Myofibril assembly is linked with vinculin, alpha-actinin, and cell-substrate contacts in embryonic cardiac myocytes in vitro.
The relationship of nascent myofibrils with the accumulation of adhesion plaque proteins and the formation of focal cell contacts was studied in embryonic chick cardiac myocytes in vitro. The cultures were double-stained with various combinations of the specific antiactin drug phalloidin and antibodies against vinculin, alpha-actinin, connectin (titin), myosin heavy chain, fibronectin, and desmin and examined under fluorescence and interference reflection microscopy. In the areas of myofibril assembly, vinculin and alpha-actinin plaques were formed at the ventral sarcolemmae. These areas overlapped with the sites of cell-to-substrate focal contacts and extracellular fibronectin. Because the myofibrils always ran in a straight line between these sites, polarized lines appeared to be generated within the cells in response to their physical (e.g., stress) and/or biochemical environment (e.g., adhesion plaque proteins). The possible presence of other factors cannot be ruled out for the proper alignment of myofibrils. As soon as myofibrils came to span between these adhesion sites, they exhibited typically mature cross-striated characteristics. Thus, the formation of these inferred lines has some relation to, or is in fact necessary for, the maturation of myofibrils, in addition to the directional arrangement of sarcomeric proteins. Additionally, synthesis and distribution of myosin and connectin were tightly linked during early developmental (premyofibril and myofibril) stages. The spatial deployment of desmin was not coupled with vinculin. Thus, connectin and desmin do not appear to form the initial scaffold of sarcomeres.
Terai
M
M
Department of Anatomy, School of Medicine, Chiba University, Japan.
Komiyama
M
M
Shimada
Y
Y
D000185:Actinin / Q000032:analysis*; D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D064211:Connectin; D003893:Desmin / Q000032:analysis; D005353:Fibronectins / Q000032:analysis; D006321:Heart / Q000196:embryology; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000032:analysis; D008856:Microscopy, Fluorescence; D008857:Microscopy, Interference; D009124:Muscle Proteins / Q000032:analysis*; D009210:Myofibrils / Q000032:analysis*; D009218:Myosins / Q000032:analysis; D011494:Protein Kinases; D016596:Vinculin
D064211:Connectin; D003893:Desmin; D005353:Fibronectins; D008565:Membrane Proteins; D009124:Muscle Proteins; D000185:Actinin; D016596:Vinculin; D011494:Protein Kinases; D009218:Myosins