Pflügers Archiv

, Volume 427, Issue 1–2, pp 33–41 | Cite as

Activation of calcium influx by ATP and store depletion in primary cultures of renal proximal cells

  • Jean -Christophe Cejka
  • Sophie Le Maout
  • Michel Bidet
  • Michel Tauc
  • Philippe Poujeol
Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands


Cytoplasmic calcium changes and calcium influx evoked by adenosine triphosphate (ATP) were investigated in primary cultures of rabbit proximal convoluted tubule cells. Extracellular ATP (50 μM) induced a biphasic increase of [Ca2+]i measured with the calcium probe fura-2. In the early phase, the mobilization of intracellular pools resulted in a transient increase of [Ca2+]i from 106±11 nM (n=36) to 1059±115% (n=29) of the resting level within 10 s. In the presence of external calcium, [Ca2+]i then decreased within 3 min to a sustained level (398±38%,n=8). Measurements of fura-2 quenching by external manganese revealed that this phase was the result of an increased Ca2+ uptake, blocked by lanthanum (10 μM) and verapamil (100 μM) but not by the nifedipin (25 μM). Internal calcium store depletion by ATP induced an increased calcium influx through lanthanum- and verapamil-sensitive, nifedipininsensitive calcium channels, located on the apical membrane of the cells. As indicated by86Rb+ efflux measurements, ATP activated a potassium efflux that was blocked by barium andLeiurus quinquestriatus hebraeus (LQH) venom (containing charybdotoxin) indicating the involvement of Ca2+-sensitive K+ channels. Moreover, in the presence of the LQH venom, the internal calcium stores were not replenished after being depleted by ATP. Our results indicate that an ATPevoked hyperpolarization of the plasma membrane leads to increased Ca2+ influx, which facilitates the replenishment of the internal stores.

Key words

Renal proximal tubule Intracellular calcium Calcium influx Primary culture Calcium store depletion 86Rb efflux Calcium sensitive potassium channels 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Backsai BJ, Friedman PA (1990) Activation of latent Ca2+ channels in renal epithelial cells by parathyroid hormone. Nature 347:388–391Google Scholar
  2. 2.
    Bidet M, Tauc M, Koechlin N, Poujeol P (1990) Video microscopy of intracellular pH in primary cultures of rabbit proximal and early distal tubules. Pflügers Arch 416:270–280Google Scholar
  3. 3.
    Cejka JC, Bidet M, Tauc M, Poujeol P (1993) Nucleotides mobilize intracellular calcium stores of renal proximal cells in primary culture: existence of a suramin-sensitive mechanism. Biochim Biophys Acta 1176:7–12Google Scholar
  4. 4.
    Dominguez JH, Mann C, Rothrock JK, Bhati V (1991) Na+- Ca+ exchange and Ca2+ depletion in rat proximal tubules. Am J Physiol 261:F328-F335Google Scholar
  5. 5.
    Doucet A, Katz AI (1982) High-affinity Ca-Mg-ATPase along the rabbit nephron. Am J Physiol 252:F346-F352Google Scholar
  6. 6.
    Garcia ML, Galvez A, Garcia-Calvo M, King VF, Vaszquez J, Kaczorowsky GJ (1991) Use of toxins to study potassium channels. J Bioenerg Biomembr 23:4Google Scholar
  7. 7.
    Gmaj P, Murer H (1988) Calcium transport mechanisms in epithelial cell membranes. Miner Electrolyte Metab 14:22–30Google Scholar
  8. 8.
    Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450Google Scholar
  9. 9.
    Hallam TJ, Rink TJ (1985) Agonists stimulate divalent cation channels in the plasma membrane of human platelets. FEBS Lett 186:175–179Google Scholar
  10. 10.
    Hallam TJ, Rink TJ (1989) Receptor-mediated Ca2+ entry: diversity of function and mechanism. Trends Pharmacol Sci 10:8–10Google Scholar
  11. 11.
    Hoth M, Penner R (1992) Depletion on intracellular calcium stors activates a calcium current in mast cells. Nature 355:353–356Google Scholar
  12. 12.
    Jacob R (1990) Agonist-stimulated divalent cation entry into single cultured human umbilical vein endothelial cells. J Physiol (Lond) 421:55–57Google Scholar
  13. 13.
    Jacobsen C, Roigaard-Petersen H, Sheikh MI (1989) Ba2+- sensitive K+ channels in luminal-membrane vesicles from pars convoluta of rabbit proximal tubule. FEBS Lett 243:173–176Google Scholar
  14. 14.
    Kinne-Saffran E, Kinne R (1974) Localization of a calciumstimulated ATPase in the basal-lateral plasma membrane of the proximal tubule of rat kidney cortex. J Membr Biol 17:263–274Google Scholar
  15. 15.
    Le Maout S, Tauc M, Koechlin K, Poujeol P (1990) Polarized86Rb+ effluxes in primary cultures of rabbit kidney proximal cells: role of calcium and hypotonicity. Biochim Biophys Acta 1026:29–39Google Scholar
  16. 16.
    Mérot J, Bidet M, Gachot B, Le Maout S, Tauc M, Poujeol P (1988) Patch clamp study on primary culture of isolated proximal convoluted tubules. Pflügers Arch 413:51–61Google Scholar
  17. 17.
    Merritt JE, Jacob R, Hallam TJ (1989) Use of manganese to discriminate between calcium influx and mobilization from internal stores in stimulated human neutrophils. J Biol Chem 264:1522–1587Google Scholar
  18. 18.
    Paulmichl M, Lang F (1988) Enhancement of intracellular calcium concentration by extracellular ATP and UTP in Madin Darby canine kidney cells. Biochem Biophys Res Commun 156:1139–1143Google Scholar
  19. 19.
    Poncet V, Merot J, Poujeol P (1992) A calcium-permeable channel in the apical membrane of primary cultures of the rabbit distal bright convoluted tubule. Pflügers Arch 422:112–119Google Scholar
  20. 20.
    Suki WN, Rouse D (1991) Renal transport of calcium, magnesium, and phosphorus. In: WB Saunders Company, Philadelphia (ed) The kidney, 4th edn. Elements of normal renal function, vol I. Chapter 10, pp 380–423Google Scholar
  21. 21.
    Tauc M, Merot J, Bidet M, Koechlin N, Gastineau M, Othmani L, Poujeol P (1989) Antigenic expression of aminopeptidase M, dipeptidyl-peptidase IV and endopeptidase by primary cultures from rabbit kidney proximal tubule. Histochemistry 91:17–30Google Scholar
  22. 22.
    Tauc M, Gastineau M, Poujeol P (1992) Toxin pharmacology of the ATP-induced hyperpolarization in Madin-Darby canine kidney cells. Biochim Biophys Acta 1105:155–160Google Scholar
  23. 23.
    Tsien RW, Tsien RY (1990) Calcium channels, stores and oscillations. Annu Rev Cell Biol 6:715–760Google Scholar
  24. 24.
    Ullrich KJ, Rumrich G, Klöss S (1976) Active Ca2+ reabsorption in the proximal tubule of the rat kidney-dependence on sodium and buffer transport. Pflügers Arch 364:223–228Google Scholar
  25. 25.
    Van Heeswijck MPE, Geertsen JAM, Van Os CH (1984) Kinetic properties of the ATP-dependent Ca2+ pump and the Na+/ Ca2+ exchange system in basolateral membranes from rat kidney cortex. J Membr Biol 79:19–31Google Scholar
  26. 26.
    Vieyra A, Nachbin L, Dios-Abad E de, Goldfeld M, Meyer-Fernandes JR, Moares L de (1986) Comparison between calcium transport and adenosine triphosphate activity in membrane vesicles derived from rabbit kidney proximal tubule. J Biol Chem 261:4247–4255Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Jean -Christophe Cejka
    • 1
  • Sophie Le Maout
    • 1
  • Michel Bidet
    • 1
  • Michel Tauc
    • 1
  • Philippe Poujeol
    • 1
  1. 1.Département de Biologie Cellulaire et Moléculaire, Service de Biologie CellulaireCentre d'Etudes de SaclayGif sur YvetteFrance

Personalised recommendations