Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1990 Feb;421:13–32. doi: 10.1113/jphysiol.1990.sp017931

Effect of Na+ and K+ on Cl- distribution in guinea-pig vas deferens smooth muscle: evidence for Na+, K+, Cl- co-transport.

C C Aickin 1, A F Brading 1
PMCID: PMC1190071  PMID: 1693397

Abstract

1. Smooth muscle cells of the guinea-pig vas deferens after Cl- depletion actively reaccumulate ions to a level many times higher than that predicted by a passive distribution, even when anion exchange (largely responsible for Cl- movements in this preparation) is inhibited by DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid). The cells therefore must possess a second mechanism for Cl- accumulation. We have now investigated the ionic requirement of this mechanism using a combination of ion analysis, 36Cl fluxes and direct measurement of the intracellular Cl- activity (aiCl). 2. In the steady state, the Cl- content of tissues was 12-16% less in Na(+)-free solution than in normal Krebs solution. 3. Loss of 36Cl into Cl(-)-free solution was slowed by the absence of Na+ and accelerated on its readdition. Uptake of 36Cl by Cl(-)-depleted tissues was also reduced in the absence of extracellular Na+, particularly at longer time intervals as uptake reached completion. These effects occurred in the presence and absence of CO2-HCO3- and in the presence of DIDS. 4. The initial rate of rise of aiCl on readdition of Cl- to Cl(-)-depleted cells was not significantly affected by the absence of Na+ in the presence of a functional anion exchange, but aiCl stabilized at a lower value than in normal solution. Readdition of Na+ stimulated a rise in aiCl to the control level. Removal and readdition of K+ under these conditions had negligible effects. 5. When anion exchange was inhibited by the presence of DIDS, removal and readdition of Na+ caused, respectively, a marked inhibition and stimulation of the rise in aiCl during Cl- reaccumulation. Under these conditions removal and readdition of K+ had similar effects. 6. The results suggest that Na+, K+, Cl- co-transport is involved in transmembrane movements of Cl- at least when the anion exchange mechanism is blocked. 7. The possibility that the marked effects of changes in external Na+ on transmembrane Cl- movements in the presence of a functional anion exchange mechanism are caused by secondary effects due to changes in intracellular pH as well as to suppression of Na+, K+, Cl- co-transport is discussed.

Full text

PDF
13

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aalkjaer C., Cragoe E. J., Jr Intracellular pH regulation in resting and contracting segments of rat mesenteric resistance vessels. J Physiol. 1988 Aug;402:391–410. doi: 10.1113/jphysiol.1988.sp017211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aickin C. C., Betz W. J., Harris G. L. Intracellular chloride and the mechanism for its accumulation in rat lumbrical muscle. J Physiol. 1989 Apr;411:437–455. doi: 10.1113/jphysiol.1989.sp017582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aickin C. C., Brading A. F. Advances in the understanding of transmembrane ionic gradients and permeabilities in smooth muscle obtained by using ion-selective micro-electrodes. Experientia. 1985 Jul 15;41(7):879–887. doi: 10.1007/BF01970005. [DOI] [PubMed] [Google Scholar]
  4. Aickin C. C., Brading A. F. Measurement of intracellular chloride in guinea-pig vas deferens by ion analysis, 36chloride efflux and micro-electrodes. J Physiol. 1982 May;326:139–154. doi: 10.1113/jphysiol.1982.sp014182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Aickin C. C., Brading A. F. The effect of loop diuretics on Cl- transport in smooth muscle of the guinea-pig vas deferens and taenia from the caecum. J Physiol. 1990 Feb;421:33–53. doi: 10.1113/jphysiol.1990.sp017932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Aickin C. C., Brading A. F. The role of chloride-bicarbonate exchange in the regulation of intracellular chloride in guinea-pig vas deferens. J Physiol. 1984 Apr;349:587–606. doi: 10.1113/jphysiol.1984.sp015175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Aickin C. C., Brading A. F. Towards an estimate of chloride permeability in the smooth muscle of guinea-pig vas deferens. J Physiol. 1983 Mar;336:179–197. doi: 10.1113/jphysiol.1983.sp014575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Aickin C. C., Brading A. F., Walmsley D. An investigation of sodium-calcium exchange in the smooth muscle of guinea-pig ureter. J Physiol. 1987 Oct;391:325–346. doi: 10.1113/jphysiol.1987.sp016741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Aickin C. C. Investigation of factors affecting the intracellular sodium activity in the smooth muscle of guinea-pig ureter. J Physiol. 1987 Apr;385:483–505. doi: 10.1113/jphysiol.1987.sp016503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Aickin C. C. Movement of acid equivalents across the mammalian smooth muscle cell membrane. Ciba Found Symp. 1988;139:3–22. doi: 10.1002/9780470513699.ch2. [DOI] [PubMed] [Google Scholar]
  11. Aickin C. C., Vermuë N. A. Microelectrode measurement of intracellular chloride activity in smooth muscle cells of guinea-pig ureter. Pflugers Arch. 1983 Apr;397(1):25–28. doi: 10.1007/BF00585163. [DOI] [PubMed] [Google Scholar]
  12. Altamirano A. A., Russell J. M. Coupled Na/K/Cl efflux. "Reverse" unidirectional fluxes in squid giant axons. J Gen Physiol. 1987 May;89(5):669–686. doi: 10.1085/jgp.89.5.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ballanyi K., Grafe P. An intracellular analysis of gamma-aminobutyric-acid-associated ion movements in rat sympathetic neurones. J Physiol. 1985 Aug;365:41–58. doi: 10.1113/jphysiol.1985.sp015758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Brading A. F. Calcium-induced increase in membrane permeability in the guinea-pig taenia coli: evidence for involvement of a sodium-calcium exchange mechanism. J Physiol. 1978 Feb;275:65–84. doi: 10.1113/jphysiol.1978.sp012178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Casteels R. The action of ouabain on the smooth muscle cells of the guinea-pig's taenia coli. J Physiol. 1966 May;184(1):131–142. doi: 10.1113/jphysiol.1966.sp007907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Casteels R. The distribution of chloride ions in the smooth muscle cells of the guinea-pig's taenia coli. J Physiol. 1971 Apr;214(2):225–243. doi: 10.1113/jphysiol.1971.sp009429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Chipperfield A. R. The (Na+-K+-Cl-) co-transport system. Clin Sci (Lond) 1986 Nov;71(5):465–476. doi: 10.1042/cs0710465. [DOI] [PubMed] [Google Scholar]
  18. Ellory J. C., Dunham P. B., Logue P. J., Stewart G. W. Anion-dependent cation transport in erythrocytes. Philos Trans R Soc Lond B Biol Sci. 1982 Dec 1;299(1097):483–495. doi: 10.1098/rstb.1982.0146. [DOI] [PubMed] [Google Scholar]
  19. Harris G. L., Betz W. J. Evidence for active chloride accumulation in normal and denervated rat lumbrical muscle. J Gen Physiol. 1987 Jul;90(1):127–144. doi: 10.1085/jgp.90.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ito Y., Kuriyama H., Parker I. Calcium transients evoked by electrical stimulation of smooth muscle from guinea-pig ileum recorded by the use of Fura-2. J Physiol. 1988 Dec;407:117–134. doi: 10.1113/jphysiol.1988.sp017406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Korbmacher C., Helbig H., Stahl F., Wiederholt M. Evidence for Na/H exchange and Cl/HCO3 exchange in A10 vascular smooth muscle cells. Pflugers Arch. 1988 Jul;412(1-2):29–36. doi: 10.1007/BF00583728. [DOI] [PubMed] [Google Scholar]
  22. Kreye V. A., Bauer P. K., Villhauer I. Evidence for furosemide-sensitive active chloride transport in vascular smooth muscle. Eur J Pharmacol. 1981 Jul 17;73(1):91–95. doi: 10.1016/0014-2999(81)90150-3. [DOI] [PubMed] [Google Scholar]
  23. Little P. J., Cragoe E. J., Jr, Bobik A. Na-H exchange is a major pathway for Na influx in rat vascular smooth muscle. Am J Physiol. 1986 Nov;251(5 Pt 1):C707–C712. doi: 10.1152/ajpcell.1986.251.5.C707. [DOI] [PubMed] [Google Scholar]
  24. Liu S., Jacob R., Piwnica-Worms D., Lieberman M. (Na + K + 2Cl) cotransport in cultured embryonic chick heart cells. Am J Physiol. 1987 Nov;253(5 Pt 1):C721–C730. doi: 10.1152/ajpcell.1987.253.5.C721. [DOI] [PubMed] [Google Scholar]
  25. O'Grady S. M., Palfrey H. C., Field M. Characteristics and functions of Na-K-Cl cotransport in epithelial tissues. Am J Physiol. 1987 Aug;253(2 Pt 1):C177–C192. doi: 10.1152/ajpcell.1987.253.2.C177. [DOI] [PubMed] [Google Scholar]
  26. Owen N. E. Regulation of Na/K/Cl cotransport in vascular smooth muscle cells. Biochem Biophys Res Commun. 1984 Dec 14;125(2):500–508. doi: 10.1016/0006-291x(84)90568-0. [DOI] [PubMed] [Google Scholar]
  27. Ozawa T., Saito Y., Nishiyama A. Mechanism of uphill chloride transport of the mouse lacrimal acinar cells: studies with Cl- -sensitive microelectrode. Pflugers Arch. 1988 Oct;412(5):509–515. doi: 10.1007/BF00582540. [DOI] [PubMed] [Google Scholar]
  28. Palfrey H. C., Rao M. C. Na/K/Cl co-transport and its regulation. J Exp Biol. 1983 Sep;106:43–54. doi: 10.1242/jeb.106.1.43. [DOI] [PubMed] [Google Scholar]
  29. Prosser C. L. Smooth muscle. Annu Rev Physiol. 1974;36:503–535. doi: 10.1146/annurev.ph.36.030174.002443. [DOI] [PubMed] [Google Scholar]
  30. Russell J. M. Cation-coupled chloride influx in squid axon. Role of potassium and stoichiometry of the transport process. J Gen Physiol. 1983 Jun;81(6):909–925. doi: 10.1085/jgp.81.6.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thomas R. C., Cohen C. J. A liquid ion-exchanger alternative to KCl for filling intracellular reference microelectrodes. Pflugers Arch. 1981 Apr;390(1):96–98. doi: 10.1007/BF00582719. [DOI] [PubMed] [Google Scholar]
  32. Turner R. J., George J. N. Cl(-)-HCO3- exchange is present with Na+-K+-Cl- cotransport in rabbit parotid acinar basolateral membranes. Am J Physiol. 1988 Mar;254(3 Pt 1):C391–C396. doi: 10.1152/ajpcell.1988.254.3.C391. [DOI] [PubMed] [Google Scholar]
  33. Vaughan-Jones R. D. An investigation of chloride-bicarbonate exchange in the sheep cardiac Purkinje fibre. J Physiol. 1986 Oct;379:377–406. doi: 10.1113/jphysiol.1986.sp016259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vaughan-Jones R. D. Chloride activity and its control in skeletal and cardiac muscle. Philos Trans R Soc Lond B Biol Sci. 1982 Dec 1;299(1097):537–548. doi: 10.1098/rstb.1982.0150. [DOI] [PubMed] [Google Scholar]
  35. Vaughan-Jones R. D. Regulation of chloride in quiescent sheep-heart Purkinje fibres studied using intracellular chloride and pH-sensitive micro-electrodes. J Physiol. 1979 Oct;295:111–137. doi: 10.1113/jphysiol.1979.sp012957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Villamil M. F., Ponce J., Amorena C., Müller A. Effect of furosemide on the ionic composition of the arterial wall. TIT J Life Sci. 1979;9(1-2):9–14. [PubMed] [Google Scholar]
  37. Weissberg P. L., Little P. J., Cragoe E. J., Jr, Bobik A. Na-H antiport in cultured rat aortic smooth muscle: its role in cytoplasmic pH regulation. Am J Physiol. 1987 Aug;253(2 Pt 1):C193–C198. doi: 10.1152/ajpcell.1987.253.2.C193. [DOI] [PubMed] [Google Scholar]
  38. Widdicombe J. H., Brading A. F. A possible role of linked Na and Cl movement in active Cl uptake in smooth muscle. Pflugers Arch. 1980 Jul;386(1):35–37. doi: 10.1007/BF00584184. [DOI] [PubMed] [Google Scholar]
  39. Wiley J. S., Cooper R. A. A furosemide-sensitive cotransport of sodium plus potassium in the human red cell. J Clin Invest. 1974 Mar;53(3):745–755. doi: 10.1172/JCI107613. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES