Effects of chloride and potassium channel blockers on apoptotic cell shrinkage and apoptosis in cortical neurons

被引:98
作者
Wei, L
Xiao, AY
Jin, C
Yang, AZ
Lu, ZY
Yu, SP
机构
[1] Med Univ S Carolina, Dept Pharmaceut Sci, Charleston, SC 29425 USA
[2] Med Univ S Carolina, Dept Pathol, Charleston, SC 29425 USA
[3] Washington Univ, Sch Med, Dept Neurol, St Louis, MO 63110 USA
来源
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY | 2004年 / 448卷 / 03期
关键词
DIDS; volume regulation; Cl-; efflux; K+ efflux; caspase; DNA laddering;
D O I
10.1007/s00424-004-1277-2
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
K+ and Cl- homeostasis have been implicated in cell volume regulation and apoptosis. We addressed the hypothesis that K+ and Cl- efflux may contribute to apoptotic cell shrinkage and apoptotic death in cultured cortical neurons. CLC-2 and CLC-3 chloride channels were detected in cultured cortical neurons. The Cl- channel blockers 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS) and 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) inhibited the outwardly rectifying Cl- current, prevented apoptotic cell shrinkage, and mildly attenuated cell death induced by staurosporine, C-2-ceramide, or serum deprivation. Cl- channel blockers, however, at concentrations that prevented cell shrinkage had no significant effects on caspase activation and/or DNA fragmentation. Cell death in the presence of a Cl- channel blocker was still sensitive to blockade by the caspase inhibitor z-Val-Ala-Asp(OMe)-fluoromethyl ketone (z-VAD-fmk). Electron microscopy revealed that, although DIDS prevented apoptotic cell shrinkage, certain apoptotic ultrastructural alterations still took place in injured neurons. On the other hand, the K+ channel blocker tetraethylammonium (TEA), clofilium, or the caspase inhibitor z-VAD-fmk prevented cell shrinkage as well as caspase activation and/or DNA damage, and showed stronger neuroprotection against apoptotic alterations and cell death. The results indicate that neurons may undergo apoptotic process without cell shrinkage and imply distinct roles for Cl- and K+ homeostasis in regulating different apoptotic events.
引用
收藏
页码:325 / 334
页数:10
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