Pflügers Archiv

, Volume 428, Issue 1, pp 60–68 | Cite as

Volume regulatory responses in frog isolated proximal cells

  • L. Robson
  • M. Hunter
Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands


Cells respond to increases in volume by activating solute efflux pathways, resulting in water loss and restoration of the original cell volume. The solute efflux pathways underlying these volume regulatory decrease (VRD) responses have been relatively well studied. However, the transduction pathways whereby the change in cell volume is converted into an intracellular signal resulting in VRD are much less well understood. We have examined VRD in isolated proximal tubule cells from the frog, with particular attention to the roles of stretch-activated channels, Ca2+ and protein kinases. Cell length was taken as an index of cell volume, and was measured continuously using a photodiode array. VRD was observed in approximately 50% of cells, and was inhibited by Ba2+, Gd3+ and 4,4′-diisothiocyanatostilbene 2,2′-disulphonic acid (DIDS), and removal of extracellular Ca2+. VRD was accelerated by the active phorbol ester, phorbol 12-myristate 13-acetate (PMA), and the phosphatase inhibitor F; on the other hand, VRD was prolonged by 4α-phorbol 12,13-didecanoate (PDC), an inactive phorbol ester), and inhibited by PMA and Gd3+, PMA and 0 Ca2+, and staurosporine. Volume regulation was unaffected by di-butyryl cAMP and 3-isobutyl-1-methyl-xanthene (IBMX). These data suggest that Ca2+ and PKC, via protein phosphorylation, play a stimulatory role in VRD.

Key words

Volume regulation Calcium Barium DIDS Stretch-activated channel Protein kinase C 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Beck JS, Breton S, Laprade R, Giebisch G (1991) Volume regulation and intracellular calcium in the rabbit proximal convoluted tubule. Am J Physiol 260:F861-F867Google Scholar
  2. 2.
    Boyett MR, Moore M, Jewell BR, Montgomery RAP, Kirby MS, Orchard CH (1988) An improved apparatus for the optical recording of contraction of single heart cells. Pflügers Arch 413:197–205Google Scholar
  3. 3.
    Christensen O (1987) Mediation of cell volume regulation by calcium influx through stretch-activated channels. Nature 330:66–68Google Scholar
  4. 4.
    Cohen P (1988) Protein phosphorylation and hormone action. Proc R Soc Lond [Biol] 234:115–144Google Scholar
  5. 5.
    Filipovic D, Sackin H (1991) A calcium-permeable stretch activated cation channel in renal proximal tubule. Am J Physiol 260:f119-f129Google Scholar
  6. 6.
    Filipovic D, Sackin H (1992) Stretch-and volume-activated channels in isolated proximal tubule cells. Am J Physiol 262:F857-F870Google Scholar
  7. 7.
    Germann WJ, Ernst SA, Dawson DC (1986) Resting and osmotically induced basolateral potassium conductances in turtle colon. J Gen Physiol 88:225–237Google Scholar
  8. 8.
    Gonzalez E, Carpi-Medina P, Whittembury G (1982) Cell osmotic water permeability of isolated rabbit proximal straight tubules. Am J Physiol 242:F321-F330Google Scholar
  9. 9.
    Grinstein S, Cohen S, Goetz JD, Rothstein A, Mellors A, Gelfand EW (1986) Activation of the Na+-H+ antiport by changes in cell volume and by phorbol esters; possible role of protein kinase. Current Top Membr Transp 26:115–134Google Scholar
  10. 10.
    Grinstein S, Furuya W, Bianchini L (1992) Protein kinases, phosphatases, and the control of cell volume. News Physiol Sci 7:232–237Google Scholar
  11. 11.
    Grinstein S, Smith JD (1990) Calcium-independent cell volume regulation in human lymphocytes. J Gen Physiol 95:97–120Google Scholar
  12. 12.
    Haussinger D, Hallbrucker C, Vom Dahl S, Decker S, Schwiezer U, Lang F, Gerok W (1991) Cell volume is a major determinant of proteolysis control in liver. FEBS Lett 283:70–72Google Scholar
  13. 13.
    Hoffmann EK, Simonsen LO (1989) Membrane mechanisms in volume and pH regulation in vertebrate cells. Physiol Rev 69:315–382Google Scholar
  14. 14.
    House CR (1974) In: Water transport in cells and tissues. Arnold, pp 193–262Google Scholar
  15. 15.
    Hunter M (1989) Isolation of single proximal cells from frog kidneys. J Physiol (Lond) 416:13PGoogle Scholar
  16. 16.
    Hunter M (1990) Stretch-activated channels in the basolateral membrane of single proximal cells of frog kidney. Pflügers Arch 416:448–453Google Scholar
  17. 17.
    Hunter M (1991) Potassium-selective channels in the basolateral membrane of single proximal cells of frog kidney. Pflügers Arch 418:26–34Google Scholar
  18. 18.
    Hwang TK, Lu L, Zeitlin PL, Gruenert DC, Huganir R, Guggino WB (1989) Cl channels in CF: lack of activation by protein kinase C and cAMP-dependent protein kinase. Science 244:1351–1353Google Scholar
  19. 19.
    Jennings ML, Schulz RK (1991) Okadaic acid inhibition of KCl cotransport. Evidence that protein dephosphorylation is necessary for activation of transport by either cell swelling or N-ethylmaleimide. J Gen Physiol 97:799–818Google Scholar
  20. 20.
    Kaji DM, Tsukitani Y (1991) Role of protein phosphatase in activation of KCl cotransport in human erythrocytes. Am J Physiol 260:C176-C182Google Scholar
  21. 21.
    Kawahara K (1990) A stretch-activated K channel in the basolateral membrane of Xenopus kidney proximal tubule cells. Pflügers Arch 415:624–629Google Scholar
  22. 22.
    Kirk KL, DiBona DR, Schafer JA (1987) Regulatory volume decrease in perfused proximal nephron: evidence for a dumping of cell potassium. Am J Physiol 252:F933-F942Google Scholar
  23. 23.
    Kirk KL, Schafer JA, DiBona DR (1987) Cell volume regulation in rabbit proximal straight tubule perfused in vitro. Am J Physiol 252:F922-F932Google Scholar
  24. 24.
    Lang F, Oberleithner H, Giebisch G (1986) Electrophysiological heterogeneity of proximal convoluted tubules in Amphiuma kidney. Am J Physiol 251:F1063-F1072Google Scholar
  25. 25.
    Li M, McCann JD, Anderson MP, Clancy JP, Liedtke CM, Nairn AC, Greengard P, Welsh MJ (1989) Regulation of chloride channels by protein kinase C in normal and cystic fibrosis airway epithelia. Science 244:1353–1356Google Scholar
  26. 26.
    Lopes AG, Guggino WB (1987) Volume regulation in the early proximal tubule of the Necturus kidney. J Membr Biol 97:117–125Google Scholar
  27. 27.
    McCarty NA, O'Neil RG (1991) Calcium-dependent control of volume regulation in renal proximal tubule cells: 1. Swelling-activated Ca2+ entry and release. J Membr Biol 123:149–160Google Scholar
  28. 28.
    Mitsuka M, Berk BC (1991) Long-term regulation of Na+-H+ exchange in vascular smooth muscle cells: role of protein kinase C. Am J Physiol 260:C562-C569Google Scholar
  29. 29.
    Montrose-Rafizadeh C, Guggino WB (1990) Cell volume regulation in the nephron. Annu Rev Physiol 52:761–772Google Scholar
  30. 30.
    Nishizuka Y (1984) The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature 308:693–698Google Scholar
  31. 31.
    Parker JC, Colclasure GC, McManus TJ (1991) Coordinated regulation of shrinkage-induced Na/H exchange and swelling induced [K-Cl] cotransport in dog red cells. J Gen Physiol 98:869–880Google Scholar
  32. 32.
    Ribalet B, Eddlestone GT, Ciani S (1988) Metabolic regulation of the K(ATP) and a maxi-K(V) channel in the insulinsecreting RINm5F cell. J Gen Physiol 92:219–237Google Scholar
  33. 33.
    Roy G, Sauve R (1987) Effect of anisotonic media on volume, ion and amino-acid content and membrane potential of kidney cells (MDCK) in culture. J Membr Biol 100:83–96Google Scholar
  34. 34.
    Sackin H (1987) Stretch-activated potassium channels in renal proximal tubule. Am J Physiol 253:F1253-F1262Google Scholar
  35. 35.
    Siebens AW (1985) Cellular volume control. In: Seidin DW, Giebisch G (eds) The Kidney: physiology and pathophysiology. Raven, New York pp 91–115Google Scholar
  36. 36.
    Spring KR, Giebisch G (1977) Kinetics of Na transport in Necturus proximal tubule. J Gen Physiol 70:307–328Google Scholar
  37. 37.
    Suzuki M, Kawahara K, Ogawa A, Morita T, Kawaguchi Y, Kurihara S, Sakai O (1990) Intracellular calcium rises via G protein during regulatory volume decrease in rabbit proximal tubule cells. Am J Physiol 285:F690-F696Google Scholar
  38. 38.
    Tojyo Y, Tanimura A, Matsui S, Matsumoto Y, Sugiya H, Furuyama S (1991) NaF-induced amylase release from rat parotid cells is mediated by PI breakdown leading to Ca2+ mobilization. Am J Physiol 260:C194-C200Google Scholar
  39. 39.
    Ubl J, Murer H, Kolb HA (1988) Ion channels activated by osmotic and mechanical stress in membranes of opossum kidney cells. J Membr Biol 104:223–232Google Scholar
  40. 40.
    Volkl H, Lang F (1988) Ionic requirements for regulatory cell volume decrease in renal straight tubules (proximal). Pflügers Arch 412:1–6Google Scholar
  41. 41.
    Wang W, Giebisch G (1991) Dual modulation of renal ATP-sensitive K-channel by protein kinase A and C. Proc Natl Acad Sci USA 88:9722Google Scholar
  42. 42.
    Welling PA, Linshaw MA (1988) Importance of anion in hypotonic volume regulation in rabbit straight tubule. Am J Physiol 255:F853-F860Google Scholar
  43. 43.
    Welling PA, O'Neil RG (1990) Cell swelling activates basolateral membrane Cl and K conductances in rabbit proximal tubule. Am J Physiol 258:F951-F962Google Scholar
  44. 44.
    Welling PA, Linshaw MA, Sullivan LP (1985) Effect of Ba on cell volume regulation in rabbit proximal straight tubules. Am J Physiol 249:F20-F27Google Scholar
  45. 45.
    Whittembury G, Lindemann B, Carpi-Medina P, Gonzalez E, Linares H (1986) Continuous measurements of cell volume changes in single kidney tubules. Kidney Int 30:187–191Google Scholar
  46. 46.
    Yang X, Sachs F (1989) Block of stretch-activated ion channels in Xenopus oocytes by gadolinium and calcium ions. Science 243:1068–1071Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • L. Robson
    • 1
  • M. Hunter
    • 1
  1. 1.Department of Physiology, The Medical SchoolLeeds UniversityLeedsUK

Personalised recommendations