Branchial carbonic anhydrase is present in the dogfish, Squalus acanthias

The distribution of branchial carbonic anhydrase (CA) in the shark, Squalus acanthias, was studied using in situ measurements of pH disequilibrium states in post-branchial saline, and immunological techniques, immunofluorescence microscopy and Western analysis, employing rabbit polyclonal antibodies against rat pulmonary membrane associated CA IV and chick retinal cytosolic CA II. In the in situ saline perfused gill preparation, the CA inhibitor acetazolamide produced a pH disequilibium (0.063 ± 0.022 pH units) while control and bovine carbonic anhydrase perfusions did not (0.012 ± 0.017 and 0.023 ± 0.018 pH units, respectively). These results indicate that the HCO3- dehydration reaction is accelerate by endogenous extracellular CA. Western analysis of saline perfused gill membrane preparations revealed an immunoreactive 48 kDa band with the CA IV probe. In crude gill homogenates, a 33 kDa and 31 kDa pair of bands is identified by the CA II probe. The pattern of immunolabeling for CA II in the gill epithelium was either diffuse or punctate within both lamellar and filament epithelial cells while eyrthrocytes and pillar cells displayed a diffuse staining pattern.