Abstract
Cells from connecting tubule and cortical collecting duct of rabbit kidney were isolated by immunodissection with mAb R2G9 and cultured on permeable filters. Confluent monolayers developed an amiloride-sensitive transepithelial potential difference of −50±1 mV (lumen negative) and a transepithelial resistance of 507±18 Ω cm2. Transepithelial Ca2+ transport increased dose-dependently with apical [Ca2+] and, in solutions containing 1 mM Ca2+, the active transcellular Ca2+ transport rate was 92±2 nmol h−1 cm−2. Transcellular Ca2+ transport was dependent on basolateral Na+ (Na +b ). Isoosmotic substitution of Na +b for N-methylglucamine resulted in a concentration-dependent decrease in Ca2+ absorption, with maximal inhibition of 67±5%. A Hill plot of the Na+-dependence yielded a coefficient of 1.9±0.4, indicating more than one Na+ site on a Na+-dependent Ca2+ transport system. In addition, the absence of Ca 2+b resulted in a significant increase in Ca2+ transport both in the presence and absence of Na +b . Added basolaterally, ouabain (0.1 mM) inhibited Ca2+ transport to the same extent as did Na+-free solutions, while bepridil (0.1 mM), an inhibitor of Na+/Ca2+ exchange, reduced Ca2+ transport by 32±6%. Methoxyverapamil, felodipine, flunarizine and diltiazem (10 μM) were without effect. Depolarisation of the basolateral membrane, by raising [K+]b to 60 mM, significantly decreased transcellular Ca2+ transport, which is indicative of electrogenic Na+/Ca2+ exchange. In conclusion, active Ca2+ transport in the collecting system of rabbit kidney is largely driven by basolateral Na+/Ca2+ exchange. However, a residual Ca2+ absorption of about 30% was always observed, suggesting that other Ca2+ transport mechanisms, presumably a Ca2+-ATPase, participate as well.
Similar content being viewed by others
References
Bacskai BJ, Friedman PA (1990) Activation of latent Ca2+ channels in renal epithelial cells by parathyroid hormone. Nature 347:388–391
Bindels RJM, Hartog A, Timmermans JAH, van Os CH (1991) Active Ca2+ transport in primary cultures of rabbit kidney collecting duct system: stimulation by 1,25-dihydroxyvitamin D3 and PTH. Am J Physiol 261:F799-F807
Borke JL, Minami J, Verma A, Penniston JT, Kumar R (1987) Monoclonal antibodies to human erythrocyte membrane Ca2+-Mg2+ adenosine triphosphate pump recognize an epitope in the basolateral membrane of human kidney distal tubule cells. J Clin Invest 80:1225–1231
Bourdeau JE, Burg MB (1980) Effect of PTH on calcium transport across the cortical thick ascending limb of Henle's loop. Am J Physiol 239:F121-F126
Bourdeau JE, Lau K (1990) Basolateral cell membrane Ca-Na exchange in single rabbit connecting tubules. Am J Physiol 258:F1497-F1503
Bourdeau JE, Eby BK (1990) cAMP-stimulated rise of [Ca2+] in rabbit connecting tubules — role of peritubular Ca. Am J Physiol 258:F751-F755
Cervetto L, Lagnado L, Perry RJ, Robinson DW, McNaughton PA (1989) Extrusion of calcium from rod outer segments is driven by both sodium and potassium gradients. Nature 337:740–743
Doucet A, Katz AI (1982) High-affinity Ca-Mg-ATPase along the rabbit nephron. Am J Physiol 242:F346-F352
Favus M (1985) Factors that influence absorption and secretion of calcium in the small intestine and colon. Am J Physiol 248:G147-G157
Flik G, Schoenmakers TJM, Groot JA, van Os CH, Wendelaar Bonga SE (1990) Calcium absorption by fish intestine: the involvement of ATP- and sodium-dependent calcium extrusion mechanisms. J Membr Biol 113:13–22
Gmaj P, Murer H, Kinne R (1979) Calcium ion transport across plasma membranes isolated from rat kidney. Biochem J 178:549–557
Kaczorowski GJ, Slaughter RS, King VF, Garcia ML (1989) Inhibitors of sodium calcium exchange. Identification and development of probes of transport activity. Biochim Biophys Acta 988:287–302
Lagnado L, McNaughton PA (1990) Electrogenic properties of the Na-Ca exchange. J Membr Biol 113:177–191
Nellans HN, Goldsmith RS (1981) Transepithelial calcium transport by rat cecum: high-efficiency absorptive site. Am J Physiol 244:G424-G431
Nicoll DA, Longoni S, Philipson KD (1990) Molecular cloning and functional expression of the cardiac sarcolemmal Na+-Ca2+ exchanger. Science 250:562–565
Nicoll DA, Barrios BR, Philipson KD (1991) Na+-Ca2+ exchangers from rod outer segments and cardiac sarcolemma: comparison of properties. Am J Physiol 260:C1212-C1216
Philipson KD (1985) Sodium-calcium exchange in plasma membrane vesicles. Annu Rev Physiol 47:561–571
Ramachandran C, Brunette MG (1989) The renal Na+-Ca2+ exchange system is located exclusively in the distal tubule. Biochem J 257:259–264
Schnetkamp PPM, Basu DK, Szerencsei RT (1989) Na+-Ca2+ exchange in bovine rod outer segments requires and transports K+. Am J Physiol 257:C153-C157
Shimizu T, Yoshitomi K, Nakamura M, Imai M (1990) Effects of PTH, calcitonin and cAMP on calcium transport in rabbit distal nephron segments. Am J Physiol 259:F408-F414
Shimizu T, Nakamura M, Yoshitomi K, Imai M (1991) Interaction of trichlormethiazide or amiloride with PTH in stimulating Ca2+ absorption in rabbit CNT. J Membr Biol 261:F36-F43
Smith PR, Benos DJ (1991) Epithelial Na+ channels. Annu Rev Physiol 53:509–530
Snedecor GW, Cochran WG (1974) Statistical methods. Iowa State University Press, Ames, Iowa
Snowdone KW, Borle AB (1985) Effects of low extracellular sodium on cytosolic ionized calcium. J Biol Chem 260:14998–15007
Spedding M (1985) Calcium antagonist subgroups. Trends Physiol Sci 6:109–114
Taniguchi S, Marchetti J, Morel F (1989) Na/Ca exchangers in collecting cells of rat kidney — a single tubule Fura-2 study. Pflügers Arch 415:191–197
Van Heeswijk MPE, Geertsen JAM, van Os CH (1984) Kinetic properties of the ATP-dependent Ca2+ pump and the Na/Ca exchange system in basolateral membranes from rat kidney cortex. J Membr Biol 79:19–31
Van Os CH (1987) Transcellular calcium transport in intestinal and renal epithelial cells. Biochim Biophys Acta 906:195–222
Wiener H, de Jong MD, van Os CH (1990) Active Ca2+ transport systems in basolateral membranes from rabbit distal colon. J Intern Med 228, S 732:119–124
Younes A, Fontanavra C, Schneider JM (1981) Effects of bepridil on the Ca2+ dependent ATPase activity of sarcoplasmic reticulum. Biochem Pharmacol 30:2979–2981
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bindels, R.J.M., Ramakers, P.L.M., Dempster, J.A. et al. Role of Na+/Ca2+ exchange in transcellular Ca2+ transport across primary cultures of rabbit kidney collecting system. Pflugers Arch. 420, 566–572 (1992). https://doi.org/10.1007/BF00374634
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00374634