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First published online July 14, 2008
Journal of Experimental Biology 211, 2467-2477 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.017491

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Branchial expression and localization of SLC9A2 and SLC9A3 sodium/hydrogen exchangers and their possible role in acid–base regulation in freshwater rainbow trout (Oncorhynchus mykiss)

G. Ivanis, A. J. Esbaugh and S. F. Perry^*

Department of Biology, University of Ottawa, 30 Marie Curie, Ottawa, Ontario, Canada K1N 6N5

^* Author for correspondence (e-mail: sfperry{at}uottawa.ca)

Accepted 31 March 2008

Experiments were conducted on adult rainbow trout (Oncorhynchus mykiss) to test the hypothesis that SLC9 Na⁺/H⁺ exchangers (SLC9A2, NHE2; and SLC9A3, NHE3) on the gill epithelium are localized specifically to a subset of mitochondria-rich cells (MRCs) that are unable to bind peanut lectin agglutinin (PNA). This cell type, termed the PNA^– MRC, is a sub-type of MRC believed to function in Na⁺ uptake and acid excretion. A technique using biotinylated PNA was used to distinguish between the PNA^– and PNA⁺ MRCs on fixed gill sections. In contrast to expectations, both NHE2 (mRNA) and NHE3 (protein) were confined to cells enriched with Na⁺/K⁺-ATPase and capable of binding PNA. Thus, in trout, NHE2 and NHE3 are localized to PNA⁺ MRCs, the cells previously believed to be responsible for Cl^– uptake and base excretion. Levels of mRNA for NHE2, the predominant isoform in the gill, were increased during 72 h of hypercapnic acidosis; NHE3 mRNA and protein levels were unaffected. Because plasma cortisol levels were increased during hypercapnia (from 35.3±9.4 to 100.1±30.9 ng ml^–1), the effects of experimentally elevated cortisol levels on NHE expression were investigated. The elevation of plasma cortisol using intraperitoneal implants caused a significant increase in NHE2 mRNA expression without affecting NHE3 mRNA or protein abundance. Thus, we suggest that NHE2 contributes to acid–base regulation during hypercapnia owing to its transcriptional regulation by cortisol. The finding of NHE expression in PNA⁺ MRCs is discussed with reference to current models of ionic and acid–base regulation in teleost fish.

Key words: Mitochondria-rich cell, chloride cell, peanut lectin agglutinin, hypercapnia, cortisol, gill, NHE, NHE2, NHE3, sodium/proton exchangers, fish

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