Short- and long-term, salinity-induced modulation of V-ATPase activity in the posterior gills of the true freshwater crab, Dilocarcinus pagei (Brachyura, Trichodactylidae)

To better understand the biochemical mechanisms underlying anisosmotic extracellular regulation in the freshwater Brachyura, we kinetically characterized the V-ATPase from the posterior gills of Dilocarcinus pagei, acclimated for 10days to salinities up to 21‰. Specific activity was highest in fresh water (26.5±2.1U mg(-1)), decreasing in 5‰ to 21‰, attaining 3-fold less at 15‰. Apparent affinities for ATP and Mg(2+) respectively increased 3.2- and 2-fold at 10‰, suggesting expression of different isoenzymes. In a 240-h time-course study of exposure to 21‰, maximum specific activity decreased 2.5- to 4-fold within 1 to 24h while apparent affinities for ATP and Mg(2+) respectively increased by 12-fold within 24h and 2.4-fold after 1h, unchanged thereafter. K(I) for bafilomycin A(1) decreased 150-fold after 1h, remaining constant up to 120h. This is the first kinetic analysis of V-ATPase specific activity in crustacean gills during salinity acclimation. Our findings indicate active gill Cl(-) uptake by D. pagei in fresh water, and short- and long-term down-regulation of V-ATPase-driven ion uptake processes during salinity exposure, aiding in comprehension of the biochemical adaptations underpinning the establishment of the Brachyura in fresh water.