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Molecular and Cellular Biology, April 1999, p. 3115-3124, Vol. 19, No. 4
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Maturation of the Myogenic Program Is Induced by
Postmitotic Expression of Insulin-Like Growth Factor I
Antonio
Musarò and
Nadia
Rosenthal*
Cardiovascular Research Center, Massachusetts
General Hospital
East, Charlestown, Massachusetts 02129
Received 22 September 1998/Returned for modification 1 December
1998/Accepted 29 December 1998
The molecular mechanisms underlying myogenic induction by
insulin-like growth factor I (IGF-I) are distinct from its
proliferative effects on myoblasts. To determine the postmitotic role
of IGF-I on muscle cell differentiation, we derived L6E9 muscle cell
lines carrying a stably transfected rat IGF-I gene under the control of
a myosin light chain (MLC) promoter-enhancer cassette. Expression of
MLC-IGF-I exclusively in differentiated L6E9 myotubes, which express
the embryonic form of myosin heavy chain (MyHC) and no endogenous
IGF-I, resulted in pronounced myotube hypertrophy, accompanied by
activation of the neonatal MyHC isoform. The hypertrophic myotubes
dramatically increased expression of myogenin, muscle creatine kinase,
-enolase, and IGF binding protein 5 and activated the myocyte
enhancer factor 2C gene which is normally silent in this cell line.
MLC-IGF-I induction in differentiated L6E9 cells also increased the
expression of a transiently transfected LacZ reporter driven by the
myogenin promoter, demonstrating activation of the differentiation
program at the transcriptional level. Nuclear reorganization,
accumulation of skeletal actin protein, and an increased expression of
1D integrin were also observed. Inhibition of the phosphatidyl
inositol (PI) 3-kinase intermediate in IGF-I-mediated signal
transduction confirmed that the PI 3-kinase pathway is required only at
early stages for IGF-I-mediated hypertrophy and neonatal MyHC induction
in these cells. Expression of IGF-I in postmitotic muscle may therefore
play an important role in the maturation of the myogenic program.
*
Corresponding author. Mailing address: Cardiovascular
Research Center, Massachusetts General Hospital
East, 149 13th St., 4th Floor, Charlestown, MA 02129-2060. Phone: (617) 724-9560. Fax:
(617) 724-9561. E-mail:
rosentha{at}helix.mgh.harvard.edu.
Molecular and Cellular Biology, April 1999, p. 3115-3124, Vol. 19, No. 4
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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