van der Ven P F, Fürst D O
Department of Cell Biology, University of Potsdam, Germany.
J Muscle Res Cell Motil. 1998 Oct;19(7):767-75. doi: 10.1023/a:1005443203238.
The expression and organization patterns of several myofibrillar proteins were analysed in the putative myofibroblast cell line BHK-21/C13. Although this cell line originates from renal tissue, the majority of the cells express titin. In these cells, titin is, under standard culture conditions, detected in myofibril-like structures (MLSs), where it alternates with non-muscle myosin (NMM). Expression of sarcomeric myosin heavy chain (sMyHC) is observed in a small minority of cells, while other sarcomeric proteins, such as nebulin, myosin binding protein C (MyBP-C), myomesin and M-protein are not expressed at all. By changing the culture conditions in a way equal to conditions that induce differentiation of skeletal muscle cells, a process reminiscent of sarcomerogenesis in vitro is induced. Within one day after the switch to a low-nutrition medium, myofibrillar proteins can be detected in a subset of cells, and after two to five days, all myofibrillar proteins examined are organized in typical sarcomeric patterns. Frequently, cross-striations are visible with phase contrast optics. Transfection of these cells with truncated myomesin fragments showed that a specific part of the myomesin molecule, known to contain a titin-binding site, binds to MLSs, whereas other parts do not. These results demonstrate that this cell line could serve as a powerful model to study the assembly of myofibrils. At the same time, its transfectability offers an invaluable tool for in vivo studies concerning binding properties of sarcomeric proteins.
在假定的肌成纤维细胞系BHK-21/C13中分析了几种肌原纤维蛋白的表达和组织模式。尽管该细胞系起源于肾组织,但大多数细胞表达肌联蛋白。在这些细胞中,在标准培养条件下,肌联蛋白在肌原纤维样结构(MLS)中被检测到,在那里它与非肌肉肌球蛋白(NMM)交替出现。在一小部分细胞中观察到肌节肌球蛋白重链(sMyHC)的表达,而其他肌节蛋白,如伴肌动蛋白、肌球蛋白结合蛋白C(MyBP-C)、肌间蛋白和M蛋白根本不表达。通过以等同于诱导骨骼肌细胞分化的条件改变培养条件,诱导了一个体外类似于肌节形成的过程。在转换到低营养培养基后的一天内,可在一部分细胞中检测到肌原纤维蛋白,在两到五天后,所有检测的肌原纤维蛋白都以典型的肌节模式组织起来。通常,相差光学显微镜下可见横纹。用截短的肌间蛋白片段转染这些细胞表明,肌间蛋白分子中已知含有肌联蛋白结合位点的特定部分与MLS结合,而其他部分则不结合。这些结果表明,该细胞系可作为研究肌原纤维组装的有力模型。同时,其可转染性为关于肌节蛋白结合特性的体内研究提供了一个宝贵的工具。