Contractile activity and cell-cell contact regulate myofibrillar organization in cultured cardiac myocytes.
Adult feline ventricular myocytes cultured on a laminin-coated substratum reestablish intercellular junctions, yet disassemble their myofibrils. Immunofluorescence microscopy reveals that these non-beating heart cells lack vinculin-positive focal adhesions; moreover, intercellular junctions are also devoid of vinculin. When these quiescent myocytes are stimulated to contract with the beta-adrenergic agonist, isoproterenol, extensive vinculin-positive focal adhesions and intercellular junctions emerge. If solitary myocytes are stimulated to beat, an elaborate series of vinculin-positive focal adhesions develop which appear to parallel the reassembly of myofibrils. In cultures where neighboring myocytes reestablish cell-cell contact, myofibrils appear to reassemble from the fascia adherens rather than focal contacts. Activation of beating is accompanied by a significant reduction in the rate of total and cytoskeletal protein synthesis; in fact, myofibrillar reassembly, redevelopment of focal adhesions and fascia adherens junctions require no protein synthesis for at least 24 h, implying the existence of an assembly competent pool of cytoskeletal proteins. Maturation of the fasciae adherens and the appearance of vinculin within Z-line/costameres, does require de novo synthesis of new cytoskeletal proteins. Changes in cytoskeletal protein turnover appear dependent on beta agonist-induced cAMP production, but myofibrillar reassembly is a cAMP-independent event. Such observations suggest that mechanical forces, in the guise of contractile activity, regulate vinculin distribution and myofibrillar order in cultured adult feline heart cells.
Simpson
D G
DG
Department of Cell, Molecular and Structural Biology, Northwestern University Medical School, Chicago, Illinois 60611.
Decker
M L
ML
Clark
W A
WA
Decker
R S
RS
D000199:Actins / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002415:Cats; D002450:Cell Communication / Q000502:physiology*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism / Q000502:physiology; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D007365:Intercellular Junctions / Q000502:physiology / Q000648:ultrastructure; D007545:Isoproterenol / Q000494:pharmacology; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009200:Myocardial Contraction / Q000502:physiology*; D009206:Myocardium / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D009210:Myofibrils / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D016596:Vinculin / Q000032:analysis / Q000378:metabolism
D000199:Actins; D003598:Cytoskeletal Proteins; D016596:Vinculin; D000242:Cyclic AMP; D007545:Isoproterenol