Distribution of fast myosin heavy chain isoforms in thick filaments of developing chicken pectoral muscle.
Colloidal gold-conjugated monoclonal antibodies were prepared to stage-specific fast myosin heavy chain (MHC) isoforms of developing chicken pectoralis major (PM). Native thick filaments from different stages of development were reacted with these antibodies and examined in the electron microscope to determine their myosin isoform composition. Filaments prepared from 12-d embryo, 10-d chick, and 1-yr chicken muscle specifically reacted with the embryonic (EB165), neonatal (2E9), and adult (AB8) antimyosin gold-conjugated monoclonal antibodies, respectively. The myosin isoform composition was more complex in thick filaments from stages of pectoral muscle where more than one isoform was simultaneously expressed. In 19-d embryo muscle where both embryonic and neonatal isoforms were present, three classes of filaments were found. One class of filaments reacted only with the embryonic antibody, a second class reacted only with the neonatal-specific antibody, and a third class of filaments were decorated by both antibodies. Similar results were obtained with filaments prepared from 44-d chicken PM where the neonatal and adult fast MHCs were expressed. These observations demonstrate that two myosin isoforms can exist in an individual thick filament in vivo. Immunoelectron microscopy was also used to determine the specific distribution of different fast MHC isoforms within individual filaments from different stages of development. The anti-embryonic and anti-adult antibodies uniformly decorated both homogeneous and heterogeneous thick filaments. The neonatal specific antibody uniformly decorated homogeneous filaments; however, it preferentially decorated the center of heterogeneous filaments. These observations suggest that neonatal MHC may play a specific role in fibrillogenesis.
Taylor
L D
LD
Department of Food Science and Technology, University of California, Davis 95616.
Bandman
E
E
D000818:Animals; D000911:Antibodies, Monoclonal; D002642:Chick Embryo; D002645:Chickens; D006046:Gold; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D024510:Muscle Development; D009132:Muscles / Q000032:analysis / Q000196:embryology; D009218:Myosins / Q000032:analysis*; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D006046:Gold; D009218:Myosins