Sweeney L J, Kennedy J M, Zak R, Kokjohn K, Kelley S W
Department of Anatomy, Loyola University Medical Center, Maywood, Illinois 60153.
Dev Biol. 1989 Jun;133(2):361-74. doi: 10.1016/0012-1606(89)90040-7.
We have utilized a key biochemical determinant of muscle fiber type, myosin isoform expression, to investigate the initial developmental program of future fast and slow skeletal muscle fibers. We examined myosin heavy chain (HC) phenotype from the onset of myogenesis in the limb bud muscle masses of the chick embryo through the differentiation of individual fast and slow muscle masses, as well as in newly formed myotubes generated in adult muscle by weight overload. Myosin HC isoform expression was analyzed by immunofluorescence localization with a battery of anti-myosin antibodies and by electrophoretic separation with SDS-PAGE. Results showed that the initial myosin phenotype in all skeletal muscle cells formed during the embryonic period (until at least 8 days in ovo) consisted of expression of a myosin HC which shares antigenic and electrophoretic migratory properties with ventricular myosin and a distinct myosin HC which shares antigenic and electrophoretic migratory properties with fast skeletal isomyosin. Similar results were observed in newly formed myotubes in adult muscle. Future fast and slow muscle fibers could only be discriminated from each other in developing limb bud muscles by the onset of expression of slow skeletal myosin HC at 6 days in ovo. Slow skeletal myosin HC was expressed only in myotubes which became slow fibers. These findings suggest that the initial commitment of skeletal muscle progenitor cells is to a common skeletal muscle lineage and that commitment to a fiber-specific lineage may not occur until after localization of myogenic cells in appropriate premuscle masses. Thus, the process of localization, or events which occur soon thereafter, may be involved in determining fiber type.
我们利用肌肉纤维类型的一个关键生化决定因素——肌球蛋白同工型表达,来研究未来快、慢骨骼肌纤维的初始发育程序。我们从鸡胚肢体芽肌肉块的肌发生开始,通过个体快、慢肌肉块的分化,以及成年肌肉因重量过载产生的新形成的肌管,研究了肌球蛋白重链(HC)表型。通过使用一系列抗肌球蛋白抗体进行免疫荧光定位以及通过SDS-PAGE进行电泳分离来分析肌球蛋白HC同工型的表达。结果表明,在胚胎期形成的所有骨骼肌细胞(至少直到胚胎发育8天)中的初始肌球蛋白表型包括一种与心室肌球蛋白具有共同抗原性和电泳迁移特性的肌球蛋白HC的表达,以及一种与快骨骼肌异肌球蛋白具有共同抗原性和电泳迁移特性的独特肌球蛋白HC的表达。在成年肌肉新形成的肌管中也观察到了类似的结果。在发育中的肢体芽肌肉中,直到胚胎发育6天时慢骨骼肌肌球蛋白HC表达开始,未来的快、慢肌纤维才能相互区分。慢骨骼肌肌球蛋白HC仅在那些成为慢纤维的肌管中表达。这些发现表明,骨骼肌祖细胞的初始定向是朝着共同的骨骼肌谱系,并且对纤维特异性谱系的定向可能直到肌源性细胞在适当的肌肉前体块中定位之后才会发生。因此,定位过程或此后不久发生的事件可能参与决定纤维类型。