运用快速冷冻和深度蚀刻法对培养的小鼠肌肉细胞中的抗肌萎缩蛋白进行免疫细胞化学研究。

Immunocytochemical study of dystrophin in cultured mouse muscle cells by the quick-freezing and deep-etching method.

作者信息

Park-Matsumoto Y C, Ohno S, Baba T, Kobayashi T, Tsukagoshi H

机构信息

Department of Neurology, Tokyo Medical and Dental University School of Medicine, Japan.

出版信息

Histochem J. 1992 Jun;24(6):383-92. doi: 10.1007/BF01046171.

DOI:10.1007/BF01046171

PMID:1634376

Abstract

Dystrophin, the protein product of the Duchenne muscular dystrophy (DMD) gene, is deficient in patients with DMD and in mdx mice. It is immunocytochemically localized in skeletal muscle sarcolemma. However, little is known about the three-dimensional ultrastructural localization of dystrophin and its relationship with other cytoskeletal proteins. We found that dystrophin is localized irregularly, just underneath the plasma membrane in normal cultured mouse myotubes, by using the quick-freezing and deep-etching (QF-DE) method; it was found to be closely linked to actin-like filaments (8-10 nm in diameter), most of which were decorated with myosin subfragment 1, and was attached to the cytoplasmic side of the plasma membrane. These results suggest that dystrophin might play an important role in the preservation of cell membrane stability by connecting actin cytoskeletons with the cytoplasmic side of the plasma membrane.

摘要

肌营养不良蛋白是杜兴氏肌营养不良症（DMD）基因的蛋白质产物，在DMD患者和mdx小鼠中缺乏。它通过免疫细胞化学定位在骨骼肌肌膜中。然而，关于肌营养不良蛋白的三维超微结构定位及其与其他细胞骨架蛋白的关系知之甚少。我们发现，通过快速冷冻和深度蚀刻（QF-DE）方法，在正常培养的小鼠肌管中，肌营养不良蛋白不规则地定位在质膜下方；发现它与肌动蛋白样细丝（直径8-10纳米）紧密相连，其中大部分被肌球蛋白亚片段1修饰，并附着在质膜的细胞质侧。这些结果表明，肌营养不良蛋白可能通过将肌动蛋白细胞骨架与质膜的细胞质侧连接起来，在维持细胞膜稳定性方面发挥重要作用。