Carrino D A, Oron U, Pechak D G, Caplan A I
Department of Biology, Case Western Reserve University, Cleveland, Ohio 44106.
Development. 1988 Aug;103(4):641-56. doi: 10.1242/dev.103.4.641.
Previous work from this laboratory involved the characterization of a large chondroitin sulphate proteoglycan unique to chick skeletal muscle. This proteoglycan is synthesized by embryonic skeletal muscle both in ovo and in culture but is not synthesized by adult muscle in vivo and myotubes in advanced cultures. Because regenerating skeletal muscle has been found to recapitulate synthesis of embryonic muscle protein isoforms, an analysis was performed to assess whether synthesis of chondroitin sulphate proteoglycans is reinitiated during muscle regeneration. Adult chicken pectoral and leg (gastrocnemius) muscle was injured by excision of a small piece of tissue or by cold injury; in the latter, the basement membrane has been reported to remain intact. At various times after injury, whole animals were exposed to [35S]sulphate and the proteoglycans were isolated by ion-exchange chromatography and analysed. Synthesis of only small proteoglycans, typical of normal adult skeletal muscle, is observed in the contralateral, uninjured muscle. In the regenerating muscle 4 days after injury, there is increased sulphate incorporation and abundant synthesis of chondroitin sulphate proteoglycans. This is observed in both pectoral and leg muscle irrespective of the type of injury, which suggests that the presence of basement membrane does not affect reinitiation of chondroitin sulphate proteoglycan synthesis. By 25 days after injury, synthesis of chondroitin sulphate proteoglycans is still detected but is significantly diminished. These data are consistent with the notion that skeletal muscle regeneration involves a recapitulation of embryonic events and give further credence to the hypothesis that skeletal muscle chondroitin sulphate proteoglycans play a role in some early aspect of myogenesis.
本实验室之前的工作涉及对一种鸡骨骼肌特有的大型硫酸软骨素蛋白聚糖的特性研究。这种蛋白聚糖由胚胎骨骼肌在胚胎发育过程中和培养条件下合成,但成年肌肉在体内以及晚期培养的肌管中均不合成。由于已发现再生骨骼肌会重现胚胎肌肉蛋白异构体的合成过程,因此进行了一项分析,以评估硫酸软骨素蛋白聚糖的合成在肌肉再生过程中是否会重新启动。成年鸡的胸肌和腿部(腓肠肌)通过切除一小块组织或冷损伤的方式造成损伤;在冷损伤的情况下,据报道基底膜保持完整。在损伤后的不同时间,将整只动物暴露于[35S]硫酸盐中,然后通过离子交换色谱法分离蛋白聚糖并进行分析。在对侧未受伤的肌肉中,仅观察到正常成年骨骼肌典型的小型蛋白聚糖的合成。在损伤后4天的再生肌肉中,硫酸盐掺入增加,且硫酸软骨素蛋白聚糖大量合成。无论损伤类型如何,在胸肌和腿部肌肉中均观察到这种情况,这表明基底膜的存在并不影响硫酸软骨素蛋白聚糖合成的重新启动。到损伤后25天,仍可检测到硫酸软骨素蛋白聚糖的合成,但显著减少。这些数据与骨骼肌再生涉及胚胎事件重现的观点一致,并进一步支持了骨骼肌硫酸软骨素蛋白聚糖在肌发生的某些早期阶段起作用的假说。