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蜥蜴视网膜离体视杆细胞中钠钙钾交换体对钾离子的转运

Transport of K+ by Na(+)-Ca2+, K+ exchanger in isolated rods of lizard retina.

作者信息

Rispoli G, Navangione A, Vellani V

机构信息

INFM, Dipartimento di Biologia dell'Università, Ferrara, Italy.

出版信息

Biophys J. 1995 Jul;69(1):74-83. doi: 10.1016/S0006-3495(95)79877-0.

Abstract

Transport of K+ by the photoreceptor Na(+)-Ca2+, K+ exchanger was investigated in isolated rod outer segments (OS) by recording membrane current under whole-cell voltage-clamp conditions. Known amounts of K+ were imported in the OS through the Ca(2+)-activated K+ channels while perfusing with high extracellular concentration of K+, [K+]o. These channels were detected in the recordings from the OS, which probably retained a small portion of the rest of the cell. The activation of forward exchange (Na+ imported per Ca2+ and K+ extruded) by intracellular K+, Ki+, was described by first-order kinetics with a Michaelis constant, Kapp(Ki+), of about 2 mM and a maximal current, Imax, of about -60 pA. [Na+]i larger than 100 mM had little effect on Kapp(Ki+) and Imax, indicating that Nai+ did not compete with Ki+ for exchange sites under physiological conditions, and that Na+ release at the exchanger intracellular side was not a rate-limiting step for the exchange process. Exchanger stoichiometry resulted in one K+ ion extruded per one positive charge imported. Exchange current was detected only if Ca2+ and K+ were present on the same membrane side, and Na+ was simultaneously present on the opposite side. Nonelectrogenic modes of ion exchange were tested taking advantage of the hindered diffusion found for Cai2+ and Ki+. Experiments were carried out so that the occurrence of a putative nonelectrogenic ion exchange, supposedly induced by the preapplication of certain extracellular ion(s), would have resulted in the transient presence of both Cai2+ and Ki+. The lack of electrogenic forward exchange in a subsequent switch to high Nao+, excluded the presence of previous nonelectrogenic transport.

摘要

通过在全细胞电压钳制条件下记录膜电流,研究了光感受器钠钙钾交换体对钾离子的转运,实验对象为分离的视杆细胞外段(OS)。在细胞外高钾浓度([K+]o)灌注的情况下,已知量的钾离子通过钙激活钾通道进入视杆细胞外段。在视杆细胞外段的记录中检测到了这些通道,视杆细胞外段可能保留了细胞其余部分的一小部分。细胞内钾离子(Ki+)对正向交换(每转运一个钙离子进入同时转运一个钾离子出去)的激活作用,可用一级动力学来描述,其米氏常数Kapp(Ki+)约为2 mM,最大电流Imax约为 -60 pA。细胞内钠离子([Na+]i)浓度大于100 mM时,对Kapp(Ki+)和Imax影响很小,这表明在生理条件下,细胞内钠离子不会与钾离子竞争交换位点,并且交换体胞内侧的钠离子释放不是交换过程的限速步骤。交换体化学计量关系为每转运一个正电荷进入就有一个钾离子被转运出去。只有当钙离子和钾离子在膜的同一侧,且钠离子同时在膜的另一侧时,才能检测到交换电流。利用钙离子和钾离子存在的扩散阻碍,测试了离子交换的非电生模式。实验设计使得在预先施加某些细胞外离子后,假定发生的非电生离子交换会导致钙离子和钾离子同时短暂出现。随后切换到高细胞外钠离子浓度时缺乏电生正向交换,排除了之前存在非电生转运的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8c14/1236226/bc3927589edf/biophysj00059-0076-a.jpg

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