Vertebrate limb formation has been known to be initiated by a factor(s) secreted from the lateral plate mesoderm. In this report, we provide evidence that a member of the fibroblast growth factor (FGF) family, FGF10, emanates from the prospective limb mesoderm to serve as an endogenous initiator for limb bud formation. Fgf10 expression in the prospective limb mesenchyme precedes Fgf8 expression in the nascent apical ectoderm. Ectopic application of FGF10 to the chick embryonic flank can induce Fgf8 expression in the adjacent ectoderm, resulting in the formation of an additional complete limb. Expression of Fgf10 persists in the mesenchyme of the established limb bud and appears to interact with Fgf8 in the apical ectoderm and Sonic hedgehog in the zone of polarizing activity. These results suggest that FGF10 is a key mesenchymal factor involved in the initial budding as well as the continuous outgrowth of vertebrate limbs.
REFERENCES
Burke
A. C.
, Nelson
C. E.
, Morgan
B. A.
, Tabin
C.
(1995
) Hox genes and the evolution of vertebrate axial morphology.
Development
121
, 333
–346
Cohn
M. J.
, Izpisúa-Belmonte
J. C.
, Abud
H.
, Heath
J. K.
, Tickle
C.
(1995
) Fibroblast growth factors induce additional limb development from the flank of chick embryos.
Cell
80
, 739
–746
Cohn
M. J.
, Tickle
C.
(1996
) Limbs: a model for pattern formation within the vertebrate body plan.
Trends in Genetics
12
, 253
–257
Crossley
P. H.
, Martin
G. R.
(1995
) The mouse Fgf8 gene encodes a family of polypeptides and is expressed in regions that direct outgrowth and patterning in the developing embryo.
Development
121
, 439
–451
Crossley
P. H.
, Minowada
G.
, MacArthur
C. A.
, Martin
G. R.
(1996
) Roles for FGF8 in the induction, initiation, and maintenance of chick limb development.
Cell
84
, 127
–136
Dealy
C. N.
, Kosher
R. A.
(1996
) IGF-I and insulin in the acquisition of limb-forming ability by the embryonic lateral plate.
Dev. Biol
177
, 291
–299
Dhouailly
D.
, Kieny
M.
(1972
) The capacity of the flank somatic mesoderm of early bird embryos to participate in limb development.
Dev. Biol
28
, 162
–175
Fallon
J. F.
, Lopez
A.
, Ros
M. A.
, Savage
M. P.
, Olwin
B. B.
, Simandl
B. K.
(1994
) FGF-2: Apical ectodermal ridge growth signal for chick limb development.
Science
264
, 104
–107
Fekete
D. M.
, Cepko
C. L.
(1993
) Replication-competent retroviral vectors encoding alkaline phosphatase reveal spatial restriction of viral gene expression/transduction in the chick embryo.
Mol. Cell. Biol
13
, 2604
–2613
Frohman
M. A.
, Dush
M. K.
, Martin
G. R.
(1988
) Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer.
Proc. Natl. Acad. Sci. USA
85
, 8998
–9002
Gibson-Brown
J. J.
, Agulnik
S. I.
, Chapman
D. L.
, Alexiou
M.
, Garvey
N.
, Silver
L. M.
, Papaioannou
V. E.
(1996
) Evidence of a role for T-box genes in the evolution of limb morphogenesis and the specification of forelimb/hindlimb identity.
Mech. Dev
56
, 93
–101
Grieshammer
U.
, Minowada
G.
, Pisenti
J. M.
, Abbott
U. K.
, Martin
G. R.
(1996
) The chick limbless mutation causes abnormalities in limb bud dorsal-ventral patterning: implications for the mechanism of apical ridge formation.
Development
122
, 3851
–3861
Hamburger
V.
, Hamilton
H. L.
(1951
) A series of normal stages in the development of the chick embryo.
J. Morphol
94
, 257
–265
Heymann
S.
, Koudrova
M.
, Arnold
H. H.
, Köster
M.
, Braun
T.
(1996
) Regulation and function of SF/HGF during migration of limb muscle precursor cells in chicken.
Dev. Biol
180
, 566
–578
Hughes
S. H.
, Greenhouse
J. J.
, Petropoulos
C. J.
, Sutrave
P.
(1987
) Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors.
J. Virol
61
, 3004
–3012
Kuwana
T.
, Hashimoto
K.
, Nakanishi
A.
, Yasuda
Y.
, Tajima
A.
, Naito
M.
(1996
) Long-term culture of avian embryonic cells in vitro.
Int. J. Dev. Biol
40
, 1061
–1064
Laufer
E.
, Nelson
C. E.
, Johnson
R. L.
, Morgan
B. A.
, Tabin
C.
(1994
) Sonic hedgehog and Fgf-4 act through a signaling cascade and feedback loop to integrate growth and patterning of the developing limb bud.
Cell
79
, 993
–1003
Loomis
C. A.
, Harris
E.
, Michaud
J.
, Wurst
W.
, Hanks
M.
, Joyner
A. L.
(1996
) The mouse Engrailed-1 gene and ventral limb patterning.
Nature
382
, 360
–363
MacArthur
C. A.
, Lawshe
A.
, Xu
J.
, Santos-Ocampo
S.
, Heikinheimo
M.
, Chellaiah
A. T.
, Ornitz
D. M.
(1995
) FGF-8 isoforms activate receptor splice forms that are expressed in mesenchymal regions of mouse development.
Development
121
, 3603
–3613
Mahmood
R.
, Bresnick
J.
, Hornbruch
A.
, Mahony
C.
, Morton
N.
, Colquhoun
K.
, Martin
P.
, Lumsden
A.
, Dickson
C.
, Mason
I.
(1995
) A role for FGF-8 in the initiation and maintenance of vertebrate limb bud outgrowth.
Curr. Biol
5
, 797
–806
Mason
I. J.
, Fuller-Pace
F.
, Smith
R.
, Dickson
C.
(1994
) FGF-7 (keratinocyte growth factor) expression during mouse development suggests roles in myogenesis, forebrain regionalisation and epithelial-mesenchymal interactions.
Mech. Dev
45
, 15
–30
Niswander
L.
, Martin
G. R.
(1992
) Fgf-4 expression during gastrulation, myogenesis, limb and tooth development in the mouse.
Development
114
, 755
–768
Niswander
L.
, Tickle
C.
, Vogel
A.
, Booth
I.
, Martin
G. R.
(1993
) FGF-4 replaces the apical ectodermal ridge and directs outgrowth and patterning of the limb.
Cell
75
, 579
–587
Niswander
L.
, Jeffrey
S.
, Martin
G. R.
, Tickle
C.
(1994
) A positive feedback loop coordinates growth and patterning in the vertebrate limb.
Nature
371
, 609
–612
Nohno
T.
, Kawakami
Y.
, Ohuchi
H.
, Fujiwara
A.
, Yoshioka
H.
, Noji
S.
(1995
) Involvement of the Sonic hedgehog gene in chick feather formation.
Biochem. Biophys. Res. Commun
206
, 33
–39
Noramly
S.
, Pisenti
J.
, Abbott
U.
, Morgan
B.
(1996
) Gene expression in the limbless mutant: Polarized gene expression in the absence of Shh and an AER.
Dev. Biol
179
, 339
–346
Ohuchi
H.
, Yoshioka
H.
, Tanaka
A.
, Kawakami
Y.
, Nohno
T.
, Noji
S.
(1994
) Involvement of androgen-induced growth factor (FGF-8) gene in mouse embryogenesis and morphogenesis.
Biochem. Biophys. Res. Commun
204
, 882
–888
Ohuchi
H.
, Nakagawa
T.
, Yamauchi
M.
, Ohata
T.
, Yoshioka
H.
, Kuwana
T.
, Mima
T.
, Mikawa
T.
, Nohno
T.
, Noji
S.
(1995
) An additional limb can be induced from the flank of the chick embryo by FGF4.
Biochem. Biophys. Res. Commun
209
, 809
–816
Ohuchi
H.
, Shibusawa
M.
, Nakagawa
T.
, Ohata
T.
, Yoshioka
H.
, Hirai
Y.
, Nohno
T.
, Noji
S.
, Kondo
N.
(1997
) A chick wingless mutation causes abnormality in maintenance of Fgf8 expression in the wing apical ridge, resulting in loss of the dorsoventral boundary.
Mech. Dev
62
, 3
–13
Ornitz
D. M.
, Xu
J.
, Colvin
J. S.
, McEwen
D. G.
, MacArthur
C. A.
, Coulier
F.
, Gao
G.
, Goldfarb
M.
(1996
) Receptor specificity of the fibroblast growth factor family.
J. Biol. Chem
271
, 15292
–15297
Orr-Urtreger
A.
, Bedford
M. T.
, Burakova
T.
, Arman
E.
, Zimmer
Y.
, Yayon
A.
, Givol
D.
, Lonai
P.
(1993
) Developmental localization of the splicing alternatives of fibroblast growth factor receptor-2 (FGFR2).
Dev. Biol
158
, 475
–486
Pinot
M.
(1970
) Le rôle du mesoderme somitique dans la morphogeneseprecoce des membres de l'embryon de Poulet.
J. Embryol. Exp. Morph
23
, 109
–151
Riddle
R. D.
, Johnson
R. L.
, Laufer
E.
, Tabin
C.
(1993
) Sonic hedgehog mediates the polarizing activity of the ZPA.
Cell
75
, 1401
–1416
Riddle
R. D.
, Ensini
M.
, Nelson
C.
, Tsuchida
T.
, Jessell
T. M.
, Tabin
C.
(1995
) Induction of the LIM homeobox gene Lmx1 by WNT7a establishes dorsoventral pattern in the vertebrate limb.
Cell
83
, 631
–640
Ros
M. A.
, Lopez-Martinez
A.
, Simandl
B. K.
, Rodriguez
C.
, Izpisúa-Belmonte
J. C.
, Dahn
R.
, Fallon
J. F.
(1996
) The limb field mesoderm determines initial limb bud anteroposterior asymmetry and budding independent of sonic hedgehog or apical ectodermal gene expressions.
Development
122
, 2319
–2330
Rowe
D. A.
, Fallon
J. F.
(1981
) The effect of removing posterior apical ectodermal ridge of the chick wing and leg on pattern formation.
J. Embryol. Exp. Morph
65
, 309
–325
Sasaki
H.
, Hogan
B. L.
(1993
) Differential expression of multiple fork head related genes during gastrulation and axial pattern formation in the mouse embryo.
Development
118
, 47
–59
Saunders
J. W.
Jr (1948
) The proximo-distal sequence of origin of the parts of the chick wing and the role of the ectoderm.
J. Exp. Zool
108
, 363
–403
Saunders
J. W.
Jr., Gasseling
M. T.
(1963
) Trans-filter propagation of apical ectoderm maintenance factor in the chick embryo wing bud.
Dev. Biol
7
, 64
–78
Saunders
J. W.
Jr., Reuss
C.
(1974
) Inductive and axial properties of prospective wing-bud mesoderm in the chick embryo.
Dev. Biol
38
, 41
–50
Savage
M. P.
, Hart
C. E.
, Riley
B. B.
, Sasse
J.
, Olwin
B. B.
, Fallon
J. F.
(1993
) Distribution of FGF-2 suggests it has a role in chick limb bud growth.
Dev. Dyn
198
, 159
–170
Searls
R. L.
, Janners
M. Y.
(1971
) The initiation of limb bud outgrowth in the embryonic chick.
Dev. Biol
24
, 198
–213
Slack
J. M. W.
(1995
) Growth factor lends a hand.
Nature
374
, 217
–218
Stephens
T. D.
, Beier
R. L. W.
, Bringhurst
D. C.
, Hiatt
S. R.
, Prestridge
M.
, Pugmire
D. E.
, Willis
H. J.
(1989
) Limbness in the early chick embryo lateral plate.
Dev. Biol
133
, 1
–7
Streck
R. D.
, Wood
T. L.
, Hsu
M.
, Pintar
J. E.
(1992
) Insulin-like growth factor I and II and insulin-like growth factor binding protein-2 RNAs are expressed in adjacent tissues within rat embryonic and fetal limbs.
Dev. Biol
151
, 586
–596
Summerbell
D.
(1974
) A quantitative analysis of the effect of excision of the AER from the chick limb-bud.
J. Embryol. Exp. Morph
32
, 651
–660
Thery
C.
, Sharpe
M. J.
, Batley
S. J.
, Stern
C. D.
, Gherardi
E.
(1995
) Expression of HGF/SF, HGF1/MSP, and c-met suggests new functions during early chick development.
Develop. Genet
17
, 90
–101
Vogel
A.
, Rodriguez
C.
, Warnken
W.
, Izpisúa-Belmonte
J. C.
(1995
) Chick Lmx-1 specifies dorsal cells fate during vertebrate limb development.
Nature
378
, 716
–720
Vogel
A.
, Rodriguez
C.
, Izpisúa-Belmonte
J.
(1996
) Involvement of FGF-8 in initiation, outgrowth and patterning of the vertebrate limb.
Development
122
, 1737
–1750
Yamasaki
M.
, Miyake
A.
, Tagashira
S.
, Itoh
N.
(1996
) Structure and expression of the rat mRNA encoding a novel member of the fibroblast growth factor family.
J. Biol. Chem
271
, 15918
–15921
Yang
Y.
, Niswander
L.
(1995
) Interaction between the signaling molecules WNT7a and SHH during vertebrate limb development: Dorsal signals regulate anteroposterior patterning.
Cell
80
, 939
–947
Zwilling
E.
, Hansborough
L.
(1956
) Interaction between limb bud ectoderm and mesoderm in the chick embryo. III. Experiments with polydactylous limbs.
J. Exp. Zool
132
, 219
–239
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