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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

Abstract

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 

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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

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