Kawakubo T, Naruse K, Matsubara T, Hotta N, Sokabe M
Department of Internal Medicine III, Nagoya University School of Medicine, Nagoya 466-8550, Japan.
Am J Physiol. 1999 Jun;276(6):H1827-38. doi: 10.1152/ajpheart.1999.276.6.H1827.
With the use of the patch-clamp technique, five kinds of stretch-activated (SA) ion channels were identified on the basis of their single-channel conductances and ion selectivities in cultured chick ventricular myocytes. Because a high-conductance K+-selective channel predominated among these channels, we concentrated on characterizing its properties mostly using excised inside-out patches. With 145 mM KCl solution in the pipette and the bath, the channel had a conductance of 199.8 +/- 8.2 pS (n = 22). The ion selectivities among K+, Na+, Ca2+, and Cl- as estimated from their permeability ratios were PNa/PK = 0.03, PCa/PK = 0.025, and PCl/PK = 0.026. The probability of the channel being open (Po) increased with the Ca2+ concentration in the bath ([Ca2+]b; dissociation constant Kd = 0.51 microM at +30 mV) and membrane potential (voltage at half-maximal Po = 39.4 mV at 0.35 microM [Ca2+]b). The channel was blocked by gadolinium, tetraethylammonium, and charybdotoxin from the extracellular surface and, consequently, was identified as a Ca2+-activated K+ (KCa) channel type. The channel was also reversibly activated by ATP applied to the intracellular surface (Kd = 0.74 mM at 0.10 microM [Ca2+]b at +30 mV). From these data taken together, we concluded that the channel is a new type of KCa channel that could be designated as an "SA KCa,ATP channel." To our knowledge, this is the first report of KCa channel in heart cells.
利用膜片钳技术,根据培养的鸡心室肌细胞中五种拉伸激活(SA)离子通道的单通道电导和离子选择性对其进行了鉴定。由于在这些通道中高电导钾离子选择性通道占主导地位,我们主要使用切除的内面向外膜片来集中研究其特性。当移液管和浴槽中均为145 mM KCl溶液时,该通道的电导为199.8±8.2 pS(n = 22)。根据钾离子、钠离子、钙离子和氯离子的渗透率比值估算,它们之间的离子选择性为PNa/PK = 0.03、PCa/PK = 0.025和PCl/PK = 0.026。通道开放概率(Po)随浴槽中钙离子浓度([Ca2+]b;在+30 mV时解离常数Kd = 0.51 microM)和膜电位(在0.35 microM [Ca2+]b时Po达到最大值一半时的电压 = 39.4 mV)升高而增加。该通道从细胞外表面被钆、四乙铵和蝎毒素阻断,因此被鉴定为一种钙离子激活钾离子(KCa)通道类型。该通道也可被施加到细胞内表面的ATP可逆激活(在+30 mV、0.10 microM [Ca2+]b时Kd = 0.74 mM)。综合这些数据,我们得出结论,该通道是一种新型的KCa通道,可命名为“SA KCa,ATP通道”。据我们所知,这是关于心脏细胞中KCa通道的首次报道。