Embryonic and fetal rat myoblasts express different phenotypes following differentiation in vitro

Abstract

Myosin heavy chain (MHC) is encoded by a multigene family containing members which are expressed in developmental and fiber type-specific patterns. In developing rats, primary (1 degree) and secondary (2 degrees) myotubes can be distinguished by difference in MHC expression: 1 degree myotubes coexpress embryonic and slow MHC, while 2 degrees myotubes initially express only embryonic MHC. We have used monoclonal antibodies which recognize the embryonic, slow, neonatal, and adult fast IIB/IIX MHCs to examine MHC accumulation in myoblasts obtained from hindlimbs of embryonic day (ED) 14 and ED 20 Sprague-Dawley rats during differentiation in vitro. Embryonic myoblasts (ED 14), which develop into 1 degree myotubes in vivo, differentiate as myocytes or small myotubes (i.e., 1-4 nuclei) which express both embryonic and slow MHC. They do not accumulate detectable levels of neonatal or adult fast IIB/IIX MHC. Fetal myoblasts, which develop into secondary myotubes in vivo, fuse to form large myotubes (i.e., 10-50 nuclei) and express predominantly embryonic MHC at 3 days in culture. These myotubes accumulate neonatal and adult fast IIB/IIX isoforms of MHC and eventually contract spontaneously. In contrast to embryonic myotubes, they do not accumulate slow MHC. Our results demonstrate that embryonic and fetal rat myoblasts express different phenotypes in vitro and suggest that they represent distinct myoblast lineages similar to those previously described in chickens and mice. These two lineages may be responsible for the generation of distinct populations of 1 degree and 2 degrees myotubes in vivo.