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Distinct pharmacological and molecular properties of the acid-sensitive outwardly rectifying (ASOR) anion channel from those of the volume-sensitive outwardly rectifying (VSOR) anion channel

  • Ion channels, receptors and transporters
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Abstract

Expressed by many cell types, acid-sensitive outwardly rectifying (ASOR) anion channels are known to be activated by extracellular acidification and involved in acidotoxic necrotic cell death. In contrast, ubiquitously expressed volume-sensitive outwardly rectifying (VSOR) anion channels are activated by osmotic cell swelling and involved in cell volume regulation and apoptotic cell death. Distinct inhibitors to distinguish ASOR from VSOR anion channels have not been identified. Although leucine-rich repeats containing 8A (LRRC8A) was recently found to be an essential component of VSOR anion channels, the possibility of an LRRC8 family member serving as a component of ASOR anion channels has not been examined. In this study, we explored the effects of 12 known VSOR channel inhibitors and small interfering RNA (siRNA)-mediated knockdown of LRRC8 family members on ASOR and VSOR currents in HeLa cells. Among these inhibitors, eight putative VSOR blockers, including 4-(2-butyl-6,7-dichlor-2-cyclopentylindan-1-on-5-yl) oxobutyric acid (DCPIB) and 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), were totally ineffective at blocking ASOR channel activity, whereas suramin, R-(+)-[(2-n-butyl-6,7-dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1H-inden-5-yl)oxy] acetic acid (DIOA), arachidonic acid, and niflumic acid were found to be effective ASOR anion channel antagonists. In addition, gene-silencing studies showed that no LRRC8 family members are essentially involved in ASOR anion channel activity, whereas LRRC8A is involved in VSOR anion channel activity in HeLa cells.

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Acknowledgments

This work was supported by the JSPS KAKENHI grants in-aid (Numbers 25670112 and 15K15028 to Y.O. and Number 26860153 to K.S.-N.). The authors thank R. Z. Sabirov for discussion as well as K. Shigemoto and N. Yasui for technical assistance.

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Authors and Affiliations

  1. Japan Society for the Promotion of Science, Chiyoda-ku, Tokyo, 102-0083, Japan

    Kaori Sato-Numata

  2. Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, 817-0180, Japan

    Kaori Sato-Numata, Tomohiro Numata & Ryuji Inoue

  3. National Institute for Physiological Sciences, National Institutes of National Sciences, Okazaki, 444-8787, Japan

    Kaori Sato-Numata & Yasunobu Okada

  4. SOKENDAI (The Graduate University for Advanced Studies), Hayama, Kanagawa, 240-0193, Japan

    Yasunobu Okada

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  1. Kaori Sato-Numata
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Correspondence to Yasunobu Okada.

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Sato-Numata, K., Numata, T., Inoue, R. et al. Distinct pharmacological and molecular properties of the acid-sensitive outwardly rectifying (ASOR) anion channel from those of the volume-sensitive outwardly rectifying (VSOR) anion channel. Pflugers Arch - Eur J Physiol 468, 795–803 (2016). https://doi.org/10.1007/s00424-015-1786-1

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