Lee-De Groot M B, Van der Laarse W J
Laboratory for Physiology, Institute for Cardiovascular Research (ICaR-VU), Amsterdam, The Netherlands.
J Muscle Res Cell Motil. 1996 Aug;17(4):439-48. doi: 10.1007/BF00123360.
Mature, high-oxidative, skeletal muscle fibres of Xenopus laevis were kept in culture in L-(Leibovitz's)-15 medium supplemented with creatine and antibiotics and some other additions. Single fibres were mounted at a fixed sarcomere length in a flow-through culture chamber which accommodates stimulus electrodes and a force transducer. Twitch characteristics were determined daily. Depending on culture conditions, fibres remained excitable electrically for up to two weeks at 20 degrees C when foetal calf serum and/or phosphate were added to the culture medium. During the second week, fibres lost phosphocreatine and ATP, but relatively small changes (if any) in total creatine, glycogen and protein contents, fibre volume and dry weight occurred. Succinate dehydrogenase activity decreased after 9 days-when ATP was reduced already. Fibres which were inexcitable electrically contracted normally when exposed to caffeine, indicating that excitation-contraction coupling failed and that the contractile apparatus was still functional.
将非洲爪蟾成熟的、高氧化型骨骼肌纤维置于添加了肌酸、抗生素及其他一些添加物的L-(莱博维茨)-15培养基中进行培养。将单根纤维以固定的肌节长度安装在一个流通式培养室中,该培养室可容纳刺激电极和力传感器。每天测定肌肉收缩特性。根据培养条件,当向培养基中添加胎牛血清和/或磷酸盐时,纤维在20℃下可保持长达两周的电兴奋性。在第二周,纤维失去磷酸肌酸和三磷酸腺苷(ATP),但总肌酸、糖原和蛋白质含量、纤维体积和干重发生相对较小的变化(如有变化)。琥珀酸脱氢酶活性在第9天(此时ATP已经减少)后下降。电兴奋性丧失的纤维在暴露于咖啡因时仍能正常收缩,这表明兴奋-收缩偶联失败,但收缩装置仍有功能。