Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro.
Monoclonal antibodies (McAbs) against the myosin heavy chain (MHC) of adult chicken pectoralis muscle have been tested for reactivity with pectoralis myosin at selected stages of chick development in vivo and in vitro. Three such McAbs, MF 20 and MF 14, which bind to light meromyosin, and MF 30, which binds to myosin subfragment two (S2), were used to assay the appearance and accumulation of specific MHC epitopes with: (a) indirect, solid phase radioimmune assay (RIA), (b) immunoautoradiography, (c) immunofluorescence microscopy. McAb MF 20 bound strongly and equivalently to MHC at all stages of embryonic development in vivo. In contrast, the MF 30 epitope was barely detectable at 12 d of incubation but its concentration rose rapidly just before hatching. No detectable binding of MF 14 to pectoralis myosin could be measured during myogenesis in vivo until 1 wk after hatching. Immunofluorescence studies revealed that all three epitopes accumulate in the same myocytes of the developing pectoralis muscle. Since all three McAbs bound with high activity to native and denatured forms of myosin, it is unlikely that differential antibody reactivity can be explained by conformational changes in myosin during development in vivo. When myogenesis in vitro was monitored using the same McAbs, MF 20 bound to the MHC at all stages tested while reactivity of MF 30 and MF 14 with myosin from cultured muscle was never observed. Thus, this study demonstrates three different immunochemical states of the MHC during development in vivo of chick pectoralis muscle and the absence of later occurring immunochemical transitions in the MHC of cultured embryonic muscle.
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D000939:Epitopes / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D024510:Muscle Development; D009132:Muscles / Q000032:analysis; D015879:Myosin Subfragments / Q000276:immunology; D009218:Myosins / Q000032:analysis / Q000276:immunology*; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015879:Myosin Subfragments; D009218:Myosins