Abstract
Investigations of corneal endothelium were made to resolve the apparent contradiction of the presence of sodium/bicarbonate cotransporter (NBC) in fresh and cultured cells and NBC's reported absence in isolated plasma membrane vesicles. Gradient-driven ion fluxes into the vesicles were measured. Short-term incubations (0-30 s) showed the presence of a bicarbonate-dependent inward sodium flux (BDSF), which was active when the insides of the vesicles were preloaded with chloride ions. The BDSF was absent if chloride was present only externally to the vesicles. Chloride at concentrations between 30 and 40 mM inside the vesicle had its maximum effect on BDSF. Other anions (acetate, thiocyanate, or gluconate) inside the vesicles did not mimic the chloride effect. Associated with the net inward sodium flux was a net inward bicarbonate flux. Hill plots of sodium influx with respect to external bicarbonate concentrations indicated that the stoichiometry of the net transfer was 1.7 +/- 0.2 (mean +/- standard error, n = 5) bicarbonate ions for each sodium ion transported. There was no net chloride flux found across the membrane vesicles. The finding of a novel chloride-activated NBC activity fully resolves the apparent contradiction between whole-cell and membrane vesicle preparations.
Full Text
The Full Text of this article is available as a PDF (61.4 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aronson P. S., Suhm M. A., Nee J. Interaction of external H+ with the Na+-H+ exchanger in renal microvillus membrane vesicles. J Biol Chem. 1983 Jun 10;258(11):6767–6771. [PubMed] [Google Scholar]
- Bonanno J. A., Giasson C. Intracellular pH regulation in fresh and cultured bovine corneal endothelium. I. Na+/H+ exchange in the absence and presence of HCO3-. Invest Ophthalmol Vis Sci. 1992 Oct;33(11):3058–3067. [PubMed] [Google Scholar]
- Bonanno J. A., Giasson C. Intracellular pH regulation in fresh and cultured bovine corneal endothelium. II. Na+:HCO3- cotransport and Cl-/HCO3- exchange. Invest Ophthalmol Vis Sci. 1992 Oct;33(11):3068–3079. [PubMed] [Google Scholar]
- Boron W. F., Boulpaep E. L. Intracellular pH regulation in the renal proximal tubule of the salamander. Basolateral HCO3- transport. J Gen Physiol. 1983 Jan;81(1):53–94. doi: 10.1085/jgp.81.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eiam-Ong S., Hilden S. A., Johns C. A., Madias N. E. Stimulation of basolateral Na(+)-HCO3- cotransporter by angiotensin II in rabbit renal cortex. Am J Physiol. 1993 Aug;265(2 Pt 2):F195–F203. doi: 10.1152/ajprenal.1993.265.2.F195. [DOI] [PubMed] [Google Scholar]
- Grassl S. M., Aronson P. S. Na+/HCO3-co-transport in basolateral membrane vesicles isolated from rabbit renal cortex. J Biol Chem. 1986 Jul 5;261(19):8778–8783. [PubMed] [Google Scholar]
- Hagenbuch B., Stange G., Murer H. Sodium-bicarbonate cotransport occurs in rat kidney cortical membranes but not in rat small intestinal basolateral membranes. Biochem J. 1987 Sep 1;246(2):543–545. doi: 10.1042/bj2460543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hodson S. Evidence for a bicarbonate-dependent sodium pump in corneal endothelium. Exp Eye Res. 1971 Jan;11(1):20–29. doi: 10.1016/s0014-4835(71)80060-x. [DOI] [PubMed] [Google Scholar]
- Hodson S., Miller F. The bicarbonate ion pump in the endothelium which regulates the hydration of rabbit cornea. J Physiol. 1976 Dec;263(3):563–577. doi: 10.1113/jphysiol.1976.sp011645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huff J. W., Green K. Characteristics of bicarbonate, sodium, and chloride fluxes in the rabbit corneal endothelium. Exp Eye Res. 1983 Apr;36(4):607–615. doi: 10.1016/0014-4835(83)90054-4. [DOI] [PubMed] [Google Scholar]
- Jentsch T. J., Keller S. K., Koch M., Wiederholt M. Evidence for coupled transport of bicarbonate and sodium in cultured bovine corneal endothelial cells. J Membr Biol. 1984;81(3):189–204. doi: 10.1007/BF01868713. [DOI] [PubMed] [Google Scholar]
- Jentsch T. J., Korbmacher C., Janicke I., Fischer D. G., Stahl F., Helbig H., Hollwede H., Cragoe E. J., Jr, Keller S. K., Wiederholt M. Regulation of cytoplasmic pH of cultured bovine corneal endothelial cells in the absence and presence of bicarbonate. J Membr Biol. 1988 Jul;103(1):29–40. doi: 10.1007/BF01871930. [DOI] [PubMed] [Google Scholar]
- Jentsch T. J., Stahlknecht T. R., Hollwede H., Fischer D. G., Keller S. K., Wiederholt M. A bicarbonate-dependent process inhibitable by disulfonic stilbenes and a Na+/H+ exchange mediate 22Na+ uptake into cultured bovine corneal endothelium. J Biol Chem. 1985 Jan 25;260(2):795–801. [PubMed] [Google Scholar]
- Lane J. R., Wigham C. G., Hodson S. A. Determination of Na+/Cl-, Na+/HCO3- and Na+/K+/2Cl- co-transporter activity in corneal endothelial cell plasma membrane vesicles. Biochim Biophys Acta. 1997 Sep 4;1328(2):237–242. doi: 10.1016/s0005-2736(97)00101-6. [DOI] [PubMed] [Google Scholar]
- Lane J. R., Wigham C. G., Hodson S. A. Sodium ion uptake into isolated plasma membrane vesicles: indirect effects of other ions. Biophys J. 1999 Mar;76(3):1452–1456. doi: 10.1016/S0006-3495(99)77305-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lynch A. M., McGivan J. D. Evidence for a single common Na+-dependent transport system for alanine, glutamine, leucine and phenylalanine in brush-border membrane vesicles from bovine kidney. Biochim Biophys Acta. 1987 May 29;899(2):176–184. doi: 10.1016/0005-2736(87)90398-1. [DOI] [PubMed] [Google Scholar]
- Renner E. L., Lake J. R., Scharschmidt B. F., Zimmerli B., Meier P. J. Rat hepatocytes exhibit basolateral Na+/HCO3- cotransport. J Clin Invest. 1989 Apr;83(4):1225–1235. doi: 10.1172/JCI114005. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Riley M. V., Winkler B. S., Starnes C. A., Peters M. I. Fluid and ion transport in corneal endothelium: insensitivity to modulators of Na(+)-K(+)-2Cl- cotransport. Am J Physiol. 1997 Nov;273(5 Pt 1):C1480–C1486. doi: 10.1152/ajpcell.1997.273.5.C1480. [DOI] [PubMed] [Google Scholar]
- Sanchís D., Alemany M., Remesar X. L-alanine transport in small intestine brush-border membrane vesicles of obese rats. Biochim Biophys Acta. 1994 Jun 22;1192(2):159–166. doi: 10.1016/0005-2736(94)90114-7. [DOI] [PubMed] [Google Scholar]
- Seki G., Coppola S., Frömter E. The Na(+)-HCO3- cotransporter operates with a coupling ratio of 2 HCO3- to 1 Na+ in isolated rabbit renal proximal tubule. Pflugers Arch. 1993 Dec;425(5-6):409–416. doi: 10.1007/BF00374866. [DOI] [PubMed] [Google Scholar]
- Soleimani M., Aronson P. S. Ionic mechanism of Na+-HCO3- cotransport in rabbit renal basolateral membrane vesicles. J Biol Chem. 1989 Nov 5;264(31):18302–18308. [PubMed] [Google Scholar]
- Usui T., Seki G., Amano S., Oshika T., Miyata K., Kunimi M., Taniguchi S., Uwatoko S., Fujita T., Araie M. Functional and molecular evidence for Na(+)-HCO3- cotransporter in human corneal endothelial cells. Pflugers Arch. 1999 Sep;438(4):458–462. doi: 10.1007/s004249900081. [DOI] [PubMed] [Google Scholar]
- Wiederholt M., Jentsch T. J., Keller S. K. Electrogenic sodium-bicarbonate symport in cultured corneal endothelial cells. Pflugers Arch. 1985;405 (Suppl 1):S167–S171. doi: 10.1007/BF00581801. [DOI] [PubMed] [Google Scholar]
- Wigham C. G., Kaila D., Hodson S. A. Determination of pathways for sodium movement across corneal endothelial cell derived plasma membrane vesicles. Biochim Biophys Acta. 1994 Nov 23;1196(1):88–92. doi: 10.1016/0005-2736(94)90299-2. [DOI] [PubMed] [Google Scholar]
- Wigham C. G., Turner H. C., Ogbuehi K. C., Hodson S. A. Two pathways for electrogenic bicarbonate ion movement across the rabbit corneal endothelium. Biochim Biophys Acta. 1996 Feb 21;1279(1):104–110. doi: 10.1016/0005-2736(95)00232-4. [DOI] [PubMed] [Google Scholar]
- Wigham C., Hodson S. Bicarbonate and the trans-endothelial short circuit current of human cornea. Curr Eye Res. 1981;1(5):285–290. doi: 10.3109/02713688108999449. [DOI] [PubMed] [Google Scholar]
- Wigham C., Hodson S. The movement of sodium across short-circuited rabbit corneal endothelium. Curr Eye Res. 1985 Dec;4(12):1241–1245. doi: 10.3109/02713688509017682. [DOI] [PubMed] [Google Scholar]
- Winkler B. S., Riley M. V., Peters M. I., Williams F. J. Chloride is required for fluid transport by the rabbit corneal endothelium. Am J Physiol. 1992 May;262(5 Pt 1):C1167–C1174. doi: 10.1152/ajpcell.1992.262.5.C1167. [DOI] [PubMed] [Google Scholar]
- van Amelsvoort J. M., Sips H. J., van Dam K. Sodium-dependent alanine transport in plasma-membrane vesicles from rat liver. Biochem J. 1978 Sep 15;174(3):1083–1086. doi: 10.1042/bj1741083. [DOI] [PMC free article] [PubMed] [Google Scholar]
