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Comparative physiology and molecular analysis of carbonic anhydrase from the red blood cells of teleost fish

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Abstract

This study investigates the early evolution of vertebrate red blood cell (rbc) carbonic anhydrase (CA) by examining the physiological and molecular properties of rbc CA in teleost fish. When representatives of four different families of teleosts were compared, it was found that differences in overall rbc CA activity were due to different concentrations of CA, rather than differences in the enzyme’s kinetic properties. Additional molecular analysis of CA from the rbcs of rainbow trout provided further evidence that critical elements of the enzyme, such as the active site, have been highly conserved during vertebrate evolution. The active site of the trout CA differed from that of gar rbc CA at only two amino acid positions. The rainbow trout rbc CA sequence also showed high sequence homology with CA sequences from other fish tissues, and fits into an emerging group of fish CAs that are basal to mammalian CA I, II and III. Northern blot analysis of the tissue expression of the sequenced CA indicated that it is primarily found in the rbcs, but high amounts of cytosolic CA activity were also found in the gill, suggesting the presence of other cytosolic CA isozymes in this species.

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Abbreviations

Az :

acetazolamide

CA :

carbonic anhydrase

MP :

maximum parsimony

NJ :

neighbour joining

RACE :

rapid amplification of cDNA ends

rbc :

red blood cell

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Acknowledgements

Financial support for this study was provided by a Natural Science and Engineering Council (NSERC) grant to BLT. AJE was supported by Ontario Graduate Studies and NSERC graduate scholarships and SGL was supported by an NSERC graduate scholarship. Special thanks to M. Fortner for technical assistance. These experiments comply with the Canadian Council for Animal Care guidelines and current laws in Canada.

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Authors and Affiliations

  1. Department of Biology, Queen’s University, K7L 3N6, Kingston, Ontario, Canada

    A. J. Esbaugh, S. G. Lund & B. L. Tufts

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  1. A. J. Esbaugh
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  2. S. G. Lund
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  3. B. L. Tufts
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Correspondence to A. J. Esbaugh.

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Communicated by L.C.-H. Wang

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Esbaugh, A.J., Lund, S.G. & Tufts, B.L. Comparative physiology and molecular analysis of carbonic anhydrase from the red blood cells of teleost fish. J Comp Physiol B 174, 429–438 (2004). https://doi.org/10.1007/s00360-004-0430-8

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