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Originally published In Press as doi:10.1074/jbc.M004114200 on June 6, 2000

J. Biol. Chem., Vol. 275, Issue 37, 28519-28525, September 15, 2000
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Mammalian ASIC2a and ASIC3 Subunits Co-assemble into Heteromeric Proton-gated Channels Sensitive to Gd3+*

Kazimierz BabinskiDagger §, Stefano CatarsiDagger , Giuseppe Biagini||, and Philippe SéguélaDagger **

From the Dagger  Cell Biology of Excitable Tissue Group, Montreal Neurological Institute, McGill University, Montreal, Quebec, Canada H3A 2B4, § Antalium Inc., Montreal, Quebec, Canada H3A 1X6, and the || Department of Biomedical Sciences, Section of Physiology, University of Modena and Reggio Emilia, 41100 Modena, Italy

Proton receptors of the acid-sensing ion channel (ASIC) family are expressed in sensory neurons and thus could play a critical role in the detection of noxious acidosis. To investigate the subunit composition of native ASICs in peripheral and central neurons, we co-injected human as well as rodent ASIC2a and ASIC3 subunits in Xenopus oocytes. The amplitudes of acid-induced biphasic responses mediated by co-expressed ASIC2a and ASIC3 subunits were much larger (as much as 20-fold) than the currents mediated by the respective homomers, clearly indicating functional association. The reversal potential of the ASIC2a+3 current (>= +20 mV) reflected a cationic current mainly selective for sodium. The sensitivity to pH or amiloride of single versus co-expressed ASIC subunits was not significantly different; however, gadolinium ions inhibited ASIC3 and ASIC2a+3 responses with much higher potency (IC50 ~40 µM) than the ASIC2a response (IC50 >= 1 mM). Biochemical interaction between ASIC2a and ASIC3 subunits was demonstrated by co-purification from transfected human embryonic kidney (HEK293) cells and Xenopus oocytes. Our in situ hybridization data showed that rat ASIC2a and ASIC3 transcripts are co-localized centrally, whereas reverse transcription-polymerase chain reaction data led us to detect co-expression of human ASIC2a and ASIC3 subunits in trigeminal sensory ganglia, brain, and testis where they might co-assemble into a novel subtype of proton-gated channels sensitive to gadolinium.

* This work was supported in part by the Medical Research Council of Canada, Antalium Inc., T2C2-Bio and la Fondation des Maladies du Coeur du Québec.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

These authors contributed equally to this work.

** Senior Scholar of the Fonds de la Recherche en Santé du Québec. To whom correspondence should be addressed: Montreal Neurological Inst., 3801 University St., Rm. 778, Montreal, Quebec, Canada H3A 2B4. Tel.: 1-514-398-5029; Fax: 1-514-398-8106; E-mail: mips@musica.mcgill.ca.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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