S. M. RAHIM, S. J. ABDURRAHMAN, E. M. TAHA, H. F. HASSAN, K. D. SIMON, A. G. MAZLAN

Immunohistochemical and partial characterization studies of carbonic anhydrase in the brain of teleost fish Cyprinus carpio

Carbonic anhydrase (CA) activity is related to a variety of intermingled biological processes, including respiration, ionic transport, acid–base balance, and calcification. Carps Cyprinus carpio were investigated with respect to purification, partial characterization, and immunohistochemical localization of brain CA. The enzyme was purified up to 72 fold by affinity chromatography on sulfanilamide sepharose gel. The result is a yield of 59.6% and a specific activity of 1143 EU/mg protein with an apparent molecular weight of 28.6 kDa. The optimum pH and temperature were observed at 8.0 and 35 oC, respectively. Ions of Na+, K+, Mg2+, and Ca2+ at concentrations of 1, 5, and 10 mM exhibited up to 25% to 30% inhibition on brain CA activity, whereas Cl– exhibited the most inhibition on enzyme activity. Acetazolamide also displayed significant inhibition on brain CA activity (54%, 83%, and 92% at respective concentrations of 1, 5, and 10 mM). Antibodies against purified gill CA from trout Oncorhynchus mykiss reacted positively only to one band corresponding to CA in carp brain supernatant. Immunohistochemical staining was observed in oligodendrocytes, its processes, and in myelinated fibers. Our findings are similar to those of previous studies on higher vertebrates CA II.

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