Abstract
Ca2+ transport across the cortical thick ascending limb of Henle's loop (TALH) was studied on the isolated rabbit renal tubule perfused in vitro. Both the efflux (Ke) and influx coefficient (Ki) of Ca2+ were determined with45Ca at three different levels of the transtubular electrical potential difference (PDt) caused by varying the transtubular Na+ concentration gradient. The flux ratios,Ke/Ki, always exceeded those predicted from observed PDt by simple passive diffusion, and the net Ca2+ efflux was shown to occur against an electro-chemical potential. An increase in Ca2+ concentration in the perfusate was associated with a decrease inKe. Efflux of Ca2+, therefore, tended to be saturated as luminal Ca2+ concentration was increased.Ke (10−7 cm2/s) decreased from 4.13±0.56 to 2.02±0.30 (P<0.02) along with a decrease in PDt when 10−3 M NaCN was added to the bathing fluid. By contrast, 10−3 M iodoacetamide did not affectKe in spite of a significant decrease in PDt. Similarly, neither 10−5 M ouabain added to the bathing fluid nor 10−4 M ruthenium red, added either to the bathing fluid or the perfusate, affectedKe despite significant decreases in PDt. Addition of 10−5 M furosemide in the lumen caused a decrease inKe from 4.31±0.43 to 3.17±0.25 (P<0.01) in association with a decrease in PDt from 6.3±1.04 to 1.7±0.37 mV. These findings suggest; (a) Ca2+ transport across the TALH is mainly an active process requiring aerobic metabolism; (b) dissociation of Ca2+ and Cl− transport may occur under some experimental conditions.
Similar content being viewed by others
References
Burg, M. B., Green, N.: Function of the thick ascending limb of Henle's loop. Am. J. Physiol.224, 659–668 (1973)
Burg, M., Grantham, J., Abramow, M., Orloff, J.: Preparation and study of fragments of single rabbit nephrons. Am. J. Physiol.210, 1293–1298 (1966)
Carofoli, E., Sacktor, B.: The effects of ruthenium red on reactions of blowfly flight muscle mitochondria with calcium. Biochem. Biophys. Res. Commun.49, 1498–1503 (1972)
Curran, P. F., Schultz, S. G.: Transport across membranes: General principles: Handbook of physiology (Code, C., ed.), Sect. 6. The alimentary canal, Vol. III. Chapt. 65, pp. 1217–1243. Washington D.C.: American Physiological Society 1968
De Rouffignac, C., Morel, F., Moss, N., Roinel, N.: Micropuncture study of water and electrolyte movements along the loop of Henle in Psammomys with special reference to magnesium, calcium, and phosphorus. Pflügers Arch.334, 309–326 (1973)
Edwards, B. R., Baer, P. G., Sutton, R. A. L., Dirks, J. H.: Micropuncture study of diuretic effects on sodium and calcium reabsorption in the dog nephron. J. Clin. Invest.52, 2418–2427 (1973)
Edwards, B. R., Sutton, R. A. L., Dirks, J. H.: Effect of calcium infusion on renal tubular reabsorption in the dog kidney. Am. J. Physiol.227, 13–18 (1974)
Goldberg, M., Agus, Z. S., Goldfarb, S.: Renal handling of phosphate, calcium, and magnesium. In: Kidney (Brenner, B. M., and Rector, F. C. Jr., ed.), pp. 344–390. Philadelphia-London-Toronto: Saunders 1976
Grantham, J. J., Berg, M. B.: Effect of vasopressin and cyclic AMP on permeability of isolated collecting tubules. Am. J. Physiol.221, 225–259 (1966)
Ichida, S., Kuo, C. H., Matsuda, T., Yoshida, H.: Effects of La3+, Mn2+ and ruthenium red on Mg·Ca-ATPase activity and ATP-dependent Ca-binding of the synaptic plasma membrane. Jap. J. Pharmacol.26, 29–43 (1976)
Imai, M.: Function of the thin ascending limb of Henle of rats and hamsters perfused in vitro. Am. J. Physiol.232, F201-F209 (1977)
Imai, M.: Effect of bumetanide and furosemide on the thick ascending limb of Henle's loop of rabbits and rats perfused in vitro. Eur. J. Pharmacol.41, 409–416 (1977)
Imai, M., Kokko, J. P.: Sodium, chloride, urea, and water transport in the thin ascending limb of Henle. Generation of osmotic gradients by passive diffusion of solutes. J. Clin. Invest.53, 393–402 (1974)
Jamison, R. L., Frey, N. R., Lacy, F. B.: Calcium reabsorption in the thin loop of Henle. Am. J. Physiol.227, 745–751 (1974)
Lassiter, W. E., Gottschalk, C. W., Mylle, M.: Micropuncture study of renal tubular reabsorption of calcium in normal rodents. Am. J. Physiol.204, 771–775 (1963)
Martinez-Maldonado, M., Eknoyan, G., Suki, W. N.: Importance of aerobic and anaerobic metabolism in renal concentration and dilution. Am. J. Physiol.218, 1076–1081 (1970)
Moore, C. L., Korey, S. R.: Specific inhibition of mitochondrial Ca2+ transport by ruthenium red. Biochem. Biophys. Res. Commun.42, 298–305 (1971)
Moore, L., Fitzpatrick, D. F., Chen, T. S., Landon, E. J.: Calcium pump activity of the renal plasma membrane and renal microsomes. Biochim. Biophys. Acta345, 405–418 (1974)
Murayama, Y., Morel, F., LeGrimellec, C.: Phosphate, calcium and magnesium transfers in proximal tubules and loops of Henle, as measured by single nephron microperfusion experiments in the rat. Pflügers Arch.333, 1–16 (1972)
Palmer, R. F., Posey, V. A.: Calcium and adenosine triphosphate binding to renal membranes. J. Gen. Physiol.55, 89–103 (1970)
Parkinson, D. K., Radde, I. C.: Properties of Ca2+ and Mg2+-activated ATP-hydrolysing enzyme in rat kidney cortex. Biochim. Biophys. Acta242, 238–246 (1971)
Poujeol, Pl, Chabardes, D., Roinel, N., De Rouffignac, C.: Influence of extracellular fluid volume expansion on magnesium, calcium and phosphate handling along the rat nephron. Pfügers Arch.365, 203–212 (1976)
Rocha, A. S., Kokko, J. P.: Sodium chloride and water transport in the medullary thick ascending limb of Henle. Evidence for active chloride transport. J. Clin. Invest.52, 612–623 (1973)
Rocha, A. S., Magaldi, J. B., Kokko, J. P.: Calcium and phosphate transport in isolated segments of rabbit Henle's loop. J. Clin. Invest.59, 975–983 (1977)
Rossi, C. S., Vasington, F. D., Carafoli, E.: The effect of ruthenium red on the uptake and release of Ca2+ by mitochondria. Biochem. Biophys. Res. Commun.50, 846–852 (1973)
Sutton, R. A. L., Wong, N. L. M., Dirks, J. H.: Effects of parathyroid hormone on sodium and calcium transport in the dog nephron. Clin. Sci. Mol. Med.51, 345–351 (1976)
Taylor, A. N., Wasserman, R. H.: Vitamin D-induced calcium binding protein: comparative aspects in kidney and intestine. Am. J. Physiol.223, 110–114 (1972)
Ussing, H. H.: Distinction by means of tracers between active transport and diffusion. The transfer of iodide across the isolated frog skin. Acta Physiol. Scand.19, 43–56 (1950)
Vale, M. G. P., Carvalho, A. P.: Effects of ruthenium red on Ca2+ uptake and ATPase of sarcoplasmic reticulum of rabbit skeletal muscle. Biochim. Biophys. Acta325, 29–37 (1973)
Vasington, F. D., Gazzotti, P., Tiozzo, R., Carafoli, E.: The effect of ruthenium red on Ca2+ transport and respiration in rat liver mitochondria. Biochim. Biophys. Acta256, 43–54 (1972)
Watson, E. L., Vincenzi, F. F., Davis, P. W.: Ca2+-activated membrane ATPase: selective inhibition by ruthenium red. Biochim. Biophys. Acta249, 606–610 (1971)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Imai, M. Calcium transport across the rabbit thick ascending limb of Henle's loop perfused in vitro. Pflugers Arch. 374, 255–263 (1978). https://doi.org/10.1007/BF00585603
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00585603