Chen F, Mottino G, Shin V Y, Frank J S
University of California Los Angeles, School of Medicine, Los Angeles, California 90095-1760, USA.
J Mol Cell Cardiol. 1997 Oct;29(10):2621-9. doi: 10.1006/jmcc.1997.0475.
Ankyrins are a multigene family of proteins that function as adapters between the cytoskeleton and trans-membrane proteins, such as ion channels. Previous studies have shown the linkage between ankyrin and ionic transport proteins such as Na+-K+ ATPase, voltage-dependent Na+ channels and Ca2+ channels. In the present study, we have investigated the subcellular distribution of ankyrin and its relationship to the Na+-Ca2+ exchange protein in immature and adult rabbit ventricular myocytes. Isolated single cardiomyocytes from neonatal, juvenile and adult rabbit hearts were examined by immunofluorescence labeling techniques, using antibodies against ankyrin and the Na+-Ca2+ exchanger. We found that in neonatal rabbit cardiac myocytes, ankyrin labeling was mainly present at the Z disk, whereas the Na+-Ca2+ exchanger was only present on the peripheral sarcolemma. At 2 weeks of age, ankyrin labeling was still predominantly observed at the level of the Z disks as well as in the partially developed T-tubules. In the adult cells, however, ankyrin and the Na+-Ca2+ exchanger seem to be co-localized within T-tubules and at the costamere region of the peripheral sarcolemma. Immunogold labeling studies at the higher resolution electron microscopic level using cyrosection tissues of rabbit heart at different ages confirm these findings. These results indicate that the distribution pattern of ankyrin and the Na+-Ca2+ exchanger changes with development in rabbit ventricular myocytes.
锚蛋白是一个多基因家族的蛋白质,其功能是作为细胞骨架与跨膜蛋白(如离子通道)之间的衔接子。先前的研究已经表明锚蛋白与离子转运蛋白(如钠钾ATP酶、电压依赖性钠通道和钙通道)之间存在联系。在本研究中,我们调查了未成熟和成年兔心室肌细胞中锚蛋白的亚细胞分布及其与钠钙交换蛋白的关系。使用针对锚蛋白和钠钙交换体的抗体,通过免疫荧光标记技术对从新生、幼年和成年兔心脏分离出的单个心肌细胞进行了检测。我们发现,在新生兔心肌细胞中,锚蛋白标记主要存在于Z线,而钠钙交换体仅存在于外周肌膜上。在2周龄时,仍主要在Z线水平以及部分发育的T小管中观察到锚蛋白标记。然而,在成年细胞中,锚蛋白和钠钙交换体似乎共定位于T小管内以及外周肌膜的肌附着点区域。使用不同年龄兔心脏的冷冻切片组织在更高分辨率的电子显微镜水平上进行的免疫金标记研究证实了这些发现。这些结果表明,兔心室肌细胞中锚蛋白和钠钙交换体的分布模式随发育而变化。