title: Modulation of CCK-8-evoked intracellular Ca2+ waves by hydrogen peroxide in mouse pancreatic acinar cells.
abstract: In the present study we have employed single cell imaging analysis to monitor the propagation of cholecystokinin-evoked Ca(2+) waves in mouse pancreatic acinar cells. Stimulation of cells with 1 nM CCK-8 led to an initial Ca(2+) release at the luminal cell pole and subsequent spreading of the Ca(2+) signal towards the basolateral membrane in the form of a Ca(2+) wave. Inhibition of sarcoendoplasmic reticulum Ca(2+)-ATPase (SERCA) activity by 1 microM thapsigargin, preincubation in the presence of 100 microM H(2)O(2) or inhibition of PKC with either 5 microM Ro31-8220 or 3 microM GF-109203-X all led to a faster propagation of CCK-8-induced Ca(2+) signals. The propagation of CCK-8-evoked Ca(2+) signals was slowed down by activation of PKC with 1 microM PMA, and preincubation of cells in the presence of H(2)O(2) counteracted the effect of PKC inhibition. The protonophore FCCP (100 nM) and the inhibitor of the mitochondrial Ca(2+)-uniporter Ru360 (10 microM) led to an increase in the propagation rate of CCK-8-evoked Ca(2+) waves. Finally, depolymerisation of actin cytoskeleton with cytochalasin D (10 microM) led to a faster propagation of CCK-8-evoked Ca(2+) signals. Stabilization of actin cytoskeleton with jasplakinolide (10 microM) did not induce significant changes on CCK-8-evoked Ca(2+) waves. Preincubation of cells in the presence of H(2)O(2) counteracted the effect of cytochalasin D on CCK-8-evoked Ca(2+) wave propagation. Our results suggest that spreading of cytosolic Ca(2+) waves evoked by CCK-8 can be modulated by low levels of oxidants acting on multiple Ca(2+)-handling mechanisms.
authors.affiliation: Department of Physiology (Cell Physiology Research Group), University of Extremadura. Cáceres, Spain.
authors.forename: M P
authors.initials: MP
authors.lastname: Granados
authors.forename: G M
authors.initials: GM
authors.lastname: Salido
authors.forename: J A
authors.initials: JA
authors.lastname: Pariente
authors.forename: A
authors.initials: A
authors.lastname: Gonzáles
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects*; D002232:Carbocyanines / Q000494:pharmacology; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002766:Cholecystokinin / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D047630:Depsipeptides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology; D007424:Intracellular Fluid / Q000187:drug effects / Q000502:physiology; D008297:Male; D008301:Maleimides / Q000494:pharmacology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D009840:Oligomycins / Q000494:pharmacology; D009947:Organotin Compounds / Q000494:pharmacology; D046790:Pancreas, Exocrine / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D017975:Ruthenium Compounds / Q000494:pharmacology; D012459:Salicylates / Q000494:pharmacology; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism; D012844:Sincalide / Q000494:pharmacology*; D019284:Thapsigargin / Q000494:pharmacology
chemical_list: D001562:Benzimidazoles; D002232:Carbocyanines; D047630:Depsipeptides; D007211:Indoles; D008301:Maleimides; D009840:Oligomycins; D009947:Organotin Compounds; C112020:Ru 360; D017975:Ruthenium Compounds; D012459:Salicylates; C417088:triphenyltin salicylate; C057531:jasplakinolide; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; D015638:Cytochalasin D; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D019284:Thapsigargin; D002766:Cholecystokinin; D006861:Hydrogen Peroxide; D011493:Protein Kinase C; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; C070515:bisindolylmaleimide I; D012844:Sincalide; C064758:Ro 31-8220