Immunofluorescent studies for alpha-actinin in cultured cardiomyopathic hamster heart cells

Primary cultures of cardiac myocytes from normal and genetically cardiomyopathic (CM) newborn hamsters (strain UM X7.1) were analyzed by indirect immunofluorescent microscopy after 3, 5, 7, and 9 days in culture. The cultures were fixed in cold acetone and immunostained by an indirect method using FITC-labelled anti-alpha-actinin to label the myofibrillar Z bands. Most normal and CM myocytes appeared round in shape after 3 days in culture. Normal cardiac myocytes began to exhibit cytoplasmic projections after 5 days in culture and their myofibrils usually showed parallel arrangements with respect to each other. The cardiac cells from CM hearts showed an obvious myofibril disarray. Moreover, projections formed later than normal. As the size of the cells increased, more and more projections formed in normal hamster myocytes during development. By contrast, most of the cardiomyopathic myocytes showed few projections even as late as 9 days in culture. Hence, the number of projections per cell was much less in cardiomyopathic myocytes than in normal, especially after 7 and 9 days in culture. These results suggest that cardiomyopathic cells have abnormal shapes in culture and, in particular, fail to form projections as in normal cells. Whether this unusual behavior is related to an abnormality of the membranes or cytoskeletal system in cardiomyopathic heart cells or to some other factor requires further study.