Activation of calcium influx by ATP and store depletion in primary cultures of renal proximal cells.
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 microM) 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 microM) and verapamil (100 microM) but not by the nifedipin (25 microM). Internal calcium store depletion by ATP induced an increased calcium influx through lanthanum- and verapamil-sensitive, nifedipin-insensitive calcium channels, located on the apical membrane of the cells. As indicated by 86Rb+ efflux measurements, ATP activated a potassium efflux that was blocked by barium and Leiurus quinquestriatus hebraeus (LQH) venom (containing charybdotoxin) indicating the involvement of Ca(2+)-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 ATP-evoked hyperpolarization of the plasma membrane leads to increased Ca2+ influx, which facilitates the replenishment of the internal stores.
Cejka
J C
JC
Département de Biologie Cellulaire et Moléculaire, Centre d'Etudes de Saclay, Gif sur Yvette, France.
Le Maout
S
S
Bidet
M
M
Tauc
M
M
Poujeol
P
P
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000172:deficiency / Q000378:metabolism*; D016764:Cell Polarity; D002478:Cells, Cultured; D018999:Charybdotoxin; D005110:Extracellular Space / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D011188:Potassium / Q000378:metabolism; D011817:Rabbits; D012604:Scorpion Venoms / Q000494:pharmacology
D012604:Scorpion Venoms; D018999:Charybdotoxin; D000255:Adenosine Triphosphate; D011188:Potassium; D002118:Calcium