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First published online June 15, 2006
Journal of Experimental Biology 209, 2595-2605 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02286

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Functional evidence for the presence of a carbonic anhydrase repressor in the eyestalk of the euryhaline green crab Carcinus maenas

Raymond P. Henry

Department of Biological Sciences, 101 Life Science Building, Auburn University, Auburn, AL 36849, USA

View larger version (14K): [in a new window]   Fig. 1. Carbonic anhydrase activity (A; µmol CO2 mg protein–1 min–1) and mRNA expression (B; relative amounts) in anterior (G4, black bars) and posterior (G8, gray bars) of green crabs acclimated to 32 p.p.t. salinity vs eyestalk ablated crabs at 32 p.p.t. for 7 days. Values are means ± s.e.m. (N=6–8). T=12°C. Asterisks indicate statistical differences in G8 from the 32 p.p.t. controls

View larger version (12K): [in a new window]   Fig. 2. Carbonic anhydrase activity (µmol CO2 mg protein–1 min–1) in anterior (G4, black bars) and posterior (G8, gray bars) gills of green crabs acclimated to 32 p.p.t. 32, 32 p.p.t. acclimated controls; Sham, intact crabs injected daily with filtered seawater for 7 days; ESA, eyestalk ablated crabs assayed after 7 days; ESA-inj, eyestalk ablated crabs injected daily with eyestalk extract and assayed after 7 days. Values are means ± s.e.m. (N=6–10). T=12°C. Asterisks indicate statistical differences at P<0.05 in G4 and G8 vs the corresponding gill in the 32 p.p.t. controls.

View larger version (11K): [in a new window]   Fig. 3. Carbonic anhydrase activity (A; µmol CO2 mg protein–1 min–1) and mRNA expression (B; relative amounts) in anterior (G4, black bars) and posterior (G8, gray bars) gills of green crabs acclimated to 32 p.p.t. (32) and transferred to 10 p.p.t. for 7 days (10-7days). Values are means ± s.e.m. (N=6). T=12°C. Asterisks indicate statistical differences in G8 from the 32 p.p.t. controls at P<0.05.

View larger version (12K): [in a new window]   Fig. 4. Carbonic anhydrase activity (µmol CO2 mg protein–1 min–1) in anterior (G4, black bars) and posterior (G8, gray bars) gills of green crabs acclimated to 32 p.p.t. and transferred to 10 p.p.t. 32, 32 p.p.t. acclimated controls; 10 intact, intact crabs transferred to 10 p.p.t. for 7 days; 10 sham, intact crabs transferred to 10 p.p.t. and given daily injections of filtered seawater; 10 inj, intact crabs transferred to 10 p.p.t. and given daily injections of eyestalk extract taken from `donor' crabs acclimated to 32 p.p.t. Values are means ± s.e.m. (N=6–9). T=12°C. Asterisks indicate statistical differences in G4 vs 32 p.p.t. controls; different letters indicate statistical differences in G8 across treatments at P<0.05.

View larger version (12K): [in a new window]   Fig. 5. Carbonic anhydrase activity (µmol CO2 mg protein–1 min–1) in anterior (G4, black bars) and posterior (G8, gray bars) gills of green crabs acclimated to 32 p.p.t. and transferred to 10 p.p.t. for 7 days. 32, 32 p.p.t. acclimated controls; 10 intact, intact crabs transferred to 10 p.p.t. for 7 days; 10 ESA, eyestalk ablated crabs transferred to 10 p.p.t. and assayed after 7 days; 10 ESAinj, eyestalk ablated crabs transferred to 10 p.p.t. and given daily injections of eyestalk extract taken from `donor' crabs acclimated to 32 p.p.t. Values are means ± s.e.m. (N=6–9). T=12°C. Different letters indicate statistical differences at P<0.05 in G8 across treatments.

View larger version (12K): [in a new window]   Fig. 6. Carbonic anhydrase activity (A; µmol CO2 mg protein–1 min–1) and mRNA expression (B; relative amounts) in anterior (G4, black bars) and posterior (G8, gray bars) gills of green crabs acclimated to 32 p.p.t. and transferred to 10 p.p.t. for 4 days. Values are means ± s.e.m. (N=6). T=12°C. Asterisks indicate statistical differences in G8 at P<0.05.

View larger version (13K): [in a new window]   Fig. 7. Carbonic anhydrase activity (µmol CO2 mg protein–1 min–1) in anterior (G4, black bars) and posterior (G8, gray bars) of green crabs acclimated to 32 p.p.t. and transferred to 10 p.p.t. for 4 days. 32, 32 p.p.t. acclimated controls; 10-4days, intact crabs assayed after 4 days at 10 p.p.t.; 10-4daysESA, eyestalk ablated crabs assayed after 4 days at 10 p.p.t.. Values are means ± s.e.m. (N=6). T=12°C. Different letters indicate statistical differences in G8 at P<0.05 across treatments.

View larger version (13K): [in a new window]   Fig. 8. Carbonic anhydrase activity (µmol CO2 mg protein–1 min–1) in anterior (G4, black bars) and posterior (G8, gray bars) of green crabs acclimated to 32 p.p.t. and transferred to 10 p.p.t. for 4 days. 32, 32 p.p.t. acclimated controls; 10-4days, intact crabs assayed after 4 days at 10 p.p.t.; 10 inj1x, intact crabs given a once daily injection of eyestalk extract from `donor' crabs acclimated to 32 p.p.t.; 10 inj2x, intact crabs given twice-daily injections of eyestalk extract from `donor' crabs acclimated to 32 p.p.t. Values are means ± s.e.m. (N=6-7). T=12°C. Asterisk indicates statistical difference at the 0.05 level of significance in G4 from 32 p.p.t. control. Different letters indicate statistical differences at P<0.05 in G8 across treatments.

View larger version (12K): [in a new window]   Fig. 9. Arginine kinase mRNA expression (relative amounts) in anterior (G4, black bars) and posterior (G8, gray bars) gills of green crabs acclimated to 32 p.p.t. and transferred to 10 p.p.t. salinity for either 4 (10-4days) or 7 days (11-7days). Values are means ± s.e.m. (N=5–6). T=12°C.