Abstract
Rabbit cortical thick ascending limb segments were perfused in vitro, and intracellular Cl− activity was estimated in three types of experiments using conventional and chloride selective microelectrodes. In series 1 Ringer like solutions were present on the two epithelial sides. In series 2 lumen Cl− was replaced by gluconate, and in series 3 furosemide, 10–20 · 10−6 mol · 1−1, was added to the lumen perfusate. It was found that under control conditions intracellular Cl− activity, as estimated from the difference of the reading of the conventional (n=53) and ion selective electrodes (n=118) was 26±1 mmol · 1−1. This value is approximately three times higher than expected for passive distribution of Cl−1. After removal of lumen Cl− (series 2) intracellular Cl− activity fell to 9 mmol · 1−1 which is only some 4 mmol · 1−1 above passive distribution. We argue that these 4 mmol · 1−1 reflect mainly the interference with the Cl− electrode by other anions, such as phosphate. The above estimates for intracellular Cl− activity, have to be diminished by these 4 mmol·1−1, and, thus, are close to 22 mmol·1−1. In series 3 a rapid and reversible fall in intracellular Cl− from 23 to 7 mmol·1−1 was observed. We conclude that the Cl− activity in cTAL cells is clearly above equilibrium under control conditions and that it falls rapidly to values close to equilibrium when Cl− reabsorption is blocked by either removing lumen Cl− or by blocking the Cl− entry via the Na+−2Cl−−K+-carrier with furosemide. The present data directly support our concept that the furosemide induced hyperpolarization of PDb is the result of the falling cellular Cl− activity.
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References
Baumgarten CM (1981) An improved liquid ion exchanger for chloride ionselective microelectrodes. Am J Physiol 241:C258-C263
Beck F, Bauer R, Bauer U, Mason J, Dörge A, Rick R, Thurau K (1980) Electron microprobe analysis of intracellular elements in the rat kidney. Kidney Int 17:756–763
Biagi B, Sohtell M, Giebisch G (1981) Intracellular potassium activity in the rabbit proximal straight tubule. Am J Physiol 241:F677-F686
Burg MB (1982) Thick ascending limb of Henle's loop. Kidney Int 22:454–464
Burg MB, Stoner L (1976) Renal tubular chloride transport and the mode of action of some diuretics. Ann Rev Physiol 38:37–45
Burg MB, Grantham J, Abramow M, Orloff J (1966) Preparation and study of fragments of single nephrons. Am J Physiol 210(6):1293–1298
Cassola AC, Mollenhauer M, Frömter E (1982) The intracellular chloride activity of rat kidney. Pflügers Arch [submitted]
Cemerikic D, Wilcox CS, Giebish G (1982) Intracellular potential and K+ activity in rat kidney proximal tubular cells in acidosis and K+ depletion. J Membr Biol 69:159–165
Edelman A, Bouthier M, Anagnostopoulos T (1981) Chloride distribution in the proximal convoluted tubule of Necturus kidney. J Membr Biol 62:7–17
Edelman A, Curci S, Samarzija I, Frömter E (1978) Determination of intracellular K+ activity in rat kidney proximal tubular cells. Pflügers Arch 378:37–45
Frömter E, Rumrich G, Ullrich KJ (1973) Phenomenologic description of Na+, Cl− and HCO3 absorption from proximal tubules of the rat kidney. Pflügers Arch 343:189–220
Fromm M, Weskamp P, Hegel U (1980) Versatile piezoelectric driver for cell puncture. Pflügers Arch 384:69–73
Greger R (1981) Cation selectivity of the isolated perfused cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 390:30–37
Greger R (1981) Chloride reabsorption in the rabbit cortical thick ascending limb of the loop of Henle. Pflügers Arch 390:38–43
Greger R (1981) Coupled transport of Na+ and Cl− in the thick ascending limb of Henle's loop of rabbit nephron. Scand Audiol Suppl 14:1–14
Greger R, Hampel W (1981) A modified system for in vitro perfusion of isolated renal tubules. Pflügers Arch 389:175–176
Greger R, Schlatter E (1981) Presence of luminal K+, a prerequisite for active NaCl transport in the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 392:92–94
Greger R, Schlatter E (1983) Properties of the basolateral membrane of the cortical thick ascending limb of Henle's loop of rabbit kidney. A model for secondary active chloride transport. Pflügers Arch 396:325–334
Greger R, Schlatter E (1983) Properties of the lumen membrane of the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 396:315–324
Greger R, Schlatter E, Lang F (1983) Evidence for electroneutral sodium chloride cotransport in the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 396:308–314
Guggino WB, Boulpaep EL, Giebisch G (1982) Electrical properties of chloride transport across the Necturus proximal tubule. J Membr Biol 65:185–196
Hebert SC, Culpepper RM, Andreoli TE (1981) NaCl transport in mouse medullary thick ascending limb. I. Functional nephron heterogeneity and ADH-stimulated NaCl cotransport. Am J Physiol 241:F412-F431
Heinz E, Grassl S (1981) Interference of furosemide and other anion transport inhibitors with liquid Cl− exchanger electrodes. Biophysical J 33:T-PM-G11
Khuri RN, Agulian SK, Bogharian K (1974) Electrochemical potentials of chloride in distal renal tubule of the rat. Am J Physiol 227(6):1352–1355
Oberleithner H, Lang F, Wang W, Giebisch G (1982) Effects of inhibition of chloride transport on intracellular sodium activity in distal amphibian nephron. Pflügers Arch 394:55–60
Oberleithner H, Guggino W, Giebisch G (1982) Mechanism of distal tubular chloride transport in Amphiuma kidney. Am J Physiol 242:F331-F339
Schlatter E, Greger R (1982) Metabolic substrates for maintaining active NaCl transport in the isolated cortical thick ascending limb (cTAL) of rabbit kidney. Pflügers Arch 394:64
Schlatter E, Greger R, Weidtke C (1983) Effect of “high ceiling” diuretics on active salt transport in the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 396:210–217
Shindo T, Spring KR (1981) Chloride movement across the basolateral membrane of proximal tubule cells. J Membr Biol 58:35–42
Spring KR, Kimura G (1978) Chloride reabsorption by renal proximal tubules of Necturus. J Membr Biol 38:233–254
Stoner LC (1977) Isolated, perfused amphibian renal tubules: the diluting segment. Am J Physiol 233(5):F438-F444
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Greger, R., Oberleithner, H., Schlatter, E. et al. Chloride activity in cells of isolated perfused cortical thick ascending limbs of rabbit kidney. Pflugers Arch. 399, 29–34 (1983). https://doi.org/10.1007/BF00652518
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DOI: https://doi.org/10.1007/BF00652518