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Carbonic anhydrase activity in tissues of the icefish Chionodraco hamatus and of the red-blooded teleosts Trematomus bernacchii and Anguilla anguilla

M. Maffia^*, A. Rizzello, R. Acierno, M. Rollo, R. Chiloiro and C. Storelli

Laboratory of General Physiology, Department of Biology, University of Lecce, via Prov.le Monteroni, 73100, Lecce, Italy

View larger version (26K): [in a new window]   Fig. 1. Carbonic anhydrase specific activity in gills (A), intestine (B), kidney (C) and blood (D) of C. hamatus, T. bernacchii and A. anguilla. Values are expressed as means ± S.E.M. of 8 experiments. **P<0.01, *P<0.05 by ANOVA. aDifference between C. hamatus and T. bernacchii; bdifferences between C. hamatus and A. anguilla; cdifferences between T. bernacchii and A. anguilla.

View larger version (14K): [in a new window]   Fig. 2. Carbonic anhydrase activity purified from gills of C. hamatus and A. anguilla, measured at the environmental temperatures of the two species. Values for CO2 trapped (cts min–1) are means ± S.E.M. of 4 experiments. All reported values were significantly different (**P<0.01, ANOVA).

View larger version (17K): [in a new window]   Fig. 3. Inhibitory effect of C. hamatus plasma on the activity of different CA isoforms. CA activity was measured as described in Materials and methods. Values are means ± S.E.M. for 3 different determinations.

View larger version (20K): [in a new window]   Fig. 4. Distribution of total CA activity in gills, kidney, intestine and blood of C. hamatus, T. bernacchii and A. anguilla. CA activity was expressed both as total activity and as the percentage of the sum total of enzymatic activity measured in all of the tissues (see Results). Values are means ± S.E.M. of 8 experiments.

View larger version (39K): [in a new window]   Fig. 5. SDS-PAGE of CA isoforms from C. hamatus, T. bernacchii and A. anguilla gills. The isoforms each migrated as single protein bands of 28.9, 29.9 and 31.2 kDa respectively, as determined from 5 different experiments. The positions of molecular marker proteins are shown.

View larger version (17K): [in a new window]   Fig. 6. Dependence of CA activity on increasing CO2 concentrations, as fitted to a Michaelis–Menten equation. Values are means ± S.E.M. of 3 determinations.

View larger version (16K): [in a new window]   Fig. 7. Effects of sulphanilamide (A) and acetazolamide (ACTZ) (B) on icefish CA activity. (Ai,Bi) Dose–response curves; (Aii,Bii) Easson–Stedman plots. Values are means ± S.E.M. of 3 determinations. For details, see Materials and methods.

View larger version (17K): [in a new window]   Fig. 8. Effect of preincubation temperature (15 min) on the activity of branchial CA isoforms of C. hamatus, T. bernacchii and A. anguilla measured by the electrometric method at 0°C. Values are means ± S.E.M. of 3 experiments. For details, see Materials and methods. The dashed lines indicate the temperatures above which the activity of each isoform is markedly reduced.

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