Zs-Nagy I, Zhang X, Kitani K, Nonomura Y
Verzár International Laboratory for Experimental Gerontology (VILEG), University Medical School, Debrecen, Hungary.
Biochem Biophys Res Commun. 1995 Oct 4;215(1):67-74. doi: 10.1006/bbrc.1995.2434.
The expression of dystrophin can be suppressed in cultured skeletal muscle cells (coded L-185 from rat, and C2 from mouse) after a proper genetic manipulation. The influence of presence or absence of dystrophin on the lateral diffusion constant of Con-A-receptors was studied in the cell membrane of such cells (and also of mature skeletal muscle fibres of rat and mouse) by means of the fluorescence recovery after photobleaching (FRAP) technique, applying a novel fluorescent label called Con-A-BODIPY-FI conjugate. It has been established that the normal maturation, of myoblasts into skeletal muscle fibres involves a significant decrease of the mobility of Con-A receptors in the sarcolemma. In the absence of dystrophin, this maturation process cannot take place; the membrane proteins display an increasing mobility during the culture time, which is of lethal effect for these cells.
经过适当的基因操作后，培养的骨骼肌细胞（大鼠的编码为L-185，小鼠的为C2）中肌营养不良蛋白的表达可被抑制。利用光漂白后荧光恢复（FRAP）技术，应用一种名为Con-A-硼二吡咯氟化物（Con-A-BODIPY-FI）偶联物的新型荧光标记，研究了此类细胞（以及大鼠和小鼠的成熟骨骼肌纤维）细胞膜中有无肌营养不良蛋白对刀豆球蛋白A（Con-A）受体侧向扩散常数的影响。已经确定，成肌细胞正常成熟为骨骼肌纤维涉及肌膜中Con-A受体流动性的显著降低。在缺乏肌营养不良蛋白的情况下，这种成熟过程无法发生；膜蛋白在培养期间表现出越来越高的流动性，这对这些细胞具有致命影响。
