Hypoxia attenuate ionic transport in the isolated gill epithelium of Carcinus maenas
The gills are osmorespiratory organs of aquatic organisms and the prime target of environmentally induced hypoxia. We have studied the impact of severe hypoxia (0.5 mg O2/L) on the ionic transport across posterior gills of Carcinus maenas acclimated to 12 ppt seawater (DSW). The short-circuit current (Isc) across hemilamellae from gills i.e. active ion transport was studied in micro Ussing chambers. Hypoxia induced by deoxygenation of the basolateral side, and not the apical side, resulted in time-dependent inhibition of Isc and full recovery of Isc after reoxygenation. Exposure of the crabs to severe 7 h hypoxia decreased the specific activity of Na+,K+-ATPase in the gills by 36%. Full recovery of enzyme activity occurred in fasted crabs after 3 days of reoxygenation. The intensity of Western blotting bands was not different in the gills of oxygenated, hypoxic and reoxygenated crabs. The reversible, nonspecific blocker of K+ channels Cs and hypoxia inhibited over 90% of Isc which is after reoxygenation fully recovered. The specific blocker of Cl− channels NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] blocked Isc by 68.5%. Only the rest of not inhibited Isc in aerated saline was blocked by hypoxia and recovered after reoxygenation. The activity of the antioxidant enzyme catalase was not changed during hypoxia and reoxygenation kept the high enzyme activity in the gills at the level of crabs acclimated to DSW. As a response to hypoxia the presence of 2 mM H2O2 induce an initial slight transient decrease of Isc followed by a rise and after reoxygenation fully recovered Isc. Incubation of hemilamellae with the antioxidant derivative Trolox did not affect the inhibition of Isc by hypoxia.