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© The Rockefeller University Press, 0021-9525/1998//537 $5.00
The Journal of Cell Biology, Volume 142, Number 2, , 1998 537-544

Articles

Bcl-2 Expression Identifies an Early Stage of Myogenesis and Promotes Clonal Expansion of Muscle Cells



Janice A. Dominov*,{ddagger}, Jonathan J. Dunn*, and Jeffrey Boone Miller*,{ddagger}

* Myogenesis Research Laboratory, Massachusetts General Hospital, Charlestown, Massachusetts 02129; and {ddagger} Program in Neuroscience, Harvard Medical School, Boston, Massachusetts 02115

We show that Bcl-2 expression in skeletal muscle cells identifies an early stage of the myogenic pathway, inhibits apoptosis, and promotes clonal expansion. Bcl-2 expression was limited to a small proportion of the mononucleate cells in muscle cell cultures, ranging from ~1–4% of neonatal and adult mouse muscle cells to ~5–15% of the cells from the C2C12 muscle cell line. In rapidly growing cultures, some of the Bcl-2–positive cells coexpressed markers of early stages of myogenesis, including desmin, MyoD, and Myf-5. In contrast, Bcl-2 was not expressed in multinucleate myotubes or in those mononucleate myoblasts that expressed markers of middle or late stages of myogenesis, such as myogenin, muscle regulatory factor 4 (MRF4), and myosin. The small subset of Bcl-2–positive C2C12 cells appeared to resist staurosporine-induced apoptosis. Furthermore, though myogenic cells from genetically Bcl-2–null mice formed myotubes normally, the muscle colonies produced by cloned Bcl-2–null cells contained only about half as many cells as the colonies produced by cells from wild-type mice. This result suggests that, during clonal expansion from a muscle progenitor cell, the number of progeny obtained is greater when Bcl-2 is expressed.

Key Words: apoptosis • Bcl-2 • myoblast • myogenin • skeletal muscle

Abbreviations used in this paper: BrdU, bromodeoxyuridine; MHC, myosin heavy chain; MRF, muscle regulatory factor; RT-PCR, reverse transcriptase PCR.

J.A. Dominov and J.J. Dunn contributed equally to this work.

Address all correspondence to Dr. Jeffrey B. Miller, Myogenesis Research Laboratory, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129. Tel.: (617) 726-5754. Fax: (617) 726-8543. E-mail: Miller{at}helix.mgh.harvard.edu


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