Pool depletion induces a novel calcium influx pathway activated by caffeine

C. A. Ufret-Vincenty, A. Alfpnso, Donald Gill

Research output: Contribution to journalArticle

Abstract

Co2+ influx through store-operated channels (SOCs) activated rapidly after Ca2+ pool depletion represents an important component of Ca2+ signals generated in cells. A new and distinct Ca2+ influx component activated by caffeine is induced in cells after Ca2+ pools are emptied using the intracellular Ca2+ pump inhibitors, thapsigargin (TG) or 2,5-di-fert- butylhydroquinone (DBHQ). Both blockers cause depletion of intracellular Ca2+ pools and eel! growth arrest; upon refilling of pools, normal cell-cycle progression is resumed (Short, A.D., et al. PNAS 90, 4986-4990, 1993). Here, the Co2+-sensitive dye, fura-2, was used to study Co2+ homeostasis in DDTMF - 2 smooth muscle cells growth-arrested by TG- or DBHQ-treatment. In DDTMF - 2 cells the SOC-mediated Ca2+ influx component after emptying Ca2+ pools is short- lived and appears to be rapidly deactivated. After treatment of DDTiMF - 2 cells with either 3 M TG or 10 /zM DBHQ, 10 mM caffeine induces a large transient influx of Ca2+ distinct from SOC- mediated Ca2+ entry. Caffeine-sensitive Ca2+ influx following DBHQ-treatment is activated more rapidly than that following TG-treatment. When caffeine is added to untreated DDT\MF - 2 cells no effect on cytosolic Co2+ concentration is observed. The disappearance of caffeine-induced Ca2+ influx is also different for TG- and DBHQ-treated cells. In DBHQ-treated cells, bradykinin- sensitive Ca2+ pools quickly refill and cells become insensitive to caffeine immediately after DBHQ removal. In the case of TG-Ueated cells, reversal of TG-induced growth arrest with either high (20%) serum or 1-10 /iM arachidonic acid, in addition to removal of TG, is required to allow agonist-sensitive Ca2+ pools to refill concomitantly with the disappearance of caffeine-induced Ca2+ influx. In summary, the results show that a Cu2+ influx pathway activated by caffeine is observed under conditions of growth arrest induced by either TG or DBHQ and appears to be directly correlated with depletion of intracellular Ca2+ pools. (NIH grants NS19304 and GM15407; NSF grant MCB 9307746).

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - Dec 1 1996

Fingerprint

Thapsigargin
caffeine
Caffeine
Calcium
calcium
Organized Financing
Cells
Growth
cells
Eels
DDT
Cell growth
Bradykinin
Arachidonic Acid
Smooth Muscle Myocytes
Muscle
Cell Cycle
Homeostasis
Coloring Agents
bradykinin

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Ufret-Vincenty, C. A. ; Alfpnso, A. ; Gill, Donald. / Pool depletion induces a novel calcium influx pathway activated by caffeine. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
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title = "Pool depletion induces a novel calcium influx pathway activated by caffeine",
abstract = "Co2+ influx through store-operated channels (SOCs) activated rapidly after Ca2+ pool depletion represents an important component of Ca2+ signals generated in cells. A new and distinct Ca2+ influx component activated by caffeine is induced in cells after Ca2+ pools are emptied using the intracellular Ca2+ pump inhibitors, thapsigargin (TG) or 2,5-di-fert- butylhydroquinone (DBHQ). Both blockers cause depletion of intracellular Ca2+ pools and eel! growth arrest; upon refilling of pools, normal cell-cycle progression is resumed (Short, A.D., et al. PNAS 90, 4986-4990, 1993). Here, the Co2+-sensitive dye, fura-2, was used to study Co2+ homeostasis in DDTMF - 2 smooth muscle cells growth-arrested by TG- or DBHQ-treatment. In DDTMF - 2 cells the SOC-mediated Ca2+ influx component after emptying Ca2+ pools is short- lived and appears to be rapidly deactivated. After treatment of DDTiMF - 2 cells with either 3 M TG or 10 /zM DBHQ, 10 mM caffeine induces a large transient influx of Ca2+ distinct from SOC- mediated Ca2+ entry. Caffeine-sensitive Ca2+ influx following DBHQ-treatment is activated more rapidly than that following TG-treatment. When caffeine is added to untreated DDT\MF - 2 cells no effect on cytosolic Co2+ concentration is observed. The disappearance of caffeine-induced Ca2+ influx is also different for TG- and DBHQ-treated cells. In DBHQ-treated cells, bradykinin- sensitive Ca2+ pools quickly refill and cells become insensitive to caffeine immediately after DBHQ removal. In the case of TG-Ueated cells, reversal of TG-induced growth arrest with either high (20{\%}) serum or 1-10 /iM arachidonic acid, in addition to removal of TG, is required to allow agonist-sensitive Ca2+ pools to refill concomitantly with the disappearance of caffeine-induced Ca2+ influx. In summary, the results show that a Cu2+ influx pathway activated by caffeine is observed under conditions of growth arrest induced by either TG or DBHQ and appears to be directly correlated with depletion of intracellular Ca2+ pools. (NIH grants NS19304 and GM15407; NSF grant MCB 9307746).",
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Pool depletion induces a novel calcium influx pathway activated by caffeine. / Ufret-Vincenty, C. A.; Alfpnso, A.; Gill, Donald.

In: FASEB Journal, Vol. 10, No. 6, 01.12.1996.

Research output: Contribution to journalArticle

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T1 - Pool depletion induces a novel calcium influx pathway activated by caffeine

AU - Ufret-Vincenty, C. A.

AU - Alfpnso, A.

AU - Gill, Donald

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Y1 - 1996/12/1

N2 - Co2+ influx through store-operated channels (SOCs) activated rapidly after Ca2+ pool depletion represents an important component of Ca2+ signals generated in cells. A new and distinct Ca2+ influx component activated by caffeine is induced in cells after Ca2+ pools are emptied using the intracellular Ca2+ pump inhibitors, thapsigargin (TG) or 2,5-di-fert- butylhydroquinone (DBHQ). Both blockers cause depletion of intracellular Ca2+ pools and eel! growth arrest; upon refilling of pools, normal cell-cycle progression is resumed (Short, A.D., et al. PNAS 90, 4986-4990, 1993). Here, the Co2+-sensitive dye, fura-2, was used to study Co2+ homeostasis in DDTMF - 2 smooth muscle cells growth-arrested by TG- or DBHQ-treatment. In DDTMF - 2 cells the SOC-mediated Ca2+ influx component after emptying Ca2+ pools is short- lived and appears to be rapidly deactivated. After treatment of DDTiMF - 2 cells with either 3 M TG or 10 /zM DBHQ, 10 mM caffeine induces a large transient influx of Ca2+ distinct from SOC- mediated Ca2+ entry. Caffeine-sensitive Ca2+ influx following DBHQ-treatment is activated more rapidly than that following TG-treatment. When caffeine is added to untreated DDT\MF - 2 cells no effect on cytosolic Co2+ concentration is observed. The disappearance of caffeine-induced Ca2+ influx is also different for TG- and DBHQ-treated cells. In DBHQ-treated cells, bradykinin- sensitive Ca2+ pools quickly refill and cells become insensitive to caffeine immediately after DBHQ removal. In the case of TG-Ueated cells, reversal of TG-induced growth arrest with either high (20%) serum or 1-10 /iM arachidonic acid, in addition to removal of TG, is required to allow agonist-sensitive Ca2+ pools to refill concomitantly with the disappearance of caffeine-induced Ca2+ influx. In summary, the results show that a Cu2+ influx pathway activated by caffeine is observed under conditions of growth arrest induced by either TG or DBHQ and appears to be directly correlated with depletion of intracellular Ca2+ pools. (NIH grants NS19304 and GM15407; NSF grant MCB 9307746).

AB - Co2+ influx through store-operated channels (SOCs) activated rapidly after Ca2+ pool depletion represents an important component of Ca2+ signals generated in cells. A new and distinct Ca2+ influx component activated by caffeine is induced in cells after Ca2+ pools are emptied using the intracellular Ca2+ pump inhibitors, thapsigargin (TG) or 2,5-di-fert- butylhydroquinone (DBHQ). Both blockers cause depletion of intracellular Ca2+ pools and eel! growth arrest; upon refilling of pools, normal cell-cycle progression is resumed (Short, A.D., et al. PNAS 90, 4986-4990, 1993). Here, the Co2+-sensitive dye, fura-2, was used to study Co2+ homeostasis in DDTMF - 2 smooth muscle cells growth-arrested by TG- or DBHQ-treatment. In DDTMF - 2 cells the SOC-mediated Ca2+ influx component after emptying Ca2+ pools is short- lived and appears to be rapidly deactivated. After treatment of DDTiMF - 2 cells with either 3 M TG or 10 /zM DBHQ, 10 mM caffeine induces a large transient influx of Ca2+ distinct from SOC- mediated Ca2+ entry. Caffeine-sensitive Ca2+ influx following DBHQ-treatment is activated more rapidly than that following TG-treatment. When caffeine is added to untreated DDT\MF - 2 cells no effect on cytosolic Co2+ concentration is observed. The disappearance of caffeine-induced Ca2+ influx is also different for TG- and DBHQ-treated cells. In DBHQ-treated cells, bradykinin- sensitive Ca2+ pools quickly refill and cells become insensitive to caffeine immediately after DBHQ removal. In the case of TG-Ueated cells, reversal of TG-induced growth arrest with either high (20%) serum or 1-10 /iM arachidonic acid, in addition to removal of TG, is required to allow agonist-sensitive Ca2+ pools to refill concomitantly with the disappearance of caffeine-induced Ca2+ influx. In summary, the results show that a Cu2+ influx pathway activated by caffeine is observed under conditions of growth arrest induced by either TG or DBHQ and appears to be directly correlated with depletion of intracellular Ca2+ pools. (NIH grants NS19304 and GM15407; NSF grant MCB 9307746).

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