Imaizumi Y, Henmi S, Uyama Y, Atsuki K, Torii Y, Ohizumi Y, Watanabe M
Department of Chemical Pharmacology, Faculty of Pharmaceutical Sciences, Nagoya City University, Japan.
Am J Physiol. 1996 Sep;271(3 Pt 1):C772-82. doi: 10.1152/ajpcell.1996.271.3.C772.
Characteristics of Ca2+ release from stores were investigated in strips from ileum and portal vein and in isolated myocytes from ileum and urinary bladder of the guinea pig with use of caffeine and 9-methyl-7-bromoeudistomin D (MBED), a potent releaser of Ca2+ from skeletal muscle sarcoplasmic reticulum. In skinned strips, 1-30 mM caffeine elicited a transient contraction, but 10-300 microM MBED did not. Pretreatment with 100 microM MBED did not affect the subsequent caffeine-induced contraction. In single cells loaded with indo 1-acetoxymethyl ester, 10 mM caffeine increased cytoplasmic Ca2+ concentration, whereas 100 microM MBED elicited a small or no increase. Under whole cell clamp, spontaneous transient outward currents through Ca(2+)-dependent K+ (BK) channels were first enhanced and then suppressed by 30 microM MBED or 5 mM caffeine. The amplitude of Ca(2+)-dependent transient K+ current on depolarization was reduced by MBED and caffeine (50% inhibitory concentrations = 20 microM and 1 mM, respectively). Other currents and single BK channel activity were not significantly affected by MBED. The Ca2+ release from stores responsible for BK channel activation may be resolved from that for the activation of the contractile system by MBED in these smooth muscle cells.
利用咖啡因和9-甲基-7-溴海兔毒素D(MBED,一种从骨骼肌肌浆网释放Ca2+的强效物质),对豚鼠回肠和门静脉条带以及回肠和膀胱分离的肌细胞中储存Ca2+释放的特性进行了研究。在剥除肌膜的条带中,1-30 mM咖啡因引起短暂收缩,但10-300 microM MBED未引起收缩。用100 microM MBED预处理不影响随后咖啡因诱导的收缩。在加载indo 1-乙酰氧基甲酯的单细胞中,10 mM咖啡因增加细胞质Ca2+浓度,而100 microM MBED引起少量增加或无增加。在全细胞钳制下,通过Ca(2+)依赖性K+(BK)通道的自发性短暂外向电流首先被30 microM MBED或5 mM咖啡因增强,然后被抑制。去极化时Ca(2+)依赖性短暂K+电流的幅度被MBED和咖啡因降低(50%抑制浓度分别为20 microM和1 mM)。其他电流和单个BK通道活性未受到MBED的显著影响。在这些平滑肌细胞中,负责BK通道激活的储存Ca2+释放可能与MBED激活收缩系统的Ca2+释放不同。