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Am J Physiol Regul Integr Comp Physiol 294: R1033-R1043, 2008. First published January 9, 2008; doi:10.1152/ajpregu.00516.2007
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COMPARATIVE AND EVOLUTIONARY PHYSIOLOGY

Regulation of ion transport by pH and [HCO3] in isolated gills of the crab Neohelice (Chasmagnathus) granulata

Martin Tresguerres,1,2,* Scott K. Parks,1,* Sebastian E. Sabatini,3,4 Greg G. Goss,1 and Carlos M. Luquet3,5

1Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada; 2Department of Pharmacology, Weill Medical College of Cornell University, New York, New York; 3Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina; 4Departamento Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina; and 5Asentamiento Universitario San Martín de los Andes, Neuquén, Argentina

Submitted 16 July 2007 ; accepted in final form 7 January 2008

Posterior isolated gills of Neohelice (Chasmagnathus) granulatus were symmetrically perfused with hemolymph-like saline of varying [HCO3] and pH. Elevating [HCO3] in the saline from 2.5 to 12.5 mmol/l (pH 7.75 in both cases) induced a significant increase in the transepithelial potential difference (Vte), a measure of ion transport. The elevation in [HCO3] also induced a switch from acid secretion (–43.7 ± 22.5 µequiv·kg–1·h–1) in controls to base secretion (84.7 ± 14.4 µequiv·kg–1·h–1). The HCO3-induced Vte increase was inhibited by basolateral acetazolamide (200 µmol/l), amiloride (1 mmol/l), and ouabain (5 mmol/l) but not by bafilomycin (100 nmol/l). The Vte response to HCO3 did not take place in Cl-free conditions; however, it was unaffected by apical SITS (2 mmol/l) or DIDS (1 mmol/l). A decrease in pH from 7.75 to 7.45 pH units in the perfusate also induced a significant increase in Vte, which was matched by a net increase in acid secretion of 67.8 ± 18.4 µequiv kg–1 h–1. This stimulation was sensitive to basolateral acetazolamide, bafilomycin, DIDS, and Na+-free conditions, but it still took place in Cl-free saline. Therefore, the cellular response to low pH is different from the HCO3-stimulated response. We also report V-H+-ATPase- and Na+-K+-ATPase-like immunoreactivity in gill sections for the first time in this crab. Our results suggest that carbonic anhydrase (CA), basolateral Na+/H+ exchangers and Na+-K+-ATPase and apical anion exchangers participate in the HCO3-stimulated response, while CA, apical V-H+-ATPase and basolateral HCO3-dependent cotransporters mediate the response to low pH.

acid/base regulation; HCO3 secretion; V-H+-ATPase; carbonic anhydrase; electrogenic Na+/H+ exchanger; amiloride; acetazolamide; bafilomycin



Address for reprint requests and other correspondence: M. Tresguerres, Dept. of Pharmacology, Weill Medical College of Cornell Univ., New York, NY 10021 (e-mail: mat2031{at}med.cornell.edu)







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