Localization of annexin V in the adult and neonatal heart

Annexins are a major family of intracellular Ca2+-binding proteins which have been implicated in a variety of cellular functions. Several conflicting reports have been published on the location of annexin V in the heart. In this paper we have used confocal microscopy to demonstrate that annexin V is associated with the sarcolemma and intercalated discs of cardiac myocytes in sections of adult porcine and rat heart. In addition, we have used confocal microscopy of isolated rat myocytes to show that this association is stable even after cells were treated with the intracellular calcium chelator BAPTA-AM, to reduce cytosolic calcium levels to very low levels. This demonstrates that annexin V associates tightly with the sarcolemma and suggests that components in addition to phospholipid are involved in binding annexin V to the membrane. Furthermore, we show that, in sections of the neonatal rat left ventricle, annexin V has a different subcellular location than that observed in the terminally differentiated adult myocyte. In these differentiating neonatal cells, annexin V is also located in the nucleoplasm and at the periphery of the nucleus. These results demonstrate that the subcellular location of annexin V is differentially regulated and suggest that annexin V regulates calcium-dependent processes at both the sarcolemma and the nucleus.