A novel methylxanthine-sensmve ca2+ influx mechanism is dependent on ca2+ pool emptying

Carmen A. Ufret-Vincentv, Donald Gill

Research output: Contribution to journalArticle

Abstract

Ca2+ influx through store-operated channels (SOCs) activated by Ca21 pool depletion represents an important component of Ca2+ signals generated in cells. We described a new and distinct Ca2+ influx component induced in cells after Ca2+ pools are emptied with thapsigargin (TG) (Ufret-Vincenty, C.A., et al., JBC 270, 26790-26793, 1995). This novel influx pathway can be activated by caffeine and other methylxanthines. We have further characterized this novel influx mechanism in DDT,MF-2 smooth muscle cells. The SOC-mediated Ca2+ influx component after emptying Ca2+ pools in DDT,MF-2 cells was short-lived and appeared to be rapidly deactivated. After treatment of DDT,MF-2 cells with 10 nM of the Ca2+locker, 2,5-di-fert-butylhydroquinone (DBHQ), 10 mM caffeine induced a rapid, large, but transient influx of Ca2+. This influx was distinct from SOC-mediated Ca2+ entry. Caffeine added to untreated DDT,MF-2 cells had no effect on cytosolic Ca2+ concentration, thus the caffeine-induced Ca2+ influx is active solely in the pool-depleted state. In DBHQ-treated cells, bradykinin-sensitive Ca 2+ pools quickly refilled and cells became insensitive to caffeine immediately after DBHQ removal. With TG-treated cells, reversal of TG-induced pool depletion required a high (20%) serum treatment, in addition to removal of TG, to allow refilling of agonist-sensitive Ca2+ pools concomitantly with the disappearance of caffeine-induced Ca2+ influx. Caffeine-activated Ca2+ influx was also induced by emptying intracellular pools using the Ca2+ ionophpre, ionomycin (10 nM), suggesting that emptying intracellular Ca2+ stores induces caffeine sensitivity, regardless of the means used for depletion. The results show that the appearance of caffeine-sensitive Ca2+ influx is directly correlated with depletion of intracellular Ca2+ pools.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - Dec 1 1997

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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