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Original Paper

Analysis of Blocker-labeled Channels Reveals the Dependence of Recycling Rates of ENaC on the Total Amount of Recycled Channels

Taruno A.1,2 · Marunaka Y.1

Author affiliations

1Department of Molecular Cell Physiology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto,2Research Fellow of the Japan Society for the Promotion of Science

Corresponding Author

Prof. Yoshinori Marunaka, MD, PhD

Department of Molecular Cell Physiology, Graduate School of Medical Science

Kyoto Prefectural University of Medicine, Kyoto 602-8566 (Japan)

Tel. +81-75-251-5310, Fax +81-75-251-0295

E-Mail marunaka@koto.kpu-m.ac.jp

Related Articles for ""

Cell Physiol Biochem 2010;26:925–934

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Trafficking is one of the primary mechanisms of epithelial Na+ channel (ENaC) regulation. Although it is known that ENaCs are recycled between the apical membrane and the intracellular channel pool, it has been difficult to investigate the recycling of ENaCs; especially endogenously expressed ENaCs. The aim of the present study is to investigate if the recycling rates of ENaCs depend on the total amount of recycled ENaCs. To accomplish this point, we established a novel method to estimate the total amount of recycled ENaCs and the ENaC recycling rates by using a specific blocker (benzamil) of ENaC with a high-affinity for functional label of the channels in recycling. Applying this method, we studied if a decrease in the total amount of ENaCs caused by brefeldin A (5 µg/mL, 1 h) affects respectively the rates of insertion and endocytosis of ENaCs to and from the apical membrane in monolayers of renal epithelial A6 cells. Our observations indicate that: 1) both insertion and endocytosis rates of ENaC increase when the total amount of ENaCs decreases, 2) the increase in the insertion rate is larger than that in the endocytosis rate, and 3) this larger increase in the insertion rate than the endocytosis rate caused by the decrease in the total amount of ENaCs plays an important role in preventing Na+ transport from drastically diminishing due to a decrease in the total amount of ENaCs. The newly established analysis of blocker-labeled ENaCs in the present study provides a useful tool to investigate the recycling of endogenously expressed ENaCs.

© 2010 S. Karger AG, Basel

Article / Publication Details

First-Page Preview
Abstract of Original Paper

Accepted: November 15, 2010
Published online: January 04, 2011
Issue release date: January 2011

Number of Print Pages: 10
Number of Figures: 0
Number of Tables: 0

ISSN: 1015-8987 (Print)
eISSN: 1421-9778 (Online)

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