BASOLATERAL TRANSPORT OF TAURINE IN EPITHELIAL CELLS OF ISOLATED, PERFUSED MYTILUS CALIFORNIANUS GILLS

摘要

We found that the basolateral surface of the gill epithelium of the marine mussel Mytilus californianus possesses a carrier-mediated process capable of concentrating taurine within epithelial cells. We used retrograde perfusion of gill sections to demonstrate the kinetics, specificity and ion-dependence of taurine transport, [H-3]taurine was concentrated relative to a space marker ([C-14]mannitol); this accumulation was blocked by the inclusion of 10 mmoll(-1) unlabeled taurine in the perfusate. The drop in [3H]taurine uptake at increasing concentrations of unlabeled taurine was fitted to Michaelis-Menten kinetics and indicated a basolateral process with a taurine concentration at which transport is half-maximal (K-t) of 35.3 mu moll(-1) and a maximal flux (J(max)) of 0.35 mu mol g(-1) wet mass h(-1). Taurine accumulation on the apical surface had a higher affinity (K-t=9.5 mu moll(-1)) and a higher maximum rate of transport (J(max)=1.23 mu mol g(-1) h(-1)). Basolateral transport was inhibited by inclusion in the perfusate of 1 mmoll(-1) of another alpha-amino acid (alpha-alanine), but not by inclusion of alpha-alanine, glutamic acid or betaine. The dependence of basolateral taurine transport on Na+ (when replaced with N-methyl-D-glucamine) was sigmoidal with an apparent Hill coefficient of 2.3, indicating that more than one Na+ is necessary for the transport of each taurine molecule. Complete substitution of Cl- in bathing media reduced taurine accumulation by 90 % and 70 % on the apical and basolateral surfaces, respectively. Taurine accumulation on both surfaces was reduced by only 20 % when Cl- was reduced from 496 to 73 mmoll(-1), suggesting that taurine uptake is not significantly influenced by the changes in Cl- concentration accompanying the salinity fluctuations normally encountered by mussels. We estimate that the various Na+ and Cl- gradients naturally encountered by epithelial cells are capable of providing ample energy to maintain a high intracellular concentration of taurine. We suggest that the ability of epithelial cells to accumulate taurine across the basolateral surface from the hemolymph plays a significant role in the intracellular regulation of this important osmolyte and may effect osmolality-dependent changes in the intracellular concentration of taurine. [References: 32]