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Regulation of intracellular pH in cultured bovine retinal pigment epithelial cells

  • Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands
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Abstract

Regulation of intracellular pH (pHi) in bovine retinal pigment epithelium (RPE) was investigated in cell culture. pHi was measured using the pH-sensitive absorbance of intracellularly trapped 5 (and 6)-carboxy-dimethyl-fluorescein (CDMF). (1) Regulation of pHi after induction of an acid load by removal of NH4Cl could be blocked either totally by removal of extracellular sodium, or subtotally (about 90%) by application of amiloride (1 mmol/l). Additional flux measurements revealed a dose-dependent, amiloride-sensitive22Na+-uptake into Na+-loaded cells. Both results suggest the presence of a Na+/H+ antiport.

(2) When alkalinization of the cells was induced by preincubation with 50 mmol/l acetate in HCO −3 -Ringer's and subsequent removal of the weak acid, the following regulation was dependent on the presence of extracellular chloride. This process could be blocked with DIDS (1 mmol/l), suggesting the presence of a Cl−/HCO −3 exchange mechanism.

(3) We found no evidence for a Na+/HCO −3 -cotransport, which had been postulated to be present in RPE by others. We conclude that two processes are involved in regulation of pHi in RPE: A Na+/H+ antiport responsible for recovery of pHi from acid load, and a DIDS-sensitive Cl−/HCO −3 exchange mechanism responsible for recovery of pHi after alkalinization.

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References

  1. Akiba T, Alpern RJ, Eveloff J, Calamina J, Warnock DG (1986) Electrogenic sodium/bicarbonate cotransport in rabbit renal cortical basolateral membrane vesicles. J Clin Invest 78:1472–1478

    Google Scholar 

  2. Alpern RJ (1985) Mechanism of basolateral membrane H+/OH−/HCO −3 transport in the rat proximal convoluted tubule. J Gen Physiol 86:613–636

    Google Scholar 

  3. Aronson PS, Nee J, Suhm MA (1982) Modifier role of internal H+ in activating the Na+/H+-exchanger in renal microvillus membrane vesicles. Nature 299:161–163

    Google Scholar 

  4. Basu PK, Sarkar P, Menon I, Carre F, Persad S (1983) Bovine RPE cells cultured in vitro: growth characteristics, morphology, chromosomes, phagocytosis ability, tyrosinase activity and effect of freezing. Exp Eye Res 36:671–683

    Google Scholar 

  5. Benos DJ (1982) Amiloride: a molecular probe for sodium tansport in tissues and cells. Am J Physiol 242:C131-C145

    Google Scholar 

  6. Bok D (1982) Autoradiographic studies on the polarity of plasma membrane receptors in retinal pigment epithelial cells. In: Hollyfield J (ed) IVth International Symposium on the Structure of the Eye. Elsevier/North-Holland. Amsterdam New York, pp 247–256

    Google Scholar 

  7. Boron WF, and Boulpaep EL (1983) Intracellular pH regulation in the proximal tubule of the salamander kidney. Basolateral HCO −3 transport. J Gen Physiol 81:53–94

    Google Scholar 

  8. Boron WF, McCormick WC, Roos A (1979) pH regulation in barnacle muscle fibers: dependence on intracellular and extracellular pH. Am J Physiol 237:C185-C193

    Google Scholar 

  9. Chaillet JR, Boron WF (1985) Intracellular calibration of a pH-sensitive dye in isolated, perfused salamander proximal tubules. J Gen Physiol 86:765–794

    Google Scholar 

  10. Di Mattio J, Degnan KJ, Zadunaisky JA (1983) A model for transepithelial ion transport across the isolated retinal pigment epithelium of the frog. Exp Eye Res 37:409–420

    Google Scholar 

  11. Edwards RB (1977) Culture of rat retinal pigment epithelium. In Vitro 13:301–304

    Google Scholar 

  12. Frambach DA, Misfeldt DS (1983) Furosemide-sensitive Cl transport in embryonic chicken retinal pigment epithelium. Am J Physiol 244:F679-F685

    Google Scholar 

  13. Grassl SM, Aronson PS (1986) Na+/HCO −3 cotransport in basolateral membrane vesicles isolated from rabbit renal cortex. J Biol Chem 261:8778–8783

    Google Scholar 

  14. Jentsch TJ, Keller SK, Koch M, Wiederholt M (1984) Evidence for coupled transport of bicarbonate and sodium in cultured bovine corneal edothelial cells. J Membr Biol 81:189–204

    Google Scholar 

  15. Jentsch TJ, Stahlknecht TR, Hollwede H, Fischer DG, Keller SK, Wiederholt M (1985) A bicarbonate-dependent process inhibitable by disulfonic stilbenes and a Na+/H+ exchange mediate22Na+-uptake into cultured bovine corneal endothelium. J Biol Chem 260:795–801

    Google Scholar 

  16. Jentsch TJ, Janicke I, Sorgenfrei D, Keller SK, Wiederholt M (1986) The regulation of intracellular pH in monkey kidney epithelial cells (BSC-1). Roles of Na+/H+-antiport, Na+-HCO −3 -(NaCO −3 )-symport, and Cl−/HCO −3 -exchange. J Biol Chem 261:12120–12127

    Google Scholar 

  17. Jentsch TJ, Matthes H, Keller SK, Wiederholt M (1986) Electrical properties of sodium bicarbonate symport in kidney epithelial cells (BSC-1). Am J Physiol 251:F954-F968

    Google Scholar 

  18. Jentsch TJ, Korbmacher C, Fischer DG, Janicke I, Keller SK, Cragoe EJ, Wiederholt M (1987) The regulation of cytoplasmic pH of cultured bovine corneal endothelial cells in the absence and presence of bicarbonate. J Membr Biol (in press)

  19. Keller SK, Jentsch TJ, Koch M, Wiederholt M (1986) Interactions of pH and K+-conductance in cultured bovine retinal pigment epithelial cells. Am J Physiol 250:C124-C137

    Google Scholar 

  20. Keller SK, Jentsch TJ, Janicke I, Wiederholt M (1987) Na+/H+ antiport and Cl−/HCO −3 antiport in cultured bovine retinal pigment epithelium. Pflügers Arch 408 S1:R37 (Abstract)

    Google Scholar 

  21. Keller SK, Jentsch TJ, Wiederolt M (1986) Micropuncture and intracellular pH measurement in bovine retinal pigment epithelium in cell culture. Proc Int Soc for Eye Research IV:119 (Abstract)

    Google Scholar 

  22. Kinne-Saffran E, Kinne R, Rubinstein M, Zadunaisky J (1987) The existence of a Na+/H+ exchange mechanism in the apical membrane of the retinal pigment epithelium. Pflügers Arch 408 S1:R34 (Abstract)

    Google Scholar 

  23. L'Allemain G, Paris S, Pouyssegur J (1985) Role of Na+ dependent Cl−/HCO −3 exchange in regulation of intracellular pH in fibroblasts. J Biol Chem 260:4877–4883

    Google Scholar 

  24. Lasansky A, deFish FW (1966) Potential, current, and ionic fluxes across the isolated retinal pigment epithelium and choroid. J Gen Physiol 49:913–924

    Google Scholar 

  25. Miller SS, Steinberg RH (1982) Potassium transport across the frog retinal pigment epithelium. J Membr Biol 67:199–209

    Google Scholar 

  26. Miller SS, Steinberg RH, Oakley B II (1978) The electrogenic sodium pump of the frog retinal pigment epithelium. J Membr Biol 44:259–279

    Google Scholar 

  27. Montrose HM, Murer H (1986) Regulation of intracellular pH in LLC-PK1 cells by Na+/H+ exchange. J Membr Biol 93: 33–42

    Google Scholar 

  28. Ostwald TJ, Steinberg RH (1980) Localization of frog retinal pigment epithelium Na+-K+ ATPase. Exp Eye Res 31:351–360

    Google Scholar 

  29. Rosoff PM, Cantley LC (1985) Stimulation of the T3-T cell receptor-associated Ca2+ influx enhances the activity of the Na+/H+ exchanger in a leucemic human T-cell line. J Biol Chem 260:14053–14059

    Google Scholar 

  30. Rothenberg P, Glaser L, Schlesinger P, Cassel D (1983) Activation of Na+/H+-exchange by epidermal growth factor elevates intracellular pH in A431 cells. J Biol Chem 258:12644–12653

    Google Scholar 

  31. Simons ER, Schwarz DB, Norman E (1982) Stimulus response coupling in human platelets: Thrombin-induced changes in pH. In: Nuccitelli R, Deamer DW (eds) Intracellular pH: its measurement, regulation, and utilization in cellular functions. Liss, New York, pp 463–482

    Google Scholar 

  32. Steinberg RH, Miller SS (1979) Transport and membrane properties of the retinal pigment epithelium. In: Zinn KM, Marmor MF (eds) The retinal pigment epithelium. Harvard University Press, Boston, pp 205–225

    Google Scholar 

  33. Thomas JA, Buchsbaum RN, Zimniak A, Racker E (1979) Intracellular pH-measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. Biochem J 18:2210–2218

    Google Scholar 

  34. Thomas RC (1977) The role of bicarbonate, chloride and sodium ions in the regulation of intracellular pH in snail neurones. J Physiol 273:317–338

    Google Scholar 

  35. Wiederholt M, Zadunaisky JA (1984) Inhibition of intracellular chloride activity by furosemide in frog retinal pigment epithelium. Current Eye Res 3:673–675

    Google Scholar 

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Author notes

  1. Svea K. Keller

    Present address: Augenklinik und Poliklinik, Klinikum Steglitz, Preie Universität Berlin, Hindenburgdamm 30, D-1000, Berlin 45

  2. Thomas J. Jentsch

    Present address: Whitehead Institute for Biomedical Research, Nine Cambridge Center, 02142, Cambridge, MA, USA

Authors and Affiliations

  1. Institut für Klinische Physiologie, Klinikum Steglitz, Freie Universität Berlin, Hindenburgdamm 30, D-1000, Berlin 45

    Svea K. Keller, Thomas J. Jentsch, Ilse Janicke & Michael Wiederholt

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  1. Svea K. Keller
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  2. Thomas J. Jentsch
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  4. Michael Wiederholt
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Parts of this work jhave been published in abstract from [20, 21]

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Keller, S.K., Jentsch, T.J., Janicke, I. et al. Regulation of intracellular pH in cultured bovine retinal pigment epithelial cells. Pflugers Arch. 411, 47–52 (1988). https://doi.org/10.1007/BF00581645

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  • DOI: https://doi.org/10.1007/BF00581645

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