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大鼠成年离体心肌细胞中结蛋白的免疫荧光和免疫金电子显微镜观察

Immunofluorescence and immunogold electron microscopy of desmin in isolated adult heart myocytes of the rat.

作者信息

Saetersdal T, Dalen H, Røli J

机构信息

Institute of Anatomy, University of Bergen, Norway.

出版信息

Histochemistry. 1989;92(6):467-73. doi: 10.1007/BF00524758.

Abstract

Isolated myocytes of the adult mammalian heart are useful for studying cytoskeletal changes during development of irreversible myocardial injuries. Using monoclonal antibodies we have studied the structural organization of desmin in freshly isolated cardiomyocytes from rat hearts. This preparation consists of approximately 85% calcium tolerant rod shaped cells and 15% contracted "square cells" and "round cells" that were initially injured during separation. Cells were quick-frozen at -196 degrees C without any chemical stabilization, cryosectioned and then further processed for immunofluorescence or immunoelectron microscopy. Freshly isolated rod shaped cells exhibit the specific pattern of interfibrillar desmin organization of striated muscle. Furthermore, high resolution immunogold preparations show that desmin in the rod cells occurs in apposition to the edges of the Z-bands as well as closely associated with the plasmalemma. We could find no evidence for the presence of desmin within the Z-band plaques. This organization of desmin is completely absent in the contracted round cells. Thus, already at advanced stages of square cell development, desmin is almost entirely confined to the outer areas of the central filamentous core. We conclude that during the process of square cell contracture, the filamentous desmin contacts with Z-bands and sarcolemma are broken, leading to the unorganized array of desmin in round cells.

摘要

成年哺乳动物心脏的分离心肌细胞对于研究不可逆心肌损伤发展过程中的细胞骨架变化很有用。我们使用单克隆抗体研究了从大鼠心脏新鲜分离的心肌细胞中结蛋白的结构组织。这种制备物由大约85%的耐钙杆状细胞和15%的收缩型“方形细胞”及“圆形细胞”组成,这些“方形细胞”和“圆形细胞”在分离过程中最初受到了损伤。细胞在-196℃下快速冷冻,无需任何化学固定,进行冷冻切片,然后进一步处理用于免疫荧光或免疫电子显微镜观察。新鲜分离的杆状细胞呈现出横纹肌肌原纤维间结蛋白组织的特定模式。此外,高分辨率免疫金制备显示杆状细胞中的结蛋白与Z带边缘相邻,并且与质膜紧密相关。我们没有发现Z带斑内存在结蛋白的证据。在收缩的圆形细胞中完全不存在这种结蛋白组织。因此,在方形细胞发育的晚期阶段,结蛋白几乎完全局限于中央丝状核心的外部区域。我们得出结论,在方形细胞挛缩过程中,丝状结蛋白与Z带和肌膜的接触被破坏,导致圆形细胞中结蛋白排列无序。

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