title: Costameres are sites of force transmission to the substratum in adult rat cardiomyocytes.
abstract: Costameres, the vinculin-rich, sub-membranous transverse ribs found in many skeletal and cardiac muscle cells (Pardo, J. V., J. D. Siciliano, and S. W. Craig. 1983. Proc. Natl. Acad. Sci. USA. 80:363-367.) are thought to anchor the Z-lines of the myofibrils to the sarcolemma. In addition, it has been postulated that costameres provide mechanical linkage between the cells' internal contractile machinery and the extracellular matrix, but direct evidence for this supposition has been lacking. By combining the flexible silicone rubber substratum technique (Harris, A. K., P. Wild, and D. Stopak. 1980. Science (Wash. DC). 208:177-179.) with the microinjection of fluorescently labeled vinculin and alpha-actinin, we have been able to correlate the distribution of costameres in adult rat cardiac myocytes with the pattern of forces these cells exert on the flexible substratum. In addition, we used interference reflection microscopy to identify areas of the cells which are in close contact to the underlying substratum. Our results indicate that, in older cell cultures, costameres can transmit forces to the extracellular environment. We base this conclusion on the following observations: (a) adult rat heart cells, cultured on the silicone rubber substratum for 8 or more days, produce pleat-like wrinkles during contraction, which diminish or disappear during relaxation; (b) the pleat-like wrinkles form between adjacent alpha-actinin-positive Z-lines; (c) the presence of pleat-like wrinkles is always associated with a periodic, "costameric" distribution of vinculin in the areas where the pleats form; and (d) a banded or periodic pattern of dark gray or close contacts (as determined by interference reflection microscopy) has been observed in many cells which have been in culture for eight or more days, and these close contacts contain vinculin. A surprising finding is that vinculin can be found in a costameric pattern in cells which are contracting, but not producing pleat-like wrinkles in the substratum. This suggests that additional proteins or posttranslational modifications of known costamere proteins are necessary to form a continuous linkage between the myofibrils and the extracellular matrix. These results confirm the hypothesis that costameres mechanically link the myofibrils to the extracellular matrix. We put forth the hypothesis that costameres are composite structures, made up of many protein components; some of these components function primarily to anchor myofibrils to the sarcolemma, while others form transmembrane linkages to the extracellular matrix.
authors.affiliation: Department of Cell and Developmental Biology, Pennsylvania Muscle Institute, School of Medicine, University of Pennsylvania, Philadelphia 19104-6058.
authors.forename: B A
authors.initials: BA
authors.lastname: Danowski
authors.forename: K
authors.initials: K
authors.lastname: Imanaka-Yoshida
authors.forename: J M
authors.initials: JM
authors.lastname: Sanger
authors.forename: J W
authors.initials: JW
authors.lastname: Sanger
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000502:physiology; D006321:Heart / Q000502:physiology*; D008297:Male; D008845:Microinjections; D008856:Microscopy, Fluorescence; D009206:Myocardium / Q000737:chemistry / Q000166:cytology*; D009210:Myofibrils / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D016596:Vinculin / Q000378:metabolism*
chemical_list: D000199:Actins; D016596:Vinculin