Osmotic shrinkage activates nonselective cation (NSC) channels in various cell types

被引:52
作者
Koch, JP
Korbmacher, C
机构
[1] Univ Oxford, Physiol Lab, Oxford OX1 3PT, England
[2] Goethe Univ Frankfurt, Zentrum Physiol, D-60590 Frankfurt, Germany
基金
英国惠康基金;
关键词
cell shrinkage; volume regulation; patch clamp; cell lines; flufenamate; cation channels;
D O I
10.1007/s002329900503
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Osmotic cell shrinkage activates a nonselective cation (NSC) channel in M-l mouse cortical collecting duct cells (Volk, Fromter & Korbmacher, 1995, Proc. Natl. Acad. Sci. USA 92: 8478-8482). To see whether shrinkage-activated NSC channels are an ubiquitous phenomenon, we tested the effect of hypertonic extracellular solution on whole-cell currents of HT29 human colon carcinoma cells, BSC-1 renal epithelial cells, A10 vascular smooth muscle cells, and Neuro-a neuroblastoma cells. Addition of 100 mM sucrose to an isotonic NaCl bath solution induced cell shrinkage of HT29 cells as evidenced by a decrease in cell diameter from 18 +/- 1 mu m to 12 +/- 1 mu m (n = 13). Upon cell shrinkage whole-eel currents of HT29 cells increased within 8 +/- 1 min by about 30-fold(n = 13). Cell shrinkage and current activation were reversible upon return to isotonic solution. Replacement of bath Na+ by K+ or Li+ had almost no effect on the stimulated inward current. In contrast, replacement by N-methyl-D-glucamine (NMDG) completely abolished it and shifted the reversal potential from -4.5 +/- 0.7 mV to -57 +/- 4.1 mV (12 = 10). Thus, the stimulated conductance is nonselective for alkali cations bur highly selective for cations over anions with a cation-to-anion permeability ratio of about 13. Flufenamic acid (100 mu M) inhibited the stimulated current by 84 +/- 4.7% (n = 8). During the early phase of hypertonic stimulation single-channel transitions could be detected in whole-cell current recordings, and a gradual activation of 12 and more individual channels with a single-channel conductance of 17.6 +/- 0.9 pS (Iz = 4) could be resolved. In analogous experiments similar shrinkage-activated NSC channels were also observed in BSC-1 renal epithelial cells, A10 vascular smooth muscle cells, and Neuro-2a neuroblastoma cells. These findings indicate that shrinkage-activated NSC channels are an ubiquitous phenomenon and may play a role in volume regulation.
引用
收藏
页码:131 / 139
页数:9
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