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Table of Contents
Vol. 21, No. 1-3, 2008
Issue release date: January 2008
Section title: Original Paper
Cell Physiol Biochem 2008;21:183–192
(DOI:10.1159/000113760)

TRPC6 Contributes to the Ca2+ Leak of Human Erythrocytes

Föller M.* · Kasinathan R.S.* · Koka S.* · Lang C.* · Shumilina E.* · Birnbaumer L.1 · Lang F.1 · Huber S.M.1
Department of Physiology, University of Tübingen,1Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC,*M.F. and R.S.K. contributed equally to this study and therefore share first-authorship
email Corresponding Author

Stephan Huber

Department of Physiology I, Eberhard-Karls-University Tübingen

Gmelinstrasse 5, 72076 Tübingen (Germany)

Tel. +49 7071-29-76737, Fax: +49 7071-29-3073

E-Mail stephan.huber@uni-tuebingen.de

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Abstract

Human erythrocytes express cation channels which contribute to the background leak of Ca2+, Na+ and K+. Excessive activation of these channels upon energy depletion, osmotic shock, Cl- depletion, or oxidative stress triggers suicidal death of erythrocytes (eryptosis), characterized by cell-shrinkage and exposure of phosphatidylserine at the cell surface. Eryptotic cells are supposed to be cleared from circulating blood. The present study aimed to identify the cation channels. RT-PCR revealed mRNA encoding the non-selective cation channel TRPC6 in erythroid progenitor cells. Western blotting indicated expression of TRPC6 protein in erythrocytes from man and wildtype mice but not from TRPC6-/- mice. According to flow-cytometry, Ca2+ entry into human ghosts prepared by hemolysis in EGTA-buffered solution containing the Ca2+ indicator Fluo3/AM was inhibited by the reducing agent dithiothreitol and the erythrocyte cation channel blockers ethylisopropylamiloride and amiloride. Loading of the ghosts with antibodies against TRPC6 or TRPC3/6/7 but neither with antibodies against TRPM2 or TRPC3 nor antibodies pre-adsorbed with the immunizing peptides inhibited ghost Ca2+ entry. Moreover, free Ca2+ concentration, cell-shrinkage, and phospholipid scrambling were significantly lower in Cl--depleted TRPC6-/- erythrocytes than in wildtype mouse erythrocytes. In conclusion, human and mouse erythrocytes express TRPC6 cation channels which participate in cation leak and Ca2+-induced suicidal death.

© 2008 S. Karger AG, Basel


Article / Publication Details

First-Page Preview
Abstract of Original Paper

Accepted: June 20, 2007
Published online: January 16, 2008
Issue release date: January 2008

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

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

For additional information: http://www.karger.com/CPB


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