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© The Rockefeller University Press, 0021-9525/1997//331 $5.00
The Journal of Cell Biology, Volume 138, Number 2, , 1997 331-336

Article

Fusion Competence of Myoblasts Rendered Genetically Null for N-Cadherin in Culture



Carol A. Charlton*, William A. Mohler*, Glenn L. Radice{ddagger}, Richard O. Hynes{ddagger}, and Helen M. Blau*

* Department of Molecular Pharmacology, Stanford University School of Medicine, Stanford, California 94305; and {ddagger} Howard Hughes Medical Institute, Center for Cancer Research, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Myoblast fusion is essential to muscle tissue development yet remains poorly understood. N-cadherin, like other cell surface adhesion molecules, has been implicated by others in muscle formation based on its pattern of expression and on inhibition of myoblast aggregation and fusion by antibodies or peptide mimics. Mice rendered homozygous null for N-cadherin revealed the general importance of the molecule in early development, but did not test a role in skeletal myogenesis, since the embryos died before muscle formation. To test genetically the proposed role of N-cadherin in myoblast fusion, we successfully obtained N-cadherin null primary myoblasts in culture. Fusion of myoblasts expressing or lacking N-cadherin was found to be equivalent, both in vitro by intracistronic complementation of lacZ and in vivo by injection into the muscles of adult mice. An essential role for N-cadherin in mediating the effects of basic fibroblast growth factor was also excluded. These methods for obtaining genetically homozygous null somatic cells from adult tissues should have broad applications. Here, they demonstrate clearly that the putative fusion molecule, N-cadherin, is not essential for myoblast fusion.

Abbreviations used in this paper: bFGF, basic fibroblast growth factor; ES, embryonic stem.

This work was funded by grants from the National Institutes of Health (HD-18179, CA-59717, and AG-09521) and the Muscular Dystrophy Association of America to H.M. Blau, and from the National Heart Lung and Blood Institute (P01-HL41484), the National Cancer Institute (R01-CA17007), and the Howard Hughes Medical Institute to R.O. Hynes. W.A. Mohler was supported by National Institutes of Health Predoctoral Training Grants GM-07149, HD-07249, and GM-08412.

C.A. Charlton and W.A. Mohler contributed equally to this work.

Please address all correspondence to H.M. Blau, Department of Molecular Pharmacology, Stanford University School of Medicine, Stanford, CA 94305. Tel.: (415) 723-6209; Fax: (415) 725-2952; E-mail: hblau{at}cmgm.stanford.edu

W.A. Mohler's current address is Laboratory of Molecular Biology, University of Wisconsin, 1525 Linden Drive, Madison, WI 53706. Tel.: (608) 265-5428; Fax: (608) 262-4570.

G.L. Radice's current address is Division of Reproductive Biology, Department of Obstetrics and Gynecology, University of Pennsylvania, Philadelphia, PA 19104. Tel.: (215) 898-0164; Fax: (215) 573-5408.


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