title: Calcium signaling in human airway goblet cells following purinergic activation.
abstract: Despite the general importance of Ca(2+) signaling in signal transduction, and of goblet cell mucin hypersecretion in inflammatory pulmonary diseases, measurement of airway goblet cell intracellular Ca(2+) (Ca(i)(2+)) has not been reported. In this article, we describe the results of experiments measuring Ca(i)(2+) in primary cultures of human bronchial goblet cells after stimulation with the purinergic agonist adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS) and phorbol 12-myristate 13-acetate (PMA). Ca(2+) signaling in human goblet cells after purinergic stimulation follows the classic paradigm of a Ca(i)(2+) transient from a basal activity of 110 nM to a peak response of 260.1 +/- 41.2 nM within 2 min, followed by a long superbasal plateau (155.3 +/- 0.2 nM) between 10 and 15 min. The rise in Ca(i)(2+) appears to result from a mobilization of intracellular stores, because the transient was nearly abolished by inhibition of PLC with the phosphatidylinositol-specific PLC inhibitor U-73122, and it was not affected significantly by removal of extracellular Ca(2+). Loading goblet cells with BAPTA inhibited the ATPgammaS-induced Ca(2+) transient by 86.0 +/- 13.1%, relative to control. Finally, in contrast to the massive effects of high doses of PMA (300 nM) on mucin secretion from goblet cells, phorbol ester stimulated a small (27.1 +/- 7% of the ATPgammaS control peak), brief rise in Ca(i)(2+). This diminutive signal likely denotes a local Ca(2+) gradient, which may be associated with the mucin granule exocytotic process.
authors.affiliation: CF/Pulmonary Research & Treatment Center, Department of Cell and Molecular Physiology, University of North Carolina, Chapel Hill, NC 27599-7248, USA.
authors.forename: Andrea H
authors.initials: AH
authors.lastname: Rossi
authors.forename: Wendy C
authors.initials: WC
authors.lastname: Salmon
authors.forename: Michael
authors.initials: M
authors.lastname: Chua
authors.forename: C William
authors.initials: CW
authors.lastname: Davis
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D001952:Bridged-Ring Compounds / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D020013:Calcium Signaling / Q000187:drug effects*; D002478:Cells, Cultured; D004963:Estrenes / Q000494:pharmacology; D020397:Goblet Cells / Q000187:drug effects* / Q000502:physiology*; D006801:Humans; D009636:Norbornanes; D043265:Phosphatidylinositol Diacylglycerol-Lyase / Q000037:antagonists & inhibitors; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D058910:Purinergic P2 Receptor Agonists; D011760:Pyrrolidinones / Q000494:pharmacology; D058473:Receptors, Purinergic P2Y2; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors
chemical_list: D001952:Bridged-Ring Compounds; D004963:Estrenes; D009636:Norbornanes; C546385:P2RY2 protein, human; D010726:Phosphodiesterase Inhibitors; D058910:Purinergic P2 Receptor Agonists; D011760:Pyrrolidinones; D058473:Receptors, Purinergic P2Y2; D013859:Thiocarbamates; D013871:Thiones; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C022571:adenosine 5'-O-(3-thiotriphosphate); C046498:tricyclodecane-9-yl-xanthogenate; D000255:Adenosine Triphosphate; D010738:Type C Phospholipases; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D013755:Tetradecanoylphorbol Acetate