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A pure chloride channel mutant of CLC-5 causes Dent’s disease via insufficient V-ATPase activation

  • Ion channels, receptors and transporters
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Abstract

Dent’s disease is characterized by defective endocytosis in renal proximal tubules (PTs) and caused by mutations in the 2Cl−/H+ exchanger, CLC-5. However, the pathological role of endosomal acidification in endocytosis has recently come into question. To clarify the mechanism of pathogenesis for Dent’s disease, we examined the effects of a novel gating glutamate mutation, E211Q, on CLC-5 functions and endosomal acidification. In Xenopus oocytes, wild-type (WT) CLC-5 showed outward-rectifying currents that were inhibited by extracellular acidosis, but E211Q and an artificial pure Cl− channel mutant, E211A, showed linear currents that were insensitive to extracellular acidosis. Moreover, depolarizing pulse trains induced a robust reduction in the surface pH of oocytes expressing WT CLC-5 but not E211Q or E211A, indicating that the E211Q mutant functions as a pure Cl− channel similar to E211A. In HEK293 cells, E211A and E211Q stimulated endosomal acidification and hypotonicity-inducible vacuolar-type H+-ATPase (V-ATPase) activation at the plasma membrane. However, the stimulatory effects of these mutants were reduced compared with WT CLC-5. Furthermore, gene silencing experiments confirmed the functional coupling between V-ATPase and CLC-5 at the plasma membrane of isolated mouse PTs. These results reveal for the first time that the conversion of CLC-5 from a 2Cl−/H+ exchanger into a Cl− channel induces Dent’s disease in humans. In addition, defective endosomal acidification as a result of insufficient V-ATPase activation may still be important in the pathogenesis of Dent’s disease.

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Acknowledgments

This study was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Department of Internal Medicine, Faculty of Medicine, The University of Tokyo Hospital, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Nobuhiko Satoh, Hideomi Yamada, Masashi Suzuki, Motonobu Nakamura, Atsushi Suzuki & Shoko Horita

  2. Apheresis and Dialysis Center, General Medicine, School of Medicine, Keio University, Tokyo, Japan

    Osamu Yamazaki

  3. Department of Pediatrics, Osaka Medical College, Takatsuki, Osaka, Japan

    Akira Ashida

  4. Biomedical Computation Center, Osaka Medical College, Takatsuki, Osaka, Japan

    Daisuke Yamamoto

  5. Department of Nephrology, Fukuoka Children’s Hospital, Fukuoka, Japan

    Yoshitsugu Kaku

  6. Department of Pediatrics, Ohashi Medical Center, Toho University, Meguro-ku, Tokyo, Japan

    Takashi Sekine

  7. Yaizu City Hospital, Yaizu, Japan

    George Seki

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  1. Nobuhiko Satoh
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  2. Hideomi Yamada
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Correspondence to Shoko Horita.

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Additional information

Daisuke Yamamoto deceased.

Parts of this paper were taken from the thesis written in English by Nobuhiko Satoh. The title of the thesis, which is in Japanese, is as follows: “CLC-5の2Cl/H交換輸送機能はV-ATPaseを介する効率的エンドゾーム酸性化に必要である”. The summary (in Japanese) of the thesis is accessible at http://repository.dl.itc.u-tokyo.ac.jp/index_e.html.

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Satoh, N., Yamada, H., Yamazaki, O. et al. A pure chloride channel mutant of CLC-5 causes Dent’s disease via insufficient V-ATPase activation. Pflugers Arch - Eur J Physiol 468, 1183–1196 (2016). https://doi.org/10.1007/s00424-016-1808-7

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