Chimeras were prepared by transplanting somites from 9-day post-coitum mouse embryos or somitic dermomyotomes from 10-day post-coitum mouse embryos into 2-day-old chick embryos at different axial levels. Mouse somitic cells then differentiated in ovo in dermis, cartilage and skeletal muscle as they normally do in the course of development and were able to migrate into chick host limb. To trace the behavior of somitic myogenic stem cells more closely, somites arising from mice bearing a transgene of the desmin gene linked to a reporter gene coding for Escherichia coli beta-galactosidase (lacZ) were grafted in ovo. Interestingly, the transgene was rapidly expressed in myotomal muscles derived from implants. In the limb muscle mass, positive cells were found several days after implantation. Activation of desmin nls lacZ also occurred in in vitro cultures of somite-derived cells. Our experimental method facilitates investigation of the mechanisms of mammalian development, allowing the normal fate of implanted mouse cells to be studied and providing suitable conditions for identification of descendants of genetically modified cells.

Aoyama
H.
,
Asamoto
K.
(
1988
)
Determination of somite cells: independence of cell differentiation and morphogenesis.
Development
104
,
15
28
Auda-Boucher
G.
,
Fontaine-Perus
J.
(
1994
)
Differentiation potentialities of distinct myogenic cell precursors in avian embryos.
Dev. Dyn
201
,
95
107
Babai
F.
,
Musevi-Aghdam
J.
,
Schurch
W.
,
Royal
A.
,
Gabbiani
G.
(
1990
)
Coexpression of-sarcomeric actin, -smooth muscle actin and desmin during myogenesis in rat and mouse embryos.
I. Skeletal muscle. Differentiation
44
,
1382
142
Beddington
R. S. P.
,
Martin
P.
(
1989
)
An in situ transgenic enzyme marker to monitor migration of cells in the mid-gestation mouse embryo: somite contribution to the limb.
Mol. Biol. Med
6
,
263
274
Beresford
B.
(
1983
)
Brachial muscles in the chick embryo: the fate of individual somites.
J. Embryol. Exp. Morph
77
,
99
116
Bober
E.
,
Lyons
G. E.
,
Braun
T.
,
Cossu
G.
,
Buckingham
M.
,
Arnold
H. H.
(
1991
)
The muscle regulatory gene, myf-6 has a biphasic pattern of expression during early mouse development.
J. Cell Biol
133
,
1255
1265
Bober
E.
,
Franz
T.
,
Arnold
H. H.
,
Gruss
P.
,
Tremblay
J. P.
(
1994
)
Pax3 is required for the development of limb muscles: a possible role for the migration of dermomyotomal muscle progenitor cells.
Development
120
,
603
612
Borman
W. H.
,
Yorde
D. E.
(
1994
)
Analysis of chick somite myogenesis by in situ confocal microscopy of desmin expression.
J. Histochem. Cytochem
42
,
265
272
Borman
W. H.
,
Urlakis
K. J.
,
Yorde
D. E.
(
1994
)
Analysis of the in vivo myogenesis status of chick somites by desmin expression in vitro.
Dev. Dyn
199
,
268
279
Braun
T.
,
Rudnicki
M.
,
Arnold
H.
,
Jaenisch
R.
(
1992
)
Targeted inactivation of the muscle regulatory gene myf-5 results in abnormal rib development and perinatal death.
Cell
71
,
369
382
Cheng
T. S.
,
Hanley
T. A.
,
Mudd
J.
,
Merlie
J. P.
,
Olson
E. N.
(
1992
)
Mapping of myogenin transcription during embryogenesis using transgenes linked to the myogenin control region.
J. Cell Biol
119
,
1649
1656
Chevallier
A.
(
1977
)
Origine des ceintures scapulaires et pelviennes chez l'embryon d'oiseau.
J. Embryol. Exp. Morph
42
,
275
292
Chevallier
A.
,
Kieny
M.
,
Mauger
A.
(
1977
)
Limb-somite relationship: origin of the limb musculature.
J. Embryol. Exp. Morph
41
,
245
258
Chevallier
A.
,
Kieny
M.
,
Mauger
A.
(
1978
)
Limb-somite relationship: effect of removal of somitic mesoderm on the wing musculature.
J. Embryol. Exp. Morph
43
,
263
278
Christ
B.
,
Jacob
H.
,
Jacob
M.
(
1977
)
Experimental analysis of the origin of the wing musculature in avian embryos.
Anat. Embryol
150
,
171
186
Christ
B.
,
Jacob
H.
,
Jacob
M.
(
1978
)
On the formation of the myotomes in avian embryos. An experimental and scanning electron microscope study.
Experientia
34
,
514
516
Emerson
C. P.
(
1990
)
Myogenesis and developmental control genes.
Curr. Op. Gen. Dev
2
,
1065
1075
Emerson
C. P.
(
1993
)
Skeletal myogenesis: genetics and embryology to the fore.
Curr. Op. Gen. Dev
3
,
265
274
Fontaine-Perus
J.
,
Chanconie
M.
,
Le Douarin
N. M.
(
1985
)
Embryonic origin of substance P-containing neurons in cranial and spinal sensory ganglia.
Dev. Biol
107
,
227
238
Franz
T.
(
1993
)
The Splotch (Sp1H) and Splotch-delayed (Spd) alleles: differential phenotypic effects on neural crest and limb musculature.
Anat. Embryol
187
,
371
377
Franz
T.
,
Kothary
R.
,
Surani
M. A.
,
Halata
Z.
,
Grim
M.
(
1993
)
The Splotch mutation interferes with muscle development in the limbs.
Anat. Embryol
187
,
153
160
Geisler
N.
,
Weber
K.
(
1982
)
The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins.
EMBO J
1
,
1649
1656
Goulding
M.
,
Lumsden
A.
,
Paquette
A. J.
(
1994
)
Regulation of Pax-3 expression in the dermomyotome and its role in muscle development.
Development
120
,
957
971
Gruss
P.
,
Walther
C.
(
1992
)
Pax in development.
Cell
69
,
719
722
Hamburger
V.
,
Hamilton
H. L.
(
1951
)
A series of normal stages in the development of the chick embryo.
J. Morph
88
,
49
92
Hasty
P.
,
Bradley
A.
,
Morris
J. H.
,
Edmonson
D. G.
,
Venuti
J. M.
,
Olson
E. N.
,
Klein
W. H.
(
1993
)
Muscle deficiency and neonatal death in mice with a targeted mutation in the myogenin gene.
Nature
364
,
501
506
Hinterberger
J. T.
,
Sassoon
D. A.
,
Rhodes
S. J.
,
Konieczny
S. F.
(
1991
)
Expression of the muscle regulatory factor MRF4 during somite and skeletal myofiber development.
Dev. Biol
147
,
144
156
Laframboise
W. A.
,
Daood
M. J.
,
Guthrie
R. D.
,
Schiaffino
S.
,
Moretti
L.
,
Brozanski
B.
,
Ontell
M. P.
,
Butler-Browne
G. S.
,
Whalen
R. G.
,
Ontell
M.
(
1991
)
Emergence of a mature myosin phenotype in the rat diaphragm muscle.
Dev. Biol
144
,
1
15
Lazarides
E.
,
Hubbard
B. D.
(
1976
)
Immunological characterization of the subunit of the 1OO A filaments from muscle cells.
Proc. Natl. Acad. Sci. USA
73
,
4344
4348
Lee
K. H.
,
Sze
L. Y.
(
1993
)
Role of the brachial somites in the development of appendicular musculature in rat embryos.
Dev. Dyn
198
,
86
93
Li
Z.
,
Marchand
P.
,
Babinet
C.
,
Paulin
D.
(
1993
)
Desmin sequence elements regulating skeletal muscle-specific expression in transgenic mice.
Development
117
,
947
959
Lyons
G. E.
,
Ontell
M.
,
Cox
R.
,
Sassoon
D.
,
Buckingham
M.
(
1990
)
The expression of myosin genes in developing skeletal muscle in the mouse embryo.
J. Cell Biol
111
,
1465
1476
Mayo
M.
,
Brinbas
M.
,
Santos
V.
,
Shum
L.
,
Slavkin
H. C.
(
1992
)
Desmin expression during early mouse tongue morphogenesis.
J. Dev. Biol
36
,
255
263
Moscona
A.
(
1957
)
The development in vitro of chimeric aggregates of dissociated embryonic chick and mouse cells.
Proc. Natl. Acad. Sci. USA
43
,
184
194
Nabeshima
Y.
,
Hanaoka
K.
,
Hayasaka
M.
,
Esumi
E.
,
Li
S.
,
Nonaka
I.
,
Nabeshima
Y.
(
1993
)
Myogenein gene disruption results in perinatal lethality because of severe defect.
Nature
364
,
532
535
Olson
E. N.
(
1992
)
Interplay between proliferation and differentiation within the myogenic lineage.
Dev. Biol
154
,
261
272
Ordahl
C. P.
,
Le Douarin
N.M.
(
1992
)
Two myogenic lineages within the developing somite.
Development
114
,
339
353
Ott
M.
,
Bober
E.
,
Lyons
G.
,
Arnold
H.
,
Buckingham
M.
(
1991
)
Early expression of the myogenic regulatory gene, myf-5, in precursor cells of skeletal muscle in the mouse embryo.
Development
111
,
1097
1107
Rudnicki
M. A.
,
Braun
T.
,
Hinuma
S.
,
Jaenisch
R.
(
1992
)
Inactivation of MyoD in mice leads to up-regulation of the myogenic HLHgene myogenin and results in apparently normal muscle development.
Cell
71
,
383
390
Rudnicki
M. A.
,
Schnegelsberg
P. N.
,
Stead
R. H.
,
Braun
T.
,
Arnold
H. H.
,
Jaenish
R.
(
1993
)
MyoD or Myf5 is required for the formation of skeletal muscle.
Cell
15
,
1531
1539
Sassoon
D.
,
Lyons
G.
,
Wright
W. E.
,
Lin
V.
,
Lassar
A.
,
Weintraub
H.
,
Buckingham
M.
(
1989
)
Expression of two myogenic regulatory factors myogenin and MyoD during mouse embryogenesis.
Nature
341
,
303
307
Small
J. V.
,
Sobieszek
A.
(
1977
)
Studies on the function and composition of the 10-nm (100-A) filaments of vertebrate smooth muscle.
J. Cell Sci
23
,
243
268
Tajbakhsh
S.
,
Buckingham
M.
(
1994
)
Mouse limb muscle is determined in the absence of the earliest myogenic factor Myf5.
Proc. Natl. Acad. Sci. USA
91
,
747
751
Tam
P. P. L.
(
1981
)
The control of somitogenesis in mouse embryos.
J. Embryol. Exp. Morph
65
,
103
128
Williams
B. A.
,
Ordahl
C.
(
1994
)
Pax-3 expression in segmental mesoderm marks early stages in myogenic cell specification.
Development
120
,
785
796
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