Hoshino S, Ohkoshi N, Ishii A, Kameya S, Takeda S, Shoji S
Department of Neurology, Institute of Clinical Medicine, University of Tsukuba, Japan.
J Muscle Res Cell Motil. 2001;22(2):185-91. doi: 10.1023/a:1010553104341.
The expression of dystrophin and alpha1-syntrophin in rat tibialis anterior muscles were evaluated during a cycle of regeneration after myonecrosis induced by the injection of cardiotoxin. Immunohistochemical studies were performed in cryosections of muscles on days 1, 3, 5, 7, 10, 14, 21 and 28 after injection of cardiotoxin. Western blot analysis was also examined in muscle on days 1, 3, 5, 7, 10, 14, 21 and 28. In immunohistochemical studies, dystrophin was stained weakly at the sarcolemma of some regenerating muscle fibers on day 3, and by day 10 it was stained strongly on almost all regenerating muscle fibers. Alpha1-syntrophin was stained weakly at the sarcolemma of some regenerating fibers on day 5, and by day 14 it was detected on all regenerating muscle fibers. In Western blot analysis, dystrophin (DYS1) and alpha1-syntrophin (alpha1S) were completely absent on day 1. Re-expression of DYS1 and alpha1S was visible by day 5 and accelerated thereafter. The Western blots of DYS1 and alpha1S were densitometrically analyzed on each day. The protein levels on each day were converted to the percentage of the protein level on day 28, which was taken as 100%. From the sequential line based on these data, the following results were obtained on the chronological course of DYS1 and alpha1S. DYS1: 25% of the protein level on day 28 was reached by 3.5 days, 50% was reached by 5.3 days, and 90% was reached by 6.9 days. Alpha1S: 25% of the protein level on day 28 was reached by 4.6 days, 50% was reached by 6.0 days, and 90% was reached by 12.5 days. In this study, DYS1 regenerated earlier than alpha1S at the sarcolemma of regenerating muscle fibers.
在注射心肌毒素诱导大鼠胫前肌发生肌坏死之后的一个再生周期内，对肌营养不良蛋白和α1 - 肌营养不良素在大鼠胫前肌中的表达进行了评估。在注射心肌毒素后的第1、3、5、7、10、14、21和28天，对肌肉的冰冻切片进行免疫组织化学研究。在第1、3、5、7、10、14、21和28天也对肌肉进行了蛋白质免疫印迹分析。在免疫组织化学研究中，第3天时，肌营养不良蛋白在一些再生肌纤维的肌膜上染色较弱，到第10天时，几乎所有再生肌纤维上的染色都很强。α1 - 肌营养不良素在第5天时在一些再生纤维的肌膜上染色较弱，到第14天时在所有再生肌纤维上都能检测到。在蛋白质免疫印迹分析中，第1天时肌营养不良蛋白（DYS1）和α1 - 肌营养不良素（α1S）完全缺失。DYS1和α1S在第5天时可见重新表达，此后加速表达。每天对DYS1和α1S的蛋白质免疫印迹进行光密度分析。将每天的蛋白质水平换算为第28天蛋白质水平的百分比，第28天的蛋白质水平被视为100%。根据这些数据绘制的连续曲线，得出了DYS1和α1S在时间进程上的以下结果。DYS1：在3.5天时达到第28天蛋白质水平的25%，在5.3天时达到50%，在6.9天时达到90%。α1S：在4.6天时达到第28天蛋白质水平的25%，在6.0天时达到50%，在12.5天时达到90%。在本研究中，在再生肌纤维的肌膜上，DYS1比α1S更早再生。
