Ultracytochemical location of Na(+)/K(+)-atpase activity and effect of high salinity acclimation in gill and renal epithelia of the freshwater shrimp Macrobrachium olfersii (Crustacea, Decapoda).

Abstract

Accumulation sites of lead phosphate reaction product consequent to Na(+)/K(+)-ATPase activity in gill and renal epithelia of the freshwater shrimp Macrobrachium olfersii were located ultracytochemically by para-nitrophenyl-phosphate hydrolysis and lead precipitation, and quantified per unit membrane area and cytoplasmic volume. In shrimps in freshwater (<0.5 per thousand S, 20 mOsm/kg H(2)O, 0.7 mEq Na(+)/liter), numerous sites of electron-dense, Na(+)/K(+)-ATPase reaction product accumulation were demonstrated in the membrane invaginations of the mitochondria-rich, intralamellar septal cells (12.5 +/- 1.7 sites/microm(2) membrane, 179 +/- 22 sites/microm(3) cytoplasm, mean+/- SEM, N </= 7) and in the basal region of the medial renal tubules (19.8 +/- 1.8 sites/microm(2) membrane, 437 +/- 53 sites/microm(3) cytoplasm), but not in the pillar cells whose apical flanges form the primary interface with the external medium. A putative, ouabain-insensitive Na(+)- or H(+)-ATPase was found in the apical microvilli of the medial renal tubules (17.4 +/- 1.7 sites/microm(2) membrane, 629 +/- 101 sites/microm(3) cytoplasm). This restricted location of Na(+)/K(+)-ATPase activity within the gill epithelium suggests that during uptake, Na(+) moves across the apical pillar cell membrane, passes through specialized, basolateral coupling junctions into the septal cell cytoplasm and is pumped into the hemolymph via the Na(+)/K(+)-ATPase in the invagination membranes. In shrimps acclimated to a high-salinity medium (21 per thousand S, 630 mOsm/kg H(2)O, 280 mEq Na(+)/liter) for 2 and 5 days, the mean number of sites of para-nitrophenylphosphatase activity/microm(2) membrane and /microm(3) cytoplasm for both epithelia increases markedly by 83 and 163%, respectively. However, after 10 days acclimation, the number of sites declines dramatically, attaining values far below those for shrimps in freshwater. These acclimation-induced alterations in numerical density/microm(3) cytoplasm cannot be accounted for by corresponding changes in membrane surface density (microm(2) membrane/microm(3) cytoplasm) and reflect a real alteration in the number of Na(+)/K(+)-ATPase reaction product sites/unit membrane area. These data suggest that neither the gill nor the renal Na(+)/K(+)-ATPase systems function at maximal activity in shrimps in freshwater, possibly due to the low Na(+) concentration, and are initially stimulated by the increase in external ionic concentration. However, these powerful Na(+) transport systems respond to salt loading by a notable reduction in the number of hydrolysis sites, possibly through the incorporation of the Na(+)/K(+)-ATPase into isolated membrane vesicles in the basal invaginations of the medial renal tubules, together with ultrastructural alterations like the spatial isolation of the mitochondria by multiple membrane stacks in the intralamellar septal cells. J. Exp. Zool. 284:617-628, 1999.