Gögelein H, Greger R
Pflugers Arch. 1986;407 Suppl 2:S142-8. doi: 10.1007/BF00584943.
The patch-clamp method was applied to the lateral membrane of late proximal tubules of the rabbit kidney. Tubule segments were cannulated on one side by a perfusion system. At the noncannulated end of the tubules, the lateral membrane was accessible to a patch pipette. In cell-attached, as well as cell-excised (presumably inside-out oriented) membrane patches, a voltage sensitive channel was observed. The open-state probability of this channel increased with depolarizing potentials. In cell-excised patches bathed with NaCl-Ringer on both sides, the single channel conductance g was 28.0 +/- 1.2 pS (n = 10). With KCl-Ringer in the pipette and NaCl-Ringer in the bath g was 24.7 +/- 1.3 pS (n = 7) and the current-voltage curve crossed the axis at 0 mV. Therefore, the channel does not discriminate between K+ and Na+ ions. Replacing half of NaCl by mannitol on the bath side yielded a permeability for cations about twice as high as for Cl-. The channel could be reversibly blocked by diphenylamine-2-carboxylate (DPC), whereas its inhibition by SITS was only partially reversible. In cell-attached patches, the channel was nearly inactivated at zero clamp potential, but became active when the membrane patch was depolarized. The significance of this nonselective channel for proximal tubule cell function is still unclear. It could be involved in the contraluminal exit mechanism of various anions. However, it could also play a role in cell volume regulation processes.
膜片钳方法应用于兔肾近端小管晚期的外侧膜。小管节段一侧通过灌注系统插管。在小管的非插管端,外侧膜可被膜片吸管触及。在细胞贴附式以及细胞切除式(推测为内面向外取向)膜片中,观察到一个电压敏感通道。该通道的开放概率随去极化电位增加。在两侧均用NaCl - 林格液灌注的细胞切除式膜片中,单通道电导g为28.0±1.2 pS(n = 10)。当吸管中为KCl - 林格液而浴槽中为NaCl - 林格液时,g为24.7±1.3 pS(n = 7),电流 - 电压曲线在0 mV处与轴相交。因此,该通道对K⁺和Na⁺离子无选择性。在浴槽侧用甘露醇替代一半的NaCl,阳离子通透性约为Cl⁻的两倍。该通道可被二苯胺 - 2 - 羧酸盐(DPC)可逆性阻断,而其被SITS抑制仅部分可逆。在细胞贴附式膜片中,该通道在零钳制电位时几乎失活,但当膜片去极化时变得活跃。这种非选择性通道对近端小管细胞功能的意义仍不清楚。它可能参与各种阴离子的对腔输出机制。然而,它也可能在细胞体积调节过程中起作用。