Obara K, Yamada T
Jpn J Physiol. 1984;34(6):1089-104. doi: 10.2170/jjphysiol.34.1089.
Properties of isolated single smooth muscle cells in which the surface membrane has been made highly permeable by saponin treatment are described. The single cells were isolated from guinea pig taenia caeci by digestion with collagenase and skinned in a relaxing solution by treatment with 7 micrograms/ml saponin for 10 min. The skinned single cells showed Ca2+-regulated shortening in the presence of Mg-ATP, and the maximum degree of shortening was larger than that of the potassium-induced shortening observed in the intact isolated single cells. The half maximum shortening of the skinned single cells occurred at about 5 x 10(-8) Ca2+. The skinned single cells showed a significantly higher Ca2+-sensitivity than the skinned fiber bundles. The shortening-pCa curve for the skinned single cells was unchanged by alteration of pH and ionic strength, but it was shifted to the left by increasing MgATP concentration or to the right by increasing free Mg2+ concentration. The skinned single cells retained their internal Ca2+ storage site function. Caffeine induced shortening in the skinned single cells preloaded with Ca2+, and this shortening was suppressed by procaine. The release of Ca2+ from the storage site could be produced and facilitated by Ca2+ itself when the skinned single cells were exposed to Ca2+ with a concentration of about 2 x 10(-8) M and this release was suppressed by procaine. These results suggest that the Ca2+-induced Ca2+ release mechanism may play a role in the regulation of the stored Ca2+ in this cell.
本文描述了经皂角苷处理使表面膜具有高通透性的分离单个平滑肌细胞的特性。通过用胶原酶消化从豚鼠盲肠带分离出单个细胞,并用7微克/毫升皂角苷处理10分钟,使其在松弛溶液中去膜。去膜的单个细胞在存在Mg-ATP的情况下表现出Ca2+调节的收缩,并且最大收缩程度大于在完整分离的单个细胞中观察到的钾诱导的收缩。去膜单个细胞的最大收缩一半时的Ca2+浓度约为5×10(-8)。去膜单个细胞显示出比去膜纤维束显著更高的Ca2+敏感性。去膜单个细胞的收缩-pCa曲线不受pH和离子强度改变的影响,但通过增加MgATP浓度向左移动,或通过增加游离Mg2+浓度向右移动。去膜单个细胞保留了其内部Ca2+储存位点功能。咖啡因诱导预先加载Ca2+的去膜单个细胞收缩,并且这种收缩被普鲁卡因抑制。当去膜单个细胞暴露于浓度约为2×10(-8)M的Ca2+时,Ca2+本身可以产生并促进Ca2+从储存位点的释放,并且这种释放被普鲁卡因抑制。这些结果表明,Ca2+诱导的Ca2+释放机制可能在该细胞中储存Ca2+的调节中起作用。