Summary
Measurements of unidirectional calcium fluxes in stripped intestinal epithelium of the tilapia,Oreochromis mossambicus, in the presence of ouabain or in the absence of sodium indicated that calcium absorption via the fish intestine is sodium dependent. Active Ca2+ transport mechanisms in the enterocyte plasma membrane were analyzed. The maximum capacity of the ATP-dependent Ca2+ pump (V m :0.63 nmol·min−1 mg−1,K m : 27nm Ca2+) is calculated to be 2.17 nmol·min−1·mg−1, correcting for 29% inside-out oriented vesicles in the membrane preparation. The maximum capacity of the Na+/Ca2+ exchanger with high affinity for Ca2+ (V m :7.2 nmol·min−1·mg−1,K m : 181nm Ca2+) is calculated to be 13.6 nmol·min−1·mg−1, correcting for 53% resealed vesicles and assuming symmetrical behavior of the Na+/Ca2+ exchanger. The high affinity for Ca2+ and the sixfold higher capacity of the exchanger compared to the ATPase suggest strongly that the Na+/Ca2+ exchanger will contribute substantially to Ca2+ extrusion in the fish enterocyte. Further evidence for an important contribution of Na+/Ca2+ exchange to Ca2+ extrusion was obtained from studies in which the simultaneous operation of ATP-and Na+-gradient-driven Ca2+ pumps in inside-out vesicles was evaluated. The fish enterocyte appears to present a model for a Ca2+ transporting cell, in which Na+/Ca2+ exchange activity with high affinity for Ca2+ extrudes Ca2+ from the cell.
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Albus, H., Groot, J.A., Siegenbeek van Heukelom, J. 1979. Effects of glucose and ouabain on transepithelial electrical resistance and cell volume in stripped and unstripped goldfish intestine.Pfluegers Arch. 383:55–66
Bakker, R., Groot, J.A. 1984. cAMP-mediated effects of ouabain and theophylline on paracellular ion selectivity.Am. J. Physiol. 246:G213-G217
Bayerdörffer, E., Haase, W., Schulz, I. 1985. Na+/Ca2+ countertransport in plasma membrane of rat pancreatic acinar cells.J. Membrane Biol. 87:107–119
Berg, A. 1970. Studies on the metabolism of calcium and strontium in freshwater fish. II. Relative contribution of direct and intestinal absorption in growth conditions.Mem. Ist. Ital. Idrobiol. Dott Marco de Marchi 26:241–255
Cheon, J., Reeves, J.P. 1988. Sodium-calcium exchange in membrane vesicles fromArtemia.Arch. Biochem. Biophys. 267:736–741
Cook, N.J., Kaupp, U.B. 1988. Solubilization, purification, and reconstitution of the sodium-calcium exchanger from bovine retinal rod outer segments.J. Biol. Chem. 263:11382–11388
Corven, E.J.J.M., Roche, C., Van, Os, C.H. 1985. Distribution of Ca2+-ATPase, ATP-dependent Ca2+ transport, calmodulin and vitamin D-dependent Ca2+ binding protein along the villus-crypt axis in rat duodenum.Biochim. Biophys. Acta 820:274–282
Eisenmann, G., Sandblom, J., Neher, E. 1978. Interactions in cation permeation through the gramicidin channel. Cs, Rb, K, Na, Li, Tl, H and effects of anion binding.Biophys. J. 22:307–340
Favus, M.J., Angeid-Backman, E., Breyer, M.D., Coe, F.L. 1983. Effects of trifluoperazine, ouabain and ethacrynic acid on intestinal calcium transport.Am. J. Physiol. 244:G111-G115
Flik, G., Fenwick, J.C., Kolar, Z., Mayer-Gostan, N., Wendelaar Bonga, S.E. 1985a. Whole-body calcium flux rates in cichlid teleost fishOreochromis mossambicus adapted to freshwater.Am. J. Physiol. 249:R432-R437
Flik, G., Fenwick, J.C., Wendelaar Bonga, S.E. 1989. Calcitropic actions of prolactin in freshwater North American eel (Auguilla rostrata LeSueur).Am. J. Physiol. 257:R74-R79
Flik, G., Perry, S.F. 1989. Cortisol stimulates whole body calcium uptake and the branchial calcium pump in freshwater rainbow trout.J. Endocrinol. 120:75–82
Flik, G., Van Rijs, J.H., Wendelaar Bonga, S.E. 1985b. Evidence for high-affinity Ca2+-ATPase activity and ATP-driven Ca2+-transport in membrane preparations of the gill epithelium of the cichlid fishOreochromis mossambicus.J. Exp. Biol. 119:335–347
Flik, G., Wenelaar Bonga, S.E., Fenwick, J.C. 1985c. Active Ca2+ transport in plasma membranes of branchial epithelium of the North-American eel,Anguilla rostrata LeSueur.Biol. Cell. 55:265–272
Garcia, M.L., Slaughter, R.S., King, V.F., Kaczorowski, G.J. 1988. Inhibition of sodium-calcium exchange in cardiac sarcolemmal membrane vesicles. 2. Mechanism of inhibition by bepridil.Biochemistry 27:2410–2415
Ghijsen, W.E.J.M., De Jong, M.D., Van Os, C.H. 1983. Kinetic properties of Na+/Ca2+ exchange in basolateral plasma membranes of rat small intestine.Biochim. Biophys. Acta 730:85–94
Gill, D.L., Chueh, S-H., Whitlow, C.L. 1984. Functional importance of the synaptic plasma membrane calcium pump and sodium-calcium exchanger.J. Biol. Chem. 259:10807–10813
Groot, J.A., Albus, H., Bakker, R. 1981. Analysis of the ouabain induced increase in transepithelial electrical resistance in the goldfish intestinal mucosa.Pfluegers Arch. 392:67–71
Heeswijk, M.P.M. van, Geertsen, J.A.M., Os, C.H. van 1984. Kinetic properties of the ATP-dependent Ca2+ pump and the Na+/Ca2+ exchange system in basolateral membranes from rat kidney cortex.J. Membrane Biol. 79:19–31
Hildmann, B., Schmidt, A., Murer, H. 1982. Ca2+-transport across basal-lateral plasma membranes from rat small intestinal epithelial cells.J. Membrane Biol. 65:55–62
Jayakumar, A., Cheng, L., Liang, C.T., Sacktor, B. 1984. Sodium gradient-dependent calcium uptake in renal basolateral membrane vesicles.J. Biol. Chem. 259:10827–10833
Leatherbarrow, R.J. 1987. A non-linear regression analysis program for the IBM PC. Elsevier Biosoft, Amsterdam
Moore, L., Fitzpatrick, D.F., Chen, T.S., Landon, E.J. 1974. Calcium pump activity of the renal plasma membrane and renal microsomes.Biochim. Biophys. Acta. 345:405–418
Murer, H., Hildmann, B. 1981. Transcellular transport of calcium and inorganic phosphate in the small intestinal epithelium.Am. J. Physiol. 240:G409-G416
Ortiz, O.E., Sjodin, R.A. 1984. Sodium- and adenosine-triphosphate-dependent calcium movements in membrane vesicles prepared from dog erythrocytes.J. Physiol. (London) 354:287–301
Perry, S.F., Flik, G. 1988. Characterization of branchial transepithelial calcium fluxes in freshwater trout,Salmo gairdneri.Am. J. Physiol. 254:R491-R498
Philipson, K.D. 1985. Symmetry properties of the Na+−Ca2+ exchange mechanism in cardiac sarcolemmal vesicles.Biochim. Biophys. Acta 821:367–376
Philipson, K.D., Nishimoto, A.Y. 1982. Na+−Ca2+ exchange in inside-out cardiac sarcolemmal vesicles.J. Biol. Chem. 257:5111–5117
Philipson, K.D., Ward, R. 1986. Ca2+ transport capacity of sarcolemmal Na+−Ca2+ exchange. Extrapolation of vesicle data to in vivo conditions.J. Mol. Cell. Cardiol. 18:943–951
Reeves, J. P., Hale, C. C. 1984. The stoichiometry of the cardiac sodium-calcium exchange system.J. Biol. Chem. 259:7733–7739
Reeves, J.P., Sutko, J.L. 1983. Competitive interactions of sodium and calcium with the sodium-calcium exchange system of cardiac sarcolemmal vesicles.J. Biol. Chem. 258:3178–3182
Sandblom, J., Eisenman, G., Neher, E. 1977. Ionic selectivity, saturation and block in gramicidin A channels: I. Theory for the electrical properties of ion selective channels having two pairs of binding sites and multiple conductance states.J. Membrane Biol. 31:383–417
Sillen, L.G., Martell, A.E. 1964. Stability constants of metal ion complexes. The Chemical Society, Special Publication no. 17, London
Slaughter, R.S., Garcia, M.L., Cragoe, E.J., Jr., Reeves, J.P., Kaczorowski, G.J. 1988. Inhibition of sodium-calcium exchange in cardiac sarcolemmal membrane vesicles. I. Mechanism of inhibition by amiloride analogues.Biochemistry 27:2403–2409
Spedding, M. 1985. Activators and inactivators of Ca2+ channels: New perspectives.J. Pharmacol. (Paris) 16:319–343
Van Os, C.H. 1987. Transcellular calcium transport in intestinal and renal epithelial cells.Biochim. Biophys. Acta 906:195–222
Verbost P.M., Flik, G., Lock, R.A.C., Bonga, S.E.W. 1988. Cadmium inhibits plasma membrane calcium transport.J. Membrane Biol. 102:97–104
Verbost, P.M., Flik, G., Pang, P.K.T., Lock, R.A.C., Wendelaar Bonga, S.E. 1989. Cadmium inhibition of the erythrocyte Ca2+ pump. A molecular interpretation.J. Biol. Chem. 264:5613–5615
Younes, A., Fontanarava, C., Schneider, J.M. 1981. Effects of bepridil on the Ca2+ dependent ATPase activity of sarcoplasmic reticulum.Biochem. Pharmacol. 30:2979–2981
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Flik, G., Schoenmakers, T.J.M., Groot, J.A. et al. Calcium absorption by fish intestine: The involvement of ATP-and sodium-dependent calcium extrusion mechanisms. J. Membrain Biol. 113, 13–22 (1990). https://doi.org/10.1007/BF01869601
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DOI: https://doi.org/10.1007/BF01869601