Sansom S C, Mougouris T, Ono S, DuBose T D
Department of Medicine, University of Texas, Houston 77030.
Am J Physiol. 1994 Sep;267(3 Pt 2):F489-96. doi: 10.1152/ajprenal.1994.267.3.F489.
The inner medullary collecting duct (IMCD) in vivo has the capacity to either secrete or reabsorb K+. However, a selective K+ conductance has not been described previously in the IMCD. In the present study, the patch-clamp method was used to determine the presence and properties of K(+)-selective channels in the apical membrane of the inner medullary collecting duct cell line, mIMCD-3. Two types of K(+)-selective channels were observed in both cell-attached and excised patches. The most predominant K+ channel, a smaller conductance K+ channel (SK), was present in cell-attached patches with 140 mM KCl (high bath K+) but not with 135 mM NaCl plus 5 mM KCl (low bath K+) in the bathing solution. The single-channel conductance of SK was 36 pS with inward currents and 29 pS with outward currents in symmetrical 140 mM KCl. SK was insensitive to both voltage and Ca2+. However, SK was inhibited significantly by millimolar concentrations of ATP in excised patches. A second K(+)-selective channel [a larger K+ channel (BK)] displayed a single-channel conductance equal to 132 pS with inward currents and 90 pS with outward currents in symmetrical 140 mM KCl solutions. BK was intermittently activated in excised inside-out patches by Mg(2+)-ATP in concentrations from 1 to 5 mM. With complete removal of Mg2+, BK was insensitive to ATP. BK was also insensitive to potential and Ca2+ and was observed in cell-attached patches with 140 mM KCl in the bath solution. Both channels were blocked reversibly by 1 mM Ba2+ from the intracellular surface but not by external Ba2+.(ABSTRACT TRUNCATED AT 250 WORDS)
体内的内髓集合管(IMCD)具有分泌或重吸收钾离子(K⁺)的能力。然而,此前尚未在内髓集合管中描述过选择性钾离子电导。在本研究中,采用膜片钳方法来确定内髓集合管细胞系mIMCD - 3顶端膜中钾离子选择性通道的存在及其特性。在细胞贴附式和游离膜片上均观察到两种类型的钾离子选择性通道。最主要的钾离子通道,即较小电导的钾离子通道(SK),存在于细胞贴附式膜片中,此时胞外溶液为含140 mM KCl(高胞外钾离子浓度),而非含135 mM NaCl加5 mM KCl(低胞外钾离子浓度)的溶液。在对称的140 mM KCl溶液中,SK的单通道电导内向电流为36 pS,外向电流为29 pS。SK对电压和钙离子(Ca²⁺)均不敏感。然而,在游离膜片中,毫摩尔浓度的三磷酸腺苷(ATP)能显著抑制SK。第二种钾离子选择性通道[较大的钾离子通道(BK)]在对称的140 mM KCl溶液中,内向电流的单通道电导为132 pS,外向电流为90 pS。在游离的内面向外膜片中,1至5 mM浓度的镁离子 - 三磷酸腺苷(Mg(2 +)-ATP)能间歇性激活BK。完全去除镁离子后,BK对ATP不敏感。BK对电位和钙离子也不敏感,在胞外溶液含140 mM KCl的细胞贴附式膜片中也能观察到。两种通道从细胞内表面均可被1 mM钡离子(Ba²⁺)可逆性阻断,但细胞外钡离子无此作用。(摘要截取自250字)