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First published online October 21, 2004
Journal of Experimental Biology 207, vi (2004)
Copyright © 2004 The Company of Biologists Limited
doi: 10.1242/jeb.01301
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Outside JEB

NEW ATPASE FOUND IN ATLANTIC STINGRAY

M. Danielle McDonald

University of Miami

dmcdonald{at}rsmas.miami.edu


An exciting new discovery has been made in the Atlantic stingray that has implications for ion regulation in elasmobranchs. In mammals, an active transport protein (HK{alpha}1) is present in the stomach and kidney that mediates proton (H+ or acid) secretion in exchange for potassium ions (K+) and functions in digestion, acid–base balance and ion regulation. Like mammals, elasmobranchs (sharks, skates and rays) are also believed to secrete acid into their stomachs, but unlike mammals, they primarily use their gills for acid excretion and ion regulation instead of their kidneys. Keith Choe, working with David Evans and his team, set out to determine whether an HK{alpha}1-like protein exists in the Atlantic stingray and to determine if it is expressed in the gill.

The group searched for HK{alpha}1 in the Atlantic stingray gill and stomach by using an antibody made against pig HK{alpha}1. With this antibody, the group identified HK{alpha}1 in gill cells rich in Na+/K+ ATPase. These cells are important in acid secretion and ion absorption in fish. HK{alpha}1 immunoreactivity was not found in cells that stained for V-type H+-ATPase; cells that are thought to be important in base secretion in fish. Knowing that HK{alpha}1 is found in mammalian gastric glands, the team also found HK{alpha}1 immunoreactivity in the stomach of the stingray. Having found that the stingray produces HK{alpha}1, the next step was to determine whether its expression is regulated during freshwater exposure or ion regulation.

Characterising stingray HK{alpha}1 on a molecular level, Evan's team found that it is highly similar to rat HK{alpha}1, and that it is expressed in the stingray's stomach and gill but, surprisingly, not in the kidney. Using quantitative, real-time PCR, the team determined that the expression of gill HK{alpha}1 does not change with elevated environmental CO2 but is higher in fish acclimated to freshwater than those acclimated to seawater suggesting that it plays a role in freshwater acclimation.

That the expression of HK{alpha}1 in the gill is unaffected by the respiratory acidosis caused by high environmental CO2 suggests that this gene is not involved in acid–base balance. However, this finding does not exclude the involvement of pre-existing gill HK{alpha}1 proteins in response to acid–base disturbance. Importantly, HK{alpha}1 is upregulated in freshwater-acclimated stingrays, suggesting that it may act as a mechanism to increase active K+ uptake across the gill when environmental K+ levels are low, possibly making HK{alpha}1 vital for the physiological fitness of this freshwater organism.

This study is the first to provide direct evidence for the presence of HK{alpha}1 in any fish. From an evolutionary perspective, stomach acid secretion first appeared in elasmobranchs, which suggests that the Atlantic stingray may be one of the oldest living organisms to have HK{alpha}1.

References

Choe, K. P, Verlander, J. W., Wingo, C. S., and Evans, D. H. (2004). A putative H+/K+ ATPase in the Atlantic stingray, Dasyatis sabina: primary sequence and expression in gills. Am. J. Physiol. Regul. Integr. Comp. Physiol. 10.1152/ajpregu.00513.2003[Abstract/Free Full Text]





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