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Journal of Histochemistry and Cytochemistry, Vol. 48, 1531-1544, November 2000, Copyright © 2000, The Histochemical Society, Inc.

ARTICLE

Myotube Formation is Delayed but not Prevented in MyoD-deficient Skeletal Muscle: Studies in Regenerating Whole Muscle Grafts of Adult Mice

Jason D. Whitea, Amelia Scaffidia, Marilyn Daviesa, John McGeachiea, Michael A. Rudnickib, and Miranda D. Groundsa
a Department of Anatomy and Human Biology, the University of Western Australia, Nedlands, Western Australia
b Institute for Molecular Biology and Biotechnology, McMaster University, Hamilton, Ontario, Canada

Correspondence to: Jason D. White, U. of Western Australia, Dept. of Anatomy and Human Biology, Nedlands, WA, Australia 6907. E-mail: jwhite@anhb.uwa.edu.au

We compared the time course of myogenic events in vivo in regenerating whole muscle grafts in MyoD(-/-) and control BALB/c adult mice using immunohistochemistry and electron microscopy. Immunohistochemistry with antibodies to desmin and myosin revealed a striking delay by about 3 days in the formation of myotubes in MyoD(-/-) autografts compared with BALB/c mice. However, myotube formation was not prevented, and autografts in both strains appeared similar by 8 days. Electron microscopy confirmed myotube formation in 8- but not 5-day MyoD(-/-) grafts. This pattern was not influenced by cross-transplantation experiments between strains examined at 5 days. Antibodies to proliferating cell nuclear antigen demonstrated an elevated level of replication by MyoD(-/-) myoblasts in autografts, and replication was sustained for about 3 days compared with controls. These data indicate that the delay in the onset of differentiation and hence fusion is related to extended proliferation of the MyoD(-/-) myoblasts. Overall, although muscle regeneration was delayed it was not impaired in MyoD(-/-) mice in this model. (J Histochem Cytochem 48:1531–1543, 2000)

Key Words: MyoD, skeletal muscle, regeneration, whole muscle graft, fusion, proliferation


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