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Turkish Journal of Fisheries and Aquatic Sciences 2013, Vol 13, Num, 2     (Pages: 341-347)

Immunohistochemical and Partial Characterization Studies of Carbonic Anhydrase in the Brain of Teleost Fish Cyprinus carpio

S. M. Rahim 1-5 ,S. J. Abdurrahman 2 ,E. M. Taha 3 ,H. F. Hassan 4 ,K. D. Simon 5 ,A. G. Mazlan 5

1 Dept. of Biology, Faculty of Education, University of Tikrit, 34001 Tikrit, Salah Al Deen, Iraq
2 Dept. of Biology, Faculty of Science, University of Tikrit, 34001 Tikrit, Salah Al Deen, Iraq
3 Dept. of Chemistry,Faculty of Science for Women,University of Baghdad, Baghdad, Iraq
4 Dept. of Biology, Faculty of Science, University of Kirkuk, Kirkuk, Iraq
5 School of Environmental and Natural Resource Sciences, Faculty of Science and Technology,Universiti Kebangsaan Malaysia,43600 UKM Bangi Selangor, D.E., Malaysia
DOI : 10.4194/1303-2712-v13_2_16 Viewed : 4200 - Downloaded : 3067 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. Keywords : Teleost fish, enzyme purification, immunohistochemistry, metal ions, oligodendrocytes