title: Effect of thapsigargin on cytosolic Ca2+ level and amylase release in rat parotid acinar cells.
abstract: At concentrations greater than 0.01 microM, thapsigargin (ThG) dose-dependently caused an increase in cytosolic free Ca2+ concentration ([Ca2+]i) in rat parotid acinar cells, as measured by the fluorescent Ca(2+)-indicator fura-2. In the absence of extracellular Ca2+, a transient increase in [Ca2+]i by ThG was observed, and subsequent addition of carbachol (CCh) did not produce a further [Ca2+]i response, suggesting that ThG released Ca2+ from the CCh-sensitive intracellular Ca2+ pool. Since ThG did not stimulate formation of inositol phosphates, the ThG-induced Ca2+ mobilization is independent of phosphoinositide breakdown. High concentrations (greater than 0.1 microM) of ThG induced amylase release from rat parotide acini, but the effect was very poor as compared with that of CCh or the protein kinase C activator, PMA (phorbol 12-myristate 13-acetate). Combined addition of ThG and PMA modestly potentiated amylase release induced by PMA alone. These results support the view that amylase release by muscarinic stimulation is mediated mainly by activation of protein kinase C rather than a rise in [Ca2+]i, although Ca2+ may modulate the secretory response.
authors.affiliation: Department of Dental Pharmacology, School of Dentistry, Higashi Nippon Gakuen University, Hokkaido, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Tojyo
authors.forename: A
authors.initials: A
authors.lastname: Tanimura
authors.forename: S
authors.initials: S
authors.lastname: Matsui
authors.forename: Y
authors.initials: Y
authors.lastname: Matsumoto
mesh_terms: D000681:Amylases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured / Q000187:drug effects; D003600:Cytosol / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D016257:Fura-2; D007295:Inositol Phosphates / Q000032:analysis; D010306:Parotid Gland / Q000187:drug effects* / Q000378:metabolism; D010703:Phorbol Esters; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000502:physiology*; D013729:Terpenes / Q000494:pharmacology*; D019284:Thapsigargin
chemical_list: D007295:Inositol Phosphates; D010703:Phorbol Esters; D011976:Receptors, Muscarinic; D013729:Terpenes; D019284:Thapsigargin; D002217:Carbachol; D011493:Protein Kinase C; D000681:Amylases; D002118:Calcium; D016257:Fura-2