Capote Joana, Bolaños Pura, Schuhmeier Ralph Peter, Melzer Werner, Caputo Carlo
[corrected] Instituto Venezolano de Investigaciones Cientificas IVIC, Apartado 21827, Caracas 1020A, Venezuela.
J Physiol. 2005 Apr 15;564(Pt 2):451-64. doi: 10.1113/jphysiol.2004.081034. Epub 2005 Feb 24.
Ca(2)(+) transients elicited by action potentials were measured using MagFluo-4, at 20-22 degrees C, in intact muscle fibres enzymatically dissociated from mice of different ages (7, 10, 15 and 42 days). The rise time of the transient (time from 10 to 90% of the peak) was 2.4 and 1.1 ms in fibres of 7- and 42-day-old mice, respectively. The decay of the transient was described by a double exponential function, with time constants of 1.8 and 16.4 ms in adult, and of 4.6 and 105 ms in 7-day-old animals. The fractional recovery of the transient peak amplitude after 10 ms, F(2(10))/F(1), determined using twin pulses, was 0.53 for adult fibres and ranged between 0.03 and 0.60 in fibres of 7-day-old animals This large variance may indicate differences in the extent of inactivation of Ca(2)(+) release, possibly related to the difference in ryanodine receptor composition between young and old fibres. At the 7 and 10 day stages, fibres responded to Ca(2)(+)-free solutions with a larger decrease in the transient peak amplitude (25% versus 11% in adult fibres), possibly indicating a contribution of Ca(2)(+) influx to the Ca(2)(+) transient in younger animals. Cyclopiazonic acid (1 mum), an inhibitor of the sarcoplasmic reticulum (SR) Ca(2)(+)-ATPase, abolished the Ca(2)(+) transient decay in fibres of 7- and 10-day-old animals and significantly reduced its rate in older animals. Analysis of the transients with a Ca(2)(+) removal model showed that the results are consistent with a larger relative contribution of the SR Ca(2)(+) pump and a lower expression of myoplasmic Ca(2)(+) buffers in fibres of young versus old animals.
在20 - 22摄氏度下,使用MagFluo - 4在从不同年龄(7、10、15和42天)小鼠酶解获得的完整肌纤维中测量动作电位引发的Ca(2)(+)瞬变。瞬变的上升时间(从峰值的10%到90%的时间)在7日龄和42日龄小鼠的纤维中分别为2.4毫秒和1.1毫秒。瞬变的衰减由双指数函数描述,成年动物的时间常数为1.8毫秒和16.4毫秒,7日龄动物的时间常数为4.6毫秒和105毫秒。使用双脉冲测定的10毫秒后瞬变峰值幅度的分数恢复率F(2(10))/F(1),成年纤维为0.53,7日龄动物的纤维在0.03至0.60之间。这种较大的差异可能表明Ca(2)(+)释放的失活程度不同,可能与幼龄和老龄纤维中兰尼碱受体组成的差异有关。在7日龄和10日龄阶段,纤维对无Ca(2)(+)溶液的反应是瞬变峰值幅度有更大程度的下降(成年纤维为11%,幼龄动物纤维为25%),这可能表明Ca(2)(+)内流对幼龄动物Ca(2)(+)瞬变有贡献。肌浆网(SR)Ca(2)(+) - ATP酶抑制剂环匹阿尼酸(1μM)消除了7日龄和10日龄动物纤维中的Ca(2)(+)瞬变衰减,并显著降低了老龄动物纤维中的衰减速率。用Ca(2)(+)去除模型对瞬变进行分析表明,结果与幼龄动物纤维中SR Ca(2)(+)泵的相对贡献更大以及肌浆Ca(2)(+)缓冲蛋白的表达较低一致。