Functional characterization and cell immunolocalization of AQP-CD water channel in kidney collecting duct

K. Fushimi, S. Sasaki, T. Yamamoto, M. Hayashi, T. Furukawa, S. Uchida, M. Kuwahara, K. Ishibashi, M. Kawasaki, I. Kihara, F. Marumo

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Abstract

Vasopressin-regulated water permeability of the kidney collecting duct is a key component of the urine concentration machinery. Recently, a cDNA for AQP-CD, the vasopressin-regulated water channel, initially reported as WCH- CD has been isolated (K. Fushimi, S. Uchida, Y. Hara, Y. Hirata, F. Marumo, and S. Sasaki, Nature Lond. 361:549-552, 1993). AQP-CD was expressed in oocyte membrane using a Xenopus expression vector, and functional characteristics of AQP-CD were examined. Osmotic water permeability (P(f)) of oocytes expressing AQP-CD was 138 ± 19 μm/s (mean ± SE), 12 times greater than the control (11 ± 3 μm/s), 90% inhibited by 0.3 mM HgCl2, and weakly temperature dependent (energy of activation for P(f) was 4.0 kcal/mol). Urea influx measured from 15-min [14C]urea uptake by oocytes injected with AQP- CD/expression vector 1 cRNA was 86 ± 17% of the control. Two-electrode voltage-clamp experiments revealed insignificant ion conductance of AQP-CD. Immunoblots of membranes from rat kidney medulla and oocytes expressing AQP- CD using anti-AQP-CD COOH-terminal antibody showed a 29-kDa protein and 35- to 50-kDa high-molecular-mass forms. Immunohistochemistry showed apical and subapical localization of AQP-CD in the collecting duct principal cells. Our results indicated that AQP-CD is a 29-kDa protein, a selective water channel, distinct from a urea channel, and localized to the membranes of vasopressin- sensitive components in kidney collecting duct principal cells.

Original languageEnglish
Pages (from-to)F573-F582
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume267
Issue number4 36-4
DOIs
Publication statusPublished - 1994
Externally publishedYes

Keywords

  • osmotic water permeability
  • urine concentration
  • vasopressin
  • water channel of collecting duct

ASJC Scopus subject areas

  • Physiology

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    Fushimi, K., Sasaki, S., Yamamoto, T., Hayashi, M., Furukawa, T., Uchida, S., Kuwahara, M., Ishibashi, K., Kawasaki, M., Kihara, I., & Marumo, F. (1994). Functional characterization and cell immunolocalization of AQP-CD water channel in kidney collecting duct. American Journal of Physiology - Renal Fluid and Electrolyte Physiology, 267(4 36-4), F573-F582. https://doi.org/10.1152/ajprenal.1994.267.4.f573