Thick filament assembly occurs after the formation of a cytoskeletal scaffold.
The development of myofibrils involves the formation of contractile filaments and their assembly into the strikingly regular structure of the sarcomere. We analysed this assembly process in cultured human skeletal muscle cells and in rat neonatal cardiomyocytes by immunofluorescence microscopy using antibodies directed against cytoskeletal and contractile proteins. In particular, the question in which temporal order the respective proteins are integrated into developing sarcomeres was addressed. Although sarcomeric myosin heavy chain is expressed as one of the first myofibrillar proteins, its characteristic A band arrangement is reached at a very late stage. In contrast, titin, then myomesin and finally C-protein (MyBP-C) gradually form a regularly arranged scaffold on stress fiber-like structures (SFLS), on non-striated myofibrils (NSMF) and on nascent striated myofibrils (naSMF). Immediately subsequent to the completion of sarcomere cytoskeleton formation, the labeling pattern of myosin changes from the continuous staining of SFLS to the periodic staining characteristic for mature myofibrils. This series of events can be seen most clearly in the skeletal muscle cell cultures and--probably due to a faster developmental progression less well in cardiomyocytes. We therefore conclude that the correct assembly of a cytoskeletal scaffold is a prerequisite for correct thick filament assembly and for the integration of the contractile apparatus into the myofibril.
Van der Ven
P F
PF
Department of Cell Biology, Institute of Zoophysiology and Cell Biology, University of Potsdam, Germany. pvdven@rz.uni-potsdam.de
Ehler
E
E
Perriard
J C
JC
Fürst
D O
DO
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism; D002478:Cells, Cultured; D064211:Connectin; D003599:Cytoskeleton / Q000378:metabolism*; D000939:Epitopes / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006801:Humans; D018613:Microscopy, Confocal; D009124:Muscle Proteins / Q000032:analysis / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D009206:Myocardium / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D009218:Myosins / Q000032:analysis / Q000378:metabolism*; D011494:Protein Kinases / Q000032:analysis / Q000378:metabolism; D051381:Rats; D012518:Sarcomeres / Q000737:chemistry / Q000378:metabolism
D002352:Carrier Proteins; D064211:Connectin; D000939:Epitopes; D009124:Muscle Proteins; C577285:TTN protein, human; C020723:citrate-binding transport protein; D011494:Protein Kinases; D009218:Myosins