During the development of the mouse embryo, desmin is one of the first muscle proteins detected in both the heart and the somites. The expression of the desmin gene differs from most other muscle genes, since it is initiated in replicating myoblasts and accumulates as the muscle differentiates. We have characterized a muscle-specific enhancer which directs the expression of desmin in vitro in the myoblasts and myotubes of C2 cells but not in non-myogenic cells. We report here on the generation and characterization of transgenic mice bearing a transgene in which the 1 kb DNA 5′ regulatory sequence of the desmin gene is linked to a reporter gene coding for Escherichia coli beta-galactosidase (Des1-nlacZ). The enhancer activity of the desmin promoter is very strong and the reporter gene expression is easily detected in tissue sections. We have demonstrated that the regulatory elements present in the transgene Des1-nlacZ are sufficient to direct muscle-specific and developmentally regulated expression of nlacZ in skeletal muscles. Endogenous desmin expression and transgene activity were found to be correlated during the development of skeletal muscles. The transgene was expressed in the committed mononucleate myoblasts as well as in the myotubes. In addition, we have shown that the desmin-derived sequences direct a highly selective expression of nlacZ in cells that leave the somites and invade the limb bud, indicating that the cells that migrate from the somites are already predetermined for myogenesis. In contrast, smooth and cardiac muscle cells were beta-galactosidase negative both during embryonic and foetal development. Interestingly, the transgene was found to be expressed in the conduction system of the heart, which exhibits many features characteristic of skeletal muscles.
REFERENCES
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
, 132
–142
Baldwin
T. J.
, Burden
S. J.
(1989
) Muscle-specific gene expression controlled by a regulatory element lacking a MyoD1-binding site.
Nature
341
, 716
–720
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
Brennan
T. J.
, Olson
E. N.
(1990
) Myogenin resides in the nucleus958and acquires high affinity for a conserved enhancer element on heterodimerization.
Genes Dev
4
, 582
–595
Brinster
R. L.
, Chen
H. Y.
, Trumbauer
M. E.
, Yagle
M. K.
, Palmiter
R. D.
(1985
) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs.
Proc. Natl. Acad. Sci. USA
82
, 4438
–4442
Buskin
J. N.
, Hauschka
S. D.
(1989
) Identification of a myocyte nuclear factor that binds to the muscle-specific enhancer of the mouse muscle creatine kinase gene.
Mol. Cell. Biol
9
, 2627
–2640
Caravatti
M.
, Minty
A.
, Robert
B.
, Montarras
D.
, Weydert
A.
, Cohen
A.
, Daubas
P.
, Buckingham
M.
(1982
) Regulation of muscle gene expression. The accumulation of messenger RNAs coding for muscle-specific proteins during myogenesis in a mouse cell line.
J. Mol. Biol
160
, 59
–76
Chevallier
A.
, Kieny
M.
, Mauger
A.
(1977
) Limb-somite relationship: origin of the limb musculature.
J. Embryol. Exp. Morph
41
, 245
–258
Christ
B.
, Jacob
H. J.
, Jacob
M.
(1977
) Experimental analysis of the origin of the wing musculature in avian embryos.
Anat. Embryol
150
, 171
–186
Couly
G. F.
, Coltey
P. M.
, Le Douarin
N. M.
(1992
) The developmental fate of the cephalic mesoderm in quail-chick chimeras.
Development
114
, 1
–15
Cusella-De-Anjelis
M. G.
, Lyons
G.
, Sonnino
C.
, De Angelis
L.
, Vivarelli
E.
, Farmer
K.
, Wright
W. E.
, Molinaro
M.
, Bouche
M.
, Buckingham
M.
, Cossu
G.
(1992
) MyoD, myogenin independent differenciation of primordial myoblasts in mouse somites.
J. Cell. Biol
116
, 1243
–1255
Dingwall
C.
, Laskey
R. A.
(1986
) Protein import into the cell nucleus.
Ann. Rev. Cell. Biol
2
, 367
–390
Donoghue
M.
, Ernst
H.
, Wentworth
B.
, Nadal-Ginard
B.
, Rosenthal
N.
(1988
) A muscle-specific enhancer is located at the 3end of the myosin light-chain 1/3 gene locus.
Genes Dev
2
, 1779
–1790
Furst
D. O.
, Osborn
M.
, Weber
K.
(1989
) Myogenesis in the mouse embryo: differential onset of expression of myogenic proteins and the involvement of titin in myofibril assembly.
J. Cell. Biol
109
, 517
–527
Geisler
N.
, Weber
K.
(1982
) The amino acid sequence of chiken muscle desmin provides a common structural model for intermediate filament proteins.
EMBO J
1
, 1649
–1656
Gorza
L.
, Sartore
S.
, Thornell
L. E.
, Schiaffino
S.
(1986
) Myosin type and fiber types in cardiac muscle. III. Nodal conduction tissue.
J. Cell Biol
102
, 1758
–1766
Gorza
L.
, Schiaffino
S.
, Vitadello
M.
(1987
) Heart conduction system: a neural crest derivative?.
Brain Res
457
, 360
–366
Gossett
L. A.
, Kelvin
D. J.
, Sternberg
E. A.
, Olson
E. N.
(1989
) A new myocyte-specific enhancer-binding factor that recognizes a conserved element associated with multiple muscle-specific genes.
Mol. Cell. Biol
9
, 5022
–5033
Gunning
P.
, Hardeman
E.
, Wade
R.
, Ponte
P.
, Bains
W.
, Blau
H. M.
, Kedes
L.
(1987
) Differential patterns of transcript accumulation during human myogenesis.
Mol. Cell. Biol
7
, 4100
–4114
Hannon
K.
, Smith
C. K.
II, Bales
K. R.
, Santerre
R. F.
(1992
) Temporal quantitative analysis of myogenic regulatory and growth factor gene expression in the developing mouse embryo.
Dev. Biol
151
, 137
–144
Hill
C. S.
, Duran
S.
, Lin
Z.
, Weber
K.
, Holtzer
H.
(1986
) Titin and myosin, but not desmin, are linked during myofibrillogenesis in postmitotic mononucleated myoblasts.
J. Cell. Biol
103
, 2185
–2196
Horlick
R. A.
, Benfield
P. A.
(1989
) The upstream muscle-specific enhancer of the rat muscle creatine kinase gene is composed of multiple elements.
Mol. Cell. Biol
9
, 2396
–2413
Jacob
M.
, Christ
B.
, Jacob
H. J.
(1979
) The migration of myogenic cells from the somites into the leg region of avian embryos.
Anat. Embryol
157
, 291
–309
Jaynes
J. B.
, Johnson
J. E.
, Buskin
J. N.
, Gartside
C. L.
, Hauschka
S. D.
(1988
) The muscle creatine kinase gene is regulated by multiple upstream elements, including a muscle-specific enhancer.
Mol. Cell. Biol
8
, 62
–70
Johnson
J. E.
, Wold
B. J.
, Hauschka
S. D.
(1989
) Muscle creatine kinase sequence elements regulating skeletal and cardiac muscle expression in transgenic mice.
Mol. Cell. Biol
9
, 3393
–3399
Jostes
B.
, Walther
C.
, Gruss
P.
(1991
) The murine paired box gene, Pax7, is expressed specifically during the development of the nervous and muscular system.
Mech. Dev
33
, 27
–38
Kalderon
D.
, Roberts
B. L.
, William.
D. R.
, Smith
A. E.
(1984
) A short amino acid sequence able to specify nuclear location.
Cell
39
, 499
–509
Kaufman
S. J.
, Foster
R. F.
(1988
) Replicating myoblasts express a muscle-specific phenotype.
Proc. Natl. Acad. Sci. USA
85
, 9606
–9610
Klasfeld
A.
, Bessereau
J. L.
, Salmon
A. M.
, Triller
A.
, Babinet
C.
, Changeux
J. P.
(1991
) An acetylcholine receptor-subunit promoter confering preferential synaptic expression in muscle of transgenic mice.
EMBO J
10
, 625
–632
Kozak
M.
(1987
) An analysis of 5-noncoding sequences from 699 vertebrate messenger RNAs.
Nucl. Acids Res
15
, 8125
–8131
Lassar
A. B.
, Buskin
J. N.
, Lockshon
D.
, Davis
L.
, Apone
S.
, Hauschka
D.
, Weintraub
H.
(1989
) MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer.
Cell
58
, 823
–831
Le Douarin
N. M.
(1984
) Cell migrations in embryos.
Cell
38
, 353
–360
Lee
K. J.
, Ross
R. S.
, Rockman
H. A.
, Harris
A.
, O'Brien
T.
, Van Bilsen
M.
, Shubeita
H. E.
, Kandolf
R.
, Brem
G.
, Price
J.
, Evans
S. M.
, Zhu
H.
, Franz
W. M.
, Chien
K. R.
(1992
) Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene.
J. Biol. Chem
267
, 15875
–15885
Lennon
V. A.
, Peterson
S.
, Shubert
D.
(1979
) Neurectoderm markers retained in phenotypical skeletal muscle cells arising from a glial cell line.
Nature
281
, 586
–588
Li
Z.
, Lilienbaum
A.
, Butler-Browne
G.
, Paulin
D.
(1989
) Human desmin coding gene: complete nucleotide sequence, characterization and regulation of expression during myogenesis and development.
Gene
78
, 243
–254
Li
Z.
, Paulin
D.
(1991
) High level desmin expression depends on a muscle-specific enhancer.
J. Biol. Chem
266
, 6562
–6570
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
Ordahl
C. P.
, Le Douarin
M.
(1992
) Two myogenic lineages within the developing somite.
Development
114
, 339
–353
Phillips
M. T.
, Kirby
M. L.
, Forbes
G.
(1987
) Analysis of cranial neural crest distribution in the developing heart using quail-chick chimeras,.
Circ. Res
60
, 27
–30
Pieper
F. R.
, Slobbe
R. L.
, Ramaekers
F. C. S.
, Cuypers
H. T.
, Bloemendal
H.
(1987
) Upstream regions of the hamster desmin and vimentin genes regulate expression during in vitro myogenesis.
EMBO J
6
, 3611
–3618
Quax
W.
, Broek
L. V. D.
, Egberts
W. V.
, Ramaekers
F.
, Bloemendal
H.
(1985
) Characterization of the hamster desmin gene: expression and formation of desmin filaments in nonmuscle cells after gene transfer.
Cell
43
, 327
–338
Rosenthal
N.
, Kornhauser
J. M.
, Donoghue
M.
, Rosen
K. M.
, Merlie
J. P.
(1989
) Myosin light chain enhancer activates muscle-specific, developmentally regulated gene expression in transgenic mice.
Proc. Natl. Acad. Sci.USA
86
, 7780
–7784
Sanes
J. R.
, Rubinstein
J. L.
, Nicolas
J. F.
(1986
) Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos.
EMBO. J
5
, 3133
–3142
Sanes
J.
, Johnson
Y.
, Kotzbauer
P.
, Mudd
J.
, Hanley
T.
, Martinou
J. C.
, Merlie
J.
(1991
) Selective expression of an acetylcholine receptor-lacZ transgene in synaptic nuclei of adult muscle fibers.
Development
113
, 1181
–1191
Sassoon
D.
, Lyons
G.
, Wright
W. E.
, Lin
V.
, Lassar
A.
, Weintraub
H.
, Buckingham
M.
(1989
) Expression of two myogenic regulatory factors myogenin and MyoD1 during mouse embryogenesis.
Nature
341
, 303
–307
Schaart
G.
, Viebahn
C.
, Langmann
W.
, Ramaekers
F.
(1989
) Desmin and titin expression in early postimplantation mouse embryos.
Development
107
, 585
–596
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
Sternberg
E.
, Spizz
G.
, Perry
W. M.
, Vizard
D.
, Weil
T.
, Olson
E. N.
(1988
) Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.
Mol. Cell. Biol
8
, 2896
–2909
Tam
P. P. L.
, Beddington
R. S.
(1987
) The formation of mesodermal tissues in the mouse embryo during gastrulation and early organogenesis.
Development
99
, 109
–126
Thornell
L. E.
, Eriksson
A.
(1981
) Filament system in the Purkinje fibers of the heart.
Am. J. Physiol
241
, 291
–305
Viragh
Sz.
, Challice
C. E.
(1977
) The development of the conduction system in the mouse embryo heart.
Dev. Biol
56
, 397
–411
Wang
Y. H. P. X.
, Wang
X. M.
, Ballivet
M.
, Schmidt
J.
(1988
) A cell type-specific enhancer drives expression of the chick muscle acetylcholine receptor-subunit gene.
Neuron
1
, 527
–534
Wang
X. M.
, Tsay
H. J.
, Schmidt
J.
(1990
) Expression of the acetylcholine receptor-subunit gene in differentiating chick muscle cells is activated by an element that contains two 16 bp copies of a segment of the -subunit enhancer.
EMBOJ
9
, 783
–790
Wilkinson
D. G.
, Bhatt
S.
, Chavrier
P.
, Bravo
R.
, Charnay
P.
(1989
) Segment-specific expression of a Zinc-finger gene in the developing nervous system of the mouse.
Nature
337
, 461
–463
Yutzey
K. E.
, Kline
R. L.
, Konieczny
S. F.
(1989
) An internal regulatory element controls troponin I gene expression.
Mol. Cell. Biol
9
, 1397
–1405
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