ClC-5: Role in endocytosis in the proximal tubule

Yinghong Wang, Hui Cai, Liudmila Cebotaru, Deanne H. Hryciw, Edward J. Weinman, Mark Donowitz, Sandra E. Guggino, William B. Guggino

Research output: Contribution to journalArticlepeer-review

Abstract

The proper functioning of the Cl- channel, ClC-5, is essential for the uptake of low molecular mass proteins through receptor-mediated endocytosis in the proximal tubule. Dent's disease patients with mutant ClC-5 channels and ClC-5 knockout (KO) mice both have low molecular mass proteinuria. To further understand the function of ClC-5, endocytosis was studied in LLC-PK1 cells and primary cultures of proximal tubule cells from wild-type (WT) and ClC-5 KO kidneys. Endocytosis in the proximal tubule cells from KO mice was reduced compared with that in WT animals. Endocytosis in WT but not in KO cells was inhibited by bafilomycin A-1 and Cl- depletion, whereas endocytosis in both WT and KO cells was inhibited by the NHE3 blocker, S3226. Infection with adenovirus containing WT ClC-5 rescued receptor-mediated endocytosis in KO cells, whereas infection with any of the three disease-causing mutants, myc-W22G-ClC-5, myc-S520P-ClC-5, or myc-R704X-ClC-5, did not. WT and the three mutants all trafficked to the apical surface, as assessed by surface biotinylation. WT-ClC-5 and the W22G mutant were internalized similarly, whereas neither the S520P nor the R704X mutants was. These data indicate that ClC-5 is important for CP and proton pump-mediated endocytosis. However, not all receptor-mediated endocytosis in the proximal tubule is dependent on ClC-5. There is a significant fraction that can be inhibited by an NHE3 blocker. Our data from the mutants suggest that defective targeting and trafficking. of mutant ClC-5 to the endosomes are a major determinant in the lack of normal endocytosis in Dent's disease.

Original languageEnglish (US)
Pages (from-to)F850-F862
JournalAmerican Journal of Physiology - Renal Physiology
Volume289
Issue number4 58-4
DOIs
StatePublished - Oct 2005

Keywords

  • Adenovirus
  • Albumin
  • Bafilomycin A-1
  • Dextran
  • Gluconate
  • Knockout mice
  • Na-H exchanger type 3

ASJC Scopus subject areas

  • Physiology
  • Urology

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