Yoles E, Bak A, Sampson S R
Department of Life Sciences, Bar-Ilan University, Ramat-Gan 52100, Israel.
Int J Dev Neurosci. 1984;2(5):483-90. doi: 10.1016/0736-5748(84)90050-9.
Serum-free, chemically defined media have been reported to provide suitable conditions for growth and proliferation of mammalian skeletal muscle, but there is no information regarding the ability of myotubes to develop normal electrophysiological properties in these media. We have recorded transmembrane resting and action potentials from rat skeletal myotubes grown in both serum-containing (GM) and serum-free chemically defined (CDM) growth media. Muscle cells in CDM do not develop as high resting Em as their counterparts in conventional growth media. CDM myotubes also had a lower incidence and frequency of spontaneously occurring action potentials. Treatment with ouabain or decrease in temperature of the recording medium reduced resting Em of both GM and CDM cells to the same level. We found that the sensitivity of CDM cells to ouabain was about 10-fold higher than that of GM cells. An increase in temperature of the recording medium increased Em of GM myotubes but not of CDM myotubes. The change in resting Em in response to a 10-fold change in extracellular K(+)-ion concentration was the same for both groups of cells thus indicating that there was no difference in membrane permeability to K(+)-ion. We conclude that the difference in Em can be accounted for largely, if not entirely, by differences in activity or amount of electrogenic Na(+)-K(+) ATPase.
据报道,无血清、化学成分明确的培养基可为哺乳动物骨骼肌的生长和增殖提供适宜条件,但尚无关于肌管在这些培养基中发育出正常电生理特性能力的相关信息。我们记录了在含血清(GM)和无血清化学成分明确(CDM)生长培养基中生长的大鼠骨骼肌肌管的跨膜静息电位和动作电位。CDM中的肌肉细胞静息膜电位(Em)不如传统生长培养基中的细胞高。CDM肌管自发动作电位的发生率和频率也较低。用哇巴因处理或降低记录培养基的温度可使GM和CDM细胞的静息Em降至相同水平。我们发现,CDM细胞对哇巴因的敏感性比GM细胞高约10倍。记录培养基温度升高会使GM肌管的Em增加,但不会使CDM肌管的Em增加。两组细胞对细胞外K⁺离子浓度10倍变化时静息Em的变化相同,这表明两组细胞对K⁺离子的膜通透性没有差异。我们得出结论,Em的差异如果不是完全由生电Na⁺-K⁺ ATP酶的活性或数量差异导致,那么很大程度上也可以由这些差异来解释。