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

  • Ion Channels, Transporters
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Abstract

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, C2-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.

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Acknowledgements

This work was supported by grants from National Institute of Health (NS42236, NS45155) and American Heart Association-Bugher Foundation (0170063N, 0170064N).

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  1. Department of Pharmaceutical Sciences, Medical University of South Carolina, 280 Calhoun Street, P.O. Box 250144, Charleston, SC 29425, USA

    Chun Jin, Aizhen Yang & Shan Ping Yu

  2. Department of Pathology, Medical University of South Carolina, Charleston, SC 29425, USA

    Ling Wei, Zhong Yang Lu & Shan Ping Yu

  3. Department of Neurology, School of Medicine, Washington University, St. Louis, MO 63110, USA

    Ling Wei, Ai Ying Xiao, Aizhen Yang & Shan Ping Yu

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  1. Ling Wei
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Wei, L., Xiao, A.Y., Jin, C. et al. Effects of chloride and potassium channel blockers on apoptotic cell shrinkage and apoptosis in cortical neurons. Pflugers Arch - Eur J Physiol 448, 325–334 (2004). https://doi.org/10.1007/s00424-004-1277-2

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