were in the hand of the king. As for Canterbury, John had answered the Pope’s request that he would ratify the election of Stephen Langton by a flat refusal to accept as primate a man of whom he declared that he “knew nothing, save that he had dwelt much among his enemies”;[535] and when on June 17 Stephen was consecrated by Innocent,[536] the king seized the estates of the Canterbury chapter, drove the monks into exile,[537] and proclaimed that any one who acknowledged Stephen as archbishop should be accounted a public enemy.[538] In August Innocent bade the bishops of London, Ely and Worcester threaten the king, if he continued obstinate, with an interdict upon his realm, and hinted that this might be followed by a papal excommunication of John himself.[539] Negotiations went on throughout the winter, but without result,[540] and on Passion Sunday, March 23, or Monday, March 24, 1208, the interdict was proclaimed.[541] It seems that notice of the intended date of its publication was given about a week before, and that the king at first answered this notice by ordering all the property of the clergy, secular or monastic, to be confiscated on Monday, March 24; but that he immediately afterwards decided to anticipate, instead of returning, the blow, and caused the confiscation to be begun at once.[542] For him the opportunity was a golden one. The interdict enabled him to put the whole body of the clergy in a dilemma from which there was no escape. They held their property—thus he evidently argued—on condition of performing certain functions: if they ceased from those functions, their property was forfeit, just as that of a layman was forfeit if he withheld the service with which it was charged. The logical consequence in either case—from John’s point of view—was confiscation; difficult and dangerous to enforce
- 32. 1209 on a wide scale against laymen, but easy and safe when the victims were clergy. The barons made no objection to a proceeding which would fill the king’s coffers without drawing a single penny from their own; the chief justiciar himself, Geoffrey Fitz-Peter, earl of Essex, had no scruple in acting as custos for the Crown of all the Church property on his own estates, which were scattered through thirty- one counties, and also of the revenues and goods of the Templars throughout all England.[543] The spoliation was indeed effected with a brutal violence which would have been impossible had there been any strong feeling against it among the influential classes of the laity,[544] and which so far outran the intentions of the king that on April 11 he issued a proclamation ordering that any man caught doing or even speaking evil to a monk or a clerk, “contrary to our peace,” should be hanged upon the nearest oak.[545] The clergy, like the Jews, were to be ill-treated by no one save the king himself. Many of them made a compromise with their spoiler; within a very few weeks five bishops, three cathedral chapters, the prior of the Hospitallers, and the heads of fourteen important monasteries, besides sundry individual priests, undertook to farm their own benefices and other property for the king.[546] The Cistercians, asserting that the privileges of their order exempted them from interdict, ceased from performing the offices of religion for a few days only, and then resumed them as usual;[547] whereupon their possessions, which had been seized like those of the other orders, were restored to them on April 4.[548] At the same time John despatched an envoy to Rome proposing terms on which he professed himself willing to let Stephen take possession of his see; and he contrived to spin out the negotiations for six months before Innocent discovered that the terms offered were merely a device for wasting time, and that the king had never intended to fulfil them.[549] On January 12, 1209, the Pope informed the bishops of London, Ely and Worcester that he had written to John a letter of which he sent them a copy, and bade them excommunicate the king if he did not repent
- 33. 1208–09 within three months after its receipt.[550] John upon this began a fresh series of negotiations, which kept the three bishops—who had apparently gone over sea immediately after publishing the interdict— flitting to and fro between the continent and England, without any result, for nine more months. In October they finally withdrew, but without publishing the excommunication; and by the end of the year all possibility of its publication in England had vanished, for every English bishop had fled save two, Peter des Roches, bishop of Winchester, and John de Grey, bishop of Norwich, both of whom were creatures of the king; John de Grey, moreover, was now justiciar in Ireland, and the Poitevin Peter des Roches was thus left sole representative of the episcopal order in England.[551] It was John’s hour of triumph, not over the clergy alone, but over all his subjects and vassals within the four seas of Britain. The action of the Pope and the inaction of the barons had opened a way for him to make himself “King of England” in his own sense of the words. To all outward seeming his whole time, since his return from the continent, had been devoted to mere amusement and self-indulgence. He “haunted woods and streams, and greatly did he delight in the pleasure of them.”[552] When he was not thus chasing the beasts of the forest, his yet more relentless pursuit of other prey was making havoc of the domestic peace, and rousing against him the deadly hatred, of some of the greatest of his barons.[553] But their hatred was futile; they were paralyzed partly by their own mutual jealousies, which the king was continually stirring up,[554] partly by the consequence of their selfish shortsightedness with regard to his persecution of the clergy. The interdict had placed one whole estate of the realm at John’s mercy; and the laity, having failed at the critical moment to make common cause with their clerical brethren, now found themselves in their turn without a support against his tyranny. His consciousness of power broke out in the strangest freaks of wantonness; in causing the Michaelmas session of the Exchequer to be held at Northampton instead of London, “out of hatred to the Londoners”;[555] in
- 34. forbidding the capture of birds all over England;[556] in ordering that throughout the Forest districts the hedges should be fired and the ditches made by the people to protect their fields should be levelled, “so that, while men starved, the beasts might fatten upon the crops and fruits.”[557] It showed itself too in acts of graver political significance. A series of orders to the bailiffs of the coast towns for the equipment and mustering of their ships and the seizure of foreign vessels, issued in the spring and summer of 1208, indicates that John was then either meditating another expedition over sea, or, more probably, expecting an attack from thence. The muster, originally fixed for Trinity Sunday, was postponed to S. Matthew’s day,[558] and the end of the matter was that John, finding he had no immediate need for the services of the fleet, “took occasion”—no doubt on pretext of some deficiency in the contingent due from them—“to oppress the mariners of the Cinque Ports with great and heavy affliction. Some he hanged; some he killed with the sword; many were imprisoned and loaded with irons”; the rest fled into exile, and it was only by giving him fines and hostages that they appeased his wrath and bought his leave to return to their homes. [559] The barons were again required to renew their homage; the demand was made literally at the sword’s point—for John’s lavish hospitality and largesse[560] filled his court with mercenaries who were quite ready to enforce his will in such a matter—and they were compelled either to submit to it, or to give their sons and kinsmen as hostages for their fidelity.[561] The king seemed indeed, as Matthew Paris says, to be courting the hatred of every class of his subjects. [562] But hate him as much as they might, they feared him yet more than they hated him; and “burdensome” as he was “to both rich and poor,”[563] when he summoned all the free tenants throughout the realm, of whatever condition, who were above the age of twelve years, to swear fealty in person to him and his infant heir in the autumn of 1209, rich and poor alike durst not do otherwise than obey him.[564]
- 35. 1209 Aug. 7 This ceremony took place at Marlborough in September,[565] just before the final rupture of the negotiations with Langton and the bishops. A few weeks earlier John had received the submission of the king of Scots. Twice or thrice in the last two years a visit of William the Lion to the English court had been projected.[566] It took place at length in the middle of April 1209 at Bolton, whence John and William proceeded together to Norham for a conference.[567] The shelter given in Scotland to some of the bishops and other persons who fled from John’s persecution in connection with the interdict[568] supplied the English king with a pretext for demanding, once for all, security for William’s loyalty. He bade him surrender either three castles on the border or his only son as a hostage. William refused to do either.[569] John, on returning to the south, summoned his host, and in July set out to take the three castles by force. The papal excommunication was hanging over his head, and its publication was hourly expected; his troops shrank alike from his leadership and from an encounter with the Scot king, who was considered “eminent for his piety,” the champion of the Church and the favourite of Heaven, while they, being under interdict, were virtually outcasts from the Christian fold. A dexterous renewal of negotiations with Innocent and Stephen, however, staved off the excommunication and prevented the threatened desertion of the English troops;[570] and on August 4 John was at Norham[571] at the head of a great host ready to do battle with the Scots. On hearing this, William “greatly feared his attack, knowing him to be given to every kind of cruelty; so he came to meet him and offered to treat for peace; but the king of the English flew into a rage and insulted him bitterly, reproaching him with having received his (John’s) fugitives and public enemies into his realm, and lent them countenance and help against him.” At last some “friends of both realms” arranged terms which pacified John and which William dared not refuse. He sent his son, not indeed as a hostage, but to do homage to the English king “for the aforesaid castles and other lands which he held”;[572] he
- 36. 1199–1209 undertook to pay John by instalments within the next two years fifteen thousand marks “to have his goodwill”; he gave hostages for the fulfilment of this undertaking; and he surrendered his two daughters to be kept in John’s custody as his wards and married at his pleasure.[573] According to Gervase of Canterbury, one of these ladies was to be married to John’s son;[574] one of his many illegitimate sons must be meant, for though John had now two sons by his queen, the elder of them was not yet two years old, while the younger of William’s daughters was thirteen at the least.[575] All that William obtained in return for these concessions was the freedom of the port of Berwick, and leave to pull down a castle which the bishop of Durham had built over against it.[576] Of his claim upon Cumberland and Westmorland nothing further was ever heard. Two months later, Wales followed Scotland’s example. Over Wales, indeed, John’s triumph was won without the trouble even of a military demonstration on his part. The anarchy of Wales had been growing worse and worse ever since the death of Henry II. Its danger for England lay mainly in the opportunities which it afforded to any of the English barons of the border who might be treasonably inclined, for making alliances with one or other of the warring Welsh princes, and thus securing for themselves a support which might enable them to set at defiance the authority of the English crown. John himself had held the position of a border baron for ten years, as earl of Gloucester and lord of Glamorgan, and had used it for his own private ends as unscrupulously as any of his neighbours.[577] The familiarity with Welsh politics which he had thus acquired stood him in good stead when he became king. At his accession, a struggle which had been going on for two years between three rival claimants to the succession in South Wales, Griffith and Maelgwyn, sons of the late prince Rees ap Griffith, and Gwenwynwyn, son of Owen Cyveiliog, prince of Powys, had just ended in the triumph of Griffith, who, by the help of a force supplied to him by the English government, overcame both his rivals at the close of 1198. On Griffith’s death in
- 37. 1209–10 1200 Gwenwynwyn for a moment regained the ascendency in South Wales; but he found a new and formidable rival in the prince of North Wales, Llywelyn ap Jorwerth, who in a few years succeeded in reducing most of the South Welsh princes to dependence on himself. [578] Throughout these years John, amid all his political and military occupations on the continent, watched every vicissitude of the struggle in Wales, kept up constant relations with both parties, and balanced the one against the other[579] with a mingled unscrupulousness and dexterity for which even the Welshmen were scarcely a match, and which at last brought them all alike to his feet. In July 1202 Llywelyn promised to do homage to the English king as soon as the latter should return from over sea;[580] before October 15, 1204, he was betrothed to John’s illegitimate daughter Joan,[581] and in 1206 she became his wife.[582] In 1208 his rival Gwenwynwyn was in an English prison, whence he obtained his release by doing homage to John at Shrewsbury on October 8.[583] Llywelyn’s promised visit to the English court seems to have not yet taken place; but a year later, on the king’s return from the north, there befell, say the chroniclers, “what had never been heard of in times past: all the Welsh nobles”—that is, evidently, the princes of both North and South Wales—“came to him and did him homage,” not on the border, but in the heart of his own realm, at Woodstock, [584] on October 18 or 19, 1209.[585] The king’s triumph was complete. The last date which had been fixed for the publication of the papal sentence was October 6;[586] the sentence was still unpublished, and the bishops who should have published it had fled. They proclaimed it indeed in France in November;[587] but John took care that no official notification of the fact should reach England, and the sentence remained a dead letter. Its existence was known and talked of all over the country, but it was talked of with bated breath. The excommunicate king held his Christmas feast at Windsor surrounded by “all the great men of England,” who sat at his table
- 38. and held intercourse with him as usual, simply because they dared not do otherwise.[588] Of the fate in store for those who stood aloof, one terrible example sufficed. The archdeacon of Norwich quitted his place at the Exchequer table at Westminster, after warning his fellow-officers that they were perilling their souls by serving an excommunicate king. He was seized by a band of soldiers, loaded with chains, flung into prison, and there crushed to death beneath a cope of lead.[589] The whole body of the clergy, already stripped of their possessions, were now in peril of their lives. As the king was passing through one of the border counties he met some of the sheriff’s officers in charge of a prisoner with his hands tied behind him. They said the man was a robber, and had robbed and slain a priest on the highway: what, they asked, should be done with him? “Loose him and let him go” answered John, “he has slain one of my enemies!” Nor was his persecution limited to the clergy; the lay relatives and friends of Langton and of the other exiled bishops were hunted down and flung into prison, and their property seized for the king.[590] When he could plunder his Christian subjects no more, he turned upon the Jews. At the opening of 1210 all the Jews in England, of both sexes, were by his order arrested, imprisoned, and tortured to make them give up their wealth. It was said that the king wrung ten thousand marks from one Jew at Bristol by causing seven of his teeth to be torn out, one every day for a week,[591] and that the total sum transferred from the coffers of the Jews to the royal treasury amounted to sixty-six thousand marks.[592] Never before— not even in the worst days of William the Red—had England fallen so low as she now lay at the feet of John. “It was as if he alone were mighty upon earth, and he neither feared God nor regarded man.”[593] John seems in fact to have been one of the very few men of whom this latter assertion can be made with literal truth; and in this utter recklessness and ruthlessness lay the secret of his terrible strength. “There was not a man in the land who could resist his will in anything.”[594] The very few barons who had dared openly to
- 39. 1191–1200 resist it since his return from Poitou in 1206 were now all in Ireland; and it was Ireland that he set himself to subdue in 1210. John de Courcy had apparently ceased to be governor of the Irish March in 1191. The succession of governors there during the next few years is obscure; but we know that, as John’s chief ministers, they bore the same title which was borne by the chief minister of the king in England, that of justiciar.[595] Owing to the paucity and obscurity of the records it is difficult to gain any real understanding of the vicissitudes of the English dominion in Ireland during the twenty-five years which elapsed between John’s two visits to that country, and especially during the fourteen years between his first visit there and his accession to the English crown. He granted a new and important charter to the city of Dublin in 1192.[596] In 1195 the intruders— neither for the first nor for the last time—fell out among themselves: “John de Courcy and the son of Hugh de Lacy marched with an army to conquer the English of Leinster and Munster.”[597] They certainly did not succeed in wresting Leinster from William the Marshal. As for Munster, Richard de Cogan was apparently still holding his ground in Desmond; Raymond the Fat probably died in 1184 or 1185,[598] and as he had no direct heirs,[599] the share of that kingdom which had been originally allotted to Fitz-Stephen lapsed to John as overlord. [600] From the city of Cork the “English” are said to have been driven out in 1196;[601] but their expulsion was only momentary. Meanwhile they had at last begun to gain a footing in Thomond. By 1196 they had got possession of the city of Limerick; in that year or the next they lost it, but it was speedily recovered by Meiler Fitz- Henry,[602] who in 1199 or early in 1200 became chief justiciar in Ireland.[603] Limerick was put under the charge of William de Burgh, who apparently had won for himself some lands within the kingdom of Thomond, among them Ardpatrick, of which he received a grant from John in September 1199.[604]
- 40. 1198–1202 1179–1201 The last Irish Ard-Righ, Roderic O’Conor, died in 1198;[605] he had been dethroned sixteen years before, but his death was the signal for renewed strife between his sons for the possession of his kingdom of Connaught. The foreign settlers in Ireland took sides for their own interest in the struggle between the native princes; John de Courcy and the “English of Ulidia,” with the De Lacys of Meath and their followers, supported Cathal Crovderg O’Conor, while his rival, Cathal Carrach, was helped by “William Burke, with the English of Limerick.” For a moment Cathal Carrach’s party was victorious; but next year (1200) he was attacked by “Meiler and the English of Leinster,” while De Burgh changed sides and joined Cathal Crovderg. In 1201 or 1202 the united forces of Cathal Crovderg and De Burgh won a battle in which Cathal Carrach was slain. Cathal Crovderg being thus master of Connaught, De Burgh at once began to plot against his life; but the men of Connaught slaughtered the followers of the double-dyed traitor, and he himself escaped as best he could back to Limerick. [606] The “honour of Limerick”—exclusive of the city and the Ostmen’s cantred, which the king retained in his own hands, and the service due from the lands held within that honour by William de Burgh, which was also reserved to the Crown—had meanwhile been granted by John, on January 12, 1201, to William de Braose, “as King Henry gave it to his uncle, Philip de Braose.”[607] These last words define the extent of the “honour,” as corresponding (with the exceptions specified) to the “kingdom of Limerick” (Thomond) named in Henry’s grant of 1177. Philip de Braose was probably now dead. William was the son of Philip’s elder brother, another William who to the family estates of Bramber in Sussex and Barnstaple and Totnes in Devon had added, by his marriage with an heiress, the lordships of Radnor, Brecon, and Abergavenny in Wales.[608] The younger William probably succeeded to all these possessions soon after 1179.[609] Before 1189 his sister Maud was married to Griffith Ap Rees, who from 1198 to 1201 was
- 41. 1201–1204 Prince of South Wales; and throughout the last ten years of the twelfth century William was constantly concerned in the quarrels of the South Welsh princes and people.[610] His daughter Margaret had before November 19, 1200 become the wife of Walter de Lacy,[611] the lord of Meath, who was already her father’s neighbour on the Welsh border, where Ludlow formed part of the Lacy heritage; a younger daughter was married before 1210 to a son of another baron of the Welsh March, Roger Mortimer.[612] Count John of Mortain, as earl of Gloucester and lord of Glamorgan, was also for ten years a neighbour of William de Braose, and evidently made a friend of him, for in 1199 William was at the head of the party which most vigorously urged John’s claim to the crown.[613] In June 1200 he received a royal grant of “all the lands which he had acquired or might at any future time acquire from our Welsh enemies, to the increase of his barony of Radnor.”[614] As the king was at the same time in diplomatic relations with several of the “enemies” whom William was thus authorized to despoil, this grant was of doubtful value. The same may be said of the grant of Thomond; this, however, was a speculation on both sides; William covenanted to pay the king five thousand marks for it at the rate of five hundred marks a year.[615] De Braose immediately went to Ireland;[616] and in process of time he succeeded in obtaining possession of the greater part of his new fief, though the difficulties with which he had to contend were many and great. The other persons who had previously received from John grants of land in Thomond[617] no doubt resented and resisted the change in their position from tenants-in-chief of the king to under-tenants of William de Braose. It seems that they were upheld in their resistance by the justiciar, Meiler Fitz-Henry, and that John in consequence summoned Meiler to his court, suspended him from his office, and put it into commission in December 1201. In August 1202 John issued further orders for enforcing the claims of De Braose in Thomond; in September he forgave him all the debts which he owed to King
- 42. 1204–1206 Henry and King Richard; in October he granted the entire custody of the lands and castles of Glamorgan, Gwenllwg and Gower to “William de Braose, whose service we greatly approve.”[618] In the winter William was with the king in Normandy, and had the custody of the captive Arthur. This he resigned, seemingly at the end of the year,[619] and in January 1203 he was in charge of some matters connected with the fleet.[620] Meanwhile the governor of Limerick city, William de Burgh, had escaped from the vengeance of the Irish allies whom he had betrayed, only to fall under that of the English justiciar whom he had set at defiance. Meiler Fitz-Henry had been restored to his post; in 1203 he and Walter de Lacy joined with the Irish of Connaught in expelling De Burgh from Limerick,[621] and on July 8 William de Braose was appointed by the king to succeed De Burgh as constable of the city.[622] Meiler and De Burgh had already appealed against each other to the king;[623] in March 1204 a commission was appointed to hear their reciprocal complaints;[624] in September all De Burgh’s Irish estates except those in Connaught were restored to him on his promise of “standing to right in the King’s Court of Ireland.”[625] There is no record of the trial, which may have been prevented by his death, for at the end of the year or in 1205 he died;[626] and on April 3, 1206 the justiciar was ordered to take all his Munster estates into the king’s hand.[627] The reservation of De Burgh’s Connaught lands in 1204 may have been made in consequence of some negotiations which were at that moment going on between Meiler, as John’s representative, and the King of Connaught, Cathal Crovderg. Cathal, it seems, offered to cede two-thirds of Connaught to John, on condition that the remaining third should be secured to himself and his heirs for a yearly payment of one hundred marks. John was willing to accept this offer, but he insisted that the portion of land to be ceded to him should be chosen by Meiler, and bade Meiler take care that it was “the best part, and that which contained the best towns, ports, and
- 43. 1199–1205 sites for castles.”[628] Possibly this claim of John’s to choose the land for himself was refused by Cathal; the negotiations certainly came to nothing, for in December 1206 Cathal made another proposition. He would hold one-third of Connaught of King John for a hundred marks a year; out of the other two-thirds he would cede to John two cantreds, and for the remainder he would pay him a tribute of three hundred marks. John authorized Meiler to accept these terms, if he could get no better.[629] Whether the agreement was ever actually made, there is nothing to show; it was not likely to have any practical result. The invaders had evidently already gained some slight and precarious footing in eastern Connaught; but they had too much to do within their own March—as the dominions of the English crown in Ireland were called in those days[630]—to make any real progress westward for some years to come. The turbulence and lawlessness which prevailed in the Irish March reflected that of the Welsh March whence most of its original settlers had come. William de Braose and William de Burgh were far from being the only barons at feud with Meiler Fitz-Henry, either simply as a fellow-baron, or in his official capacity of representative of the king. In September 1199 John de Courcy and Walter de Lacy are mentioned in a royal writ as having acted together “for the destruction of our realm of Ireland.”[631] The reference probably is to their joint attack upon Leinster in 1195, which had been followed by the forfeiture of Lacy’s English and Welsh lands; these, however, he had regained in 1198.[632] In 1203, as has been seen, he helped Meiler to expel William de Burgh from Limerick; and in February 1204 he was appointed one of four commissioners to assist Meiler in dealing with escheats.[633] His former ally, John de Courcy, had a safe-conduct to and from the king’s court in July 1202;[634] but he evidently did not come to terms with the king; and next year the Lacys turned against him; Hugh de Lacy, Walter’s younger brother, defeated him in a battle near Down and drove him out of Ulidia.[635] In September he had another safe- conduct to go to the king and return “if he does not make peace
- 44. 1204 with us.”[636] This time it seems that he did “make peace,” but failed to fulfil its conditions. On August 31, 1204, he was summoned, on pain of forfeiture, to come to the king’s service “as he swore to come”; and Meiler was instructed, if the forfeiture should take place, to give to the two De Lacys the eight cantreds of De Courcy’s land which lay nearest to Meath.[637] De Courcy incurred the forfeiture; Meiler seemingly committed its execution to the De Lacys; they again attacked De Courcy, and drove him to take refuge in Tyrone; [638] and on May 2, 1205, King John granted Ulster to Hugh de Lacy, to hold “as John de Courcy held it on the day when Hugh defeated him.”[639] A few weeks later Hugh was belted earl of Ulster;[640] and at the end of June the triumph of the Lacys was completed by a royal order forbidding the chief justiciar to “move war against any man of the March” without the consent of Earl Hugh and his brother Walter.[641] With the colleagues thus forced upon him Meiler was soon at strife. His strife with Walter de Lacy, indeed, had recommenced already. Walter’s appointment as a commissioner of escheats in 1204 had been made in connexion with a demand which John—anxious to prepare for an attack upon France, as well as to guard against an expected French invasion of England, and scarcely daring to ask his English subjects for more money—addressed to all his vassals in Ireland, that they would furnish him with an aid.[642] They undertook to do so; on September 1 the king thanked them for their services and their promises, and desired that the latter might be fulfilled.[643] At the same time he was taking measures for the security of the March and of his own authority there; on August 31 he had ordered Meiler to build a castle at Dublin,[644] and in September he bade the citizens do every man his part in helping to fortify the city.[645] In November he decided upon taking back into his own hands the city of Limerick and its cantred, being, as he said, advised by his barons of England that this step was necessary for the security of his domains in Connaught and
- 45. 1207 March 1207 April 1207 Cork. It appears that William de Braose had called in the help of his son-in-law, the lord of Meath, for the keeping of this important border-post; the king’s orders for its surrender to the justiciar were addressed to Walter de Lacy and the bailiffs of William de Braose. [646] Walter seemingly refused to obey the order; Meiler, however, succeeded in taking possession of the city, “on account of which there arose a great war” between him and De Lacy,[647] with the result that John, to end their strife, took away the custody of Limerick from both of them, and restored it in August 1205 to William de Braose.[648] Nineteen months later Walter de Lacy’s castle of Ludlow was seized for the Crown, and Walter was bidden to come and “stand to right” in the English court.[649] By that time Meiler was at strife with William de Braose again, and also with another Marcher lord of very different character from any of those with whom he had as yet had to deal. Meiler Fitz-Henry, though loyal to the king, was evidently not quite the man for the post of chief justiciar in Ireland. He was one of the few survivors of the first band of Norman-Welsh adventurers who had taken part in the invasion under Robert Fitz- Stephen. The royal blood of England and of Wales was mingled in his veins; he was in fact, though not in law, first cousin to Henry II. [650] The two young Lacys, now so often opposed to him, were cousins of his wife, a niece of the elder Hugh de Lacy.[651] He was, however, not one of the great barons of the March; he seems to have held in chief of the Crown nothing except three cantreds in Desmond granted to him by John in October 1200;[652] his principal possession was the barony of Leix in Ossory,[653] for which he owed homage to William the Marshal as lord of Leinster. In the spring of 1207 William the Marshal asked leave of John to visit his Irish lands, which he had never yet seen. The leave was given, though unwillingly; but as William was on the point of setting out from Striguil, he was overtaken by a message from the king, bidding him either remain in England, or give his second son as a hostage.
- 46. 1207 Feb. 12 William sent the boy back with the messenger, saying that the king might have all his children as hostages if he pleased,[654] but as for himself, he was determined to go to Ireland; and next day he sailed. His coming was far from welcome to the justiciar, who till then had been without a superior in the country, and who resented alike the necessity of doing homage to the Marshal for the land which he held under him, and the probability of his own importance being overshadowed by the presence of a man whose territorial and personal weight was so much greater than his own. Meiler therefore wrote to the king urging him to recall the Marshal. John did so, but bade Meiler himself come over at the same time. The Marshal, though feeling that mischief was in prospect, obeyed the king’s summons with his usual readiness, and returned to England at Michaelmas, leaving his wife with a band of trusty followers to defend Leinster in his stead. Meiler also came, after secretly bidding his kinsmen and friends attack the Marshal’s lands as soon as he was gone, which they did the very next week. The king gave Meiler a warm welcome, but treated the Marshal with coldness and displeasure,[655] which Meiler soon found a way to increase. At the beginning of the year the justiciar had seized for the Crown some of the lands, men and goods of William de Braose.[656] His excuse for this proceeding was probably the fact that De Braose was in debt to the Crown for the ferm of the city of Limerick, and also for no less than four thousand two hundred and ninety-eight marks of the five thousand which he had in January 1201 covenanted to pay, by instalments of five hundred every year, for the grant of the honour of Limerick.[657] Meiler, however, had acted without instructions from the king; and when De Braose complained of the treatment which he had received, John declared that he “found no fault in him,” and bade Meiler restore everything that had been taken from him, unless indeed the city of Limerick was included; if that had been seized for the Crown, Meiler was to retain it till further orders.[658] The mingled feelings of the king are reflected in his letter. John had found in William de
- 47. 1207–08 Braose a useful servant and friend; he knew that he might find in him a dangerous enemy; he was therefore reluctant to take any measures which might drive William into opposition. On the other hand, William’s neglect of his pecuniary obligations to the Crown had reached such a pass that it could hardly be ignored much longer; and William was further suspected of being in secret alliance against the king, both with the Welsh and with the De Lacys.[659] Of this suspicion the king seems to have known nothing till after the middle of July, when he reappointed “our beloved and faithful William de Braose” custodian of Ludlow Castle.[660] It had, however, reached his ears by the time of Meiler’s coming to England, and Meiler turned it to account for a double purpose of his own. One day, as the king and his chief counsellors sat talking together after dinner, something was said about William the Marshal and his friendly relations with William de Braose. Meiler wrought upon the king’s jealousy of the one and his suspicions of the other, till he persuaded him to join in a plot for bringing them both to ruin. At the justiciar’s instigation John secretly despatched letters to all those of the Marshal’s followers in Ireland who held lands in England, bidding them, on pain of forfeiting these, to be at his court within a fortnight. At the same time Meiler, with the king’s licence, returned to Ireland. The Marshal asked permission to do the same; but this was refused. Meiler on his arrival found that hitherto his men had, on the whole, been worsted in their strife with those of Leinster. He now summoned the Marshal’s men to a “parliament,” at which the king’s messenger read out the secret letters. The men to whom these letters were addressed saw but too plainly what would be the result of their obedience: the Marshal’s lands would be left without defence against Meiler. They unanimously resolved to sacrifice their own English estates, disobey the king for their lord’s sake, and resist Meiler to the uttermost; and with the help of two powerful neighbours whom they called to their aid, Ralph Fitz-Payne and Hugh de Lacy, they succeeded, as one of them says, in doing to
- 48. 1208–09 Meiler as much mischief as he had thought to do to their lord.[661] The Marshal, meanwhile, was compelled to remain at court, but so discountenanced by the king that hardly any one dared to speak to him. At last, one winter day, as they rode out from Guildford,[662] John called to him: “Marshal, have you had any news from Ireland that pleases you?” “No, sire.” “I can tell you some news,” said the king, laughing; and he told him that his wife, the Countess Isabel, had been besieged in Kilkenny by Meiler, who had indeed been at length worsted and even captured by her people, but with very heavy losses on her side, three of the Marshal’s chief friends being among the slain. The story was a sheer invention of John’s; in reality he had received no news from Ireland at all. The Marshal, though perplexed and troubled, retained his outward composure; and early in the spring he himself received from Ireland a very different account of what had happened there. The justiciar had not only been captured, but had made submission to the countess and given his son as a hostage till he himself should stand to right in her husband’s court for the wrong which he had done to him as his lord. These tidings were sent at the same time to the king, who was by no means pleased with them, but characteristically changed his policy at once to meet the turn of the tide. He called the Marshal to his presence, greeted him with unusual courtesy, and asked him if he had heard anything from Ireland. “No, sire; I have no news from thence.” “Then I will tell you some good news, of which I wish you joy”—and thereupon John related the truth, which William knew already, though he had not chosen to say so. From that time forth “the king made him as good cheer as he had made him evil cheer before”; and when the Marshal soon afterwards again asked leave to go to Ireland, it was granted at once.[663] On March 7 Meiler was ordered to refrain from interfering with the lands of the Marshal, who had instructed his men to keep the peace towards Meiler in return;[664] on March 20 John informed the justiciar that “the Marshal has done our will,” and despatched to Ireland four commissioners by whose instructions Meiler was to act,
- 49. 1208 and who, if he failed to do so, were empowered to act in his stead. [665] On the 28th, a new grant of Leinster, on the terms of the original grant to Richard de Clare, was made by the king to the Marshal.[666] A month later Meath was in like manner granted afresh to Walter de Lacy;[667] and at the end of the next year, 1209, Meiler was removed from his office of justiciar, and replaced by the bishop of Norwich, John de Grey.[668] On one point, however, Meiler was justified by the king. In the spring of 1208 John made up his mind to bear with William de Braose no longer, and ordered a distraint upon his Welsh lands. William’s wife, Maud of Saint-Valery, [669] his nephew, Earl William of Ferrars, and his sister’s husband, Adam de Port, met the king at Gloucester and persuaded him to grant an interview to William himself at Hereford. William promised to pay his debts to the treasury within a certain time, pledged some of his castles for the payment, and gave three of his grandsons and four other persons as hostages.[670] Roger of Wendover relates that when the king’s officers went to fetch the hostages, Maud refused to deliver up her grandchildren to the king, “because,” said she, “he has murdered his captive nephew”; that her husband reproved her, and declared himself willing to answer according to law for anything in which he had offended the king; and that John, on hearing what Maud had said, was “greatly perturbed,” and ordered the whole family of De Braose to be arrested.[671] John himself, in a public statement attested by the chief justiciar of England and twelve other men of high position, among whom were De Braose’s own nephew and brother-in-law, asserted that shortly after the meeting at Hereford De Braose and his sons attempted to regain the pledged castles by force, and when they had failed in this attempt, attacked and burned Leominster.[672] Thereupon it seems that William was proclaimed a traitor; on September 21 John empowered Gerald of Athies to make an agreement with all who were or had been
- 50. 1209–10 homagers of William de Braose, so that they should “come to the king’s service and not return to the service of William.”[673] V. Stanford’s Geogˡ. Estabᵗ. London. London: Macmillan & Co., Ltd. De Braose was chased by the king’s officers,[674] till in the following year, 1209, he escaped, with his wife and two of their sons, from some Welsh seaport, intending to
- 51. go to Ireland. A violent storm kept them tossing on the sea for three days and three nights; at last they landed at Wicklow. William the Marshal chanced to be there; he received them kindly and sheltered them for three weeks. Then their presence was discovered by the new justiciar, Bishop John de Grey, who at once taxed the Marshal with harbouring “the king’s traitors,” and bade him give them up to justice. The Marshal refused, saying he had only received “his lord,”[675] as he was bound to do, and without knowing that De Braose had incurred the king’s displeasure; and he added that he himself would not act like a traitor towards De Braose at the justiciar’s bidding. Thereupon he sent the refugees safely on to their destination, the home of De Braose’s son-in-law, Walter de Lacy. The justiciar complained to the king, who summoned his host for an expedition to Ireland;[676] both the Marshal and the Lacys having positively refused to give up De Braose, though they offered to be answerable for his going to England to satisfy the king within a fixed time, and promised that, if he failed to do so, they would then harbour him no more. At last—seemingly in the spring of 1210—De Braose was allowed to go on these conditions back to Wales. John had apparently consented to meet him at Hereford; but when De Braose reached Hereford, “he,” says the king, “regarded us not,” but began to collect all the forces he could muster against the Crown. His nephew, the earl of Ferrars, however, managed to bring him to a meeting with the king at Pembroke. He offered a fine of forty thousand marks. “We,” says John, “told him we knew well that he was not in his own power at all, but in that of his wife, who was in Ireland; and we proposed that he should go to Ireland with us, and the matter should be settled there; but he chose rather to remain in Wales,”[677] and was suffered to do so—John being determined now to settle matters not only with Maud de Braose, but with all the barons of the Irish March, according to his own will and pleasure. At some date between June 16 and 20 John crossed from Pembroke to Crook, near Waterford. Thence he proceeded by way of Newbridge and Thomastown to Kilkenny, where he and all his host
- 52. 1210 were received and entertained for two days (June 23 and 24) by William the Marshal.[678] On June 28 the king reached Dublin; thence he led his host into Meath.[679] Walter de Lacy and the De Braoses fled, evidently into Ulster; thither John marched in pursuit of them, but before he could overtake them they had escaped over sea into Galloway.[680] Hugh de Lacy had retired into the stronghold of Carrickfergus; at the king’s approach, however, he, too, slipped away in a little boat to Scotland.[681] Carrickfergus was provisioned for a siege, but its garrison was soon frightened into surrender.[682] While John was at Carrickfergus, his “friend and cousin,” Duncan of Carrick, sent him word that he had captured Maud de Braose, one of her daughters, her eldest son, his wife and their two children; her younger son, Reginald, had escaped, and so had the Lacys. The king despatched John de Courcy (whom he had taken back into favour, and brought with him to Ireland, as likely to be a willing and useful helper against the De Lacys) to fetch the captives from Galloway. When they were brought before him, Maud offered the surrender of all her husband’s lands and a fine of forty thousand marks, which John accepted; but three days later she repudiated her agreement. [683] Taking his prisoners with him, the king turned southward again, and soon completed the subjugation of the Lacys’ territories. Most of the lesser barons fled before him as their lords had done, “fearing to fall into his hands.”[684] A week’s stay in Dublin (August 18 to 24) brought his expedition to a close.[685] It was probably during this second stay of John’s at Dublin that, as Roger of Wendover says, “there came to him there more than twenty kinglets[686] of that country, who all, terrified with a very great fear, did him homage and fealty; yet a few kinglets neglected to come, who scorned to do so, because they dwelt in impregnable places. Also he caused to be set up there English laws and customs, establishing sheriffs and other officers who should judge the people of that realm according to English laws.”[687] This latter statement of Roger’s may have given rise to
- 53. the later belief that it was John who organized the administration of the March in Ireland after the English model, by dividing the whole of the conquered territory into counties, each under its own sheriff. [688] It appears, however, that there were sheriffs in Ireland in the days of Henry II.[689] The earliest known mention of a sheriff’s district there occurs in 1205, when we hear of the “county of Waterford.”[690] Ten years later the same county is mentioned again, and also that of Cork;[691] and before the end of the century ten counties, at least, were recognized by the English government in Ireland.[692] The names of the earliest Irish counties thus known to us and the circumstances of John’s visit to Ireland in 1210 may suggest a clue to the rise and growth of the shire-system in that country. The district which forms the present county of Waterford had never been enfeoffed either by Henry II. or by John, but remained directly in the hands of the supreme ruler of the March. Of the present county Cork, the eastern half, at least, escheated together with the rest of Raymond FitzGerald’s share of the “kingdom of Cork” on his death about 1185. No notice of a new enfeoffment of any of the lands which had been his occurs till 1208, and then they were not granted as a whole; so far as we know, only a portion of them was enfeoffed, and that portion was distributed among several feoffees.[693] It seems probable that the system of county administration may have been first established in Ireland in those districts which were under the direct rule of the English Crown (or, to speak more exactly, of the “English,” or Angevin, “Lord of Ireland”), and of which the continuous extent was too great for them to be left, like the single cantreds attached to the other seaport towns, under the control of a mere military governor or constable, and that it was only by degrees introduced into the great fiefs. If this were so, the events of 1210 would furnish an excellent opportunity for its extension. Of the four great fiefs which, together with the royal domains and the lately redistributed honour of Cork, made up the “English” March in Ireland, Leinster was, when John sailed from Dublin for England at the end of August,[694] practically
- 54. the only one left. Meath, Ulster, and Limerick were all forfeit to the Crown; and the Crown kept the greater part of them for many years after. Meath was not restored to Walter de Lacy till 1215;[695] Walter’s brother, the earl of Ulster, did not return from exile till after John’s death;[696] and the honour of Limerick was never again bestowed as a whole upon a single grantee. Under these circumstances a system of administrative division into counties placed under sheriffs appointed by the king, or by the justiciar in his name, might be established without difficulty in territories where its introduction in earlier years, if ever attempted, would probably have been rendered ineffectual by the power of the great barons. The one great baron who in the autumn of 1210 still held his ground in the March—Earl William the Marshal, the lord of Leinster—had no hesitation in withstanding the king to his face in the cause of honour and justice; but he was not a man to throw obstacles in the way of the royal authority when it was exercised within the sphere of its rights and in the interest of public order. On the king’s return to Dublin William the Marshal came to the court. John at once accused him of having “harboured a traitor” in the person of William de Braose. The Marshal answered the king as he had answered the justiciar, and added that if any other man dared to utter such a charge against him, he was ready to disprove it there and then. As usual, no one would take up his challenge; nevertheless, John again required hostages and pledges for the Marshal’s fidelity, and again they were given at once.[697] Meanwhile, the sheriff of Hereford sent word that William de Braose was stirring up trouble in Wales, and urged that he should be outlawed; but the king ordered that the matter should await his own return to England. When he was about to sail, Maud de Braose offered to fine with him for forty thousand marks, and ten thousand in addition, as amends for having withdrawn from her former agreement. John accepted these terms; the fine was signed and sealed, and it was agreed that Maud, and also, it seems, the other members of her family who had been captured with her, should
- 55. remain in custody till it was paid. John carried his prisoners back with him to England, put Maud in prison at Bristol, and at her request gave an audience to her husband, who ratified the fine which she had made, but fled secretly just before the day fixed for paying the first instalment. The king asked Maud what she now proposed to do, and she answered plainly that she had no intention, and no means, of paying. Then it was ordered that “the judgement of our realm should be carried out against William,” and he was outlawed.[698] Thus far the king tells his own story, and there is no reason to doubt its truth. What he does not tell is the end of the story. He sent Maud and her son to a dungeon at Windsor, and there starved them to death.[699] FOOTNOTES: [Skip footnotes] [502] July 15–20, 1205, Itin. a. 7. [503] Gerv. Cant. vol. ii. p. 98. Cf. Rot. Pat. vol. i. p. 60, 60 b. [504] Cf. Innoc. III. Epp. l. viii. No. 161, and Gerv. Cant. l.c. [505] R. Wendover, vol. iii. p. 183. Cf. Gerv. Cant. vol. ii. p. 99. [506] Innoc. III. Epp. l. viii. No. 161. [507] R. Wendover, vol. iii. pp. 184, 185. [508] Rot. Pat. vol. i. p. 56 b. [509] R. Wendover, vol. iii. p. 185; Rot. Pat. l.c. [510] Innoc. III. Epp. l. ix. Nos. 34, 35, 36. [511] Rot. Pat. vol. i. pp. 65 b, 67. [512] Ib. p. 64. [513] M. Paris, Hist. Angl. vol. ii. p. 111; Chron. Maj. vol. ii. p. 514. Cf. W. Coventry, vol. ii. pp. 197, 198.
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