title: Stimulation of generation of inositol phosphates by carbamoylcholine and its inhibition by phorbol esters and iodide in dog thyroid cells.
abstract: The action of carbamoylcholine (Cchol), NaF and other agonists on the generation of inositol phosphates (IPs) was studied in dog thyroid slices prelabelled with myo-[2-3H]inositol. The stimulation by Cchol (0.1 microM-0.1 mM) of IPs accumulation through activation of a muscarinic receptor [Graff, Mockel, Laurent, Erneux & Dumont (1987) FEBS Lett. 210, 204-210] was pertussis- and cholera-toxin insensitive. Ins(1,4,5)P3, Ins(1,3,4)P3 and InsP4 were generated. NaF (5-20 mM) also increased IPs generation (Graff et al., 1987); this effect was potentiated by AlCl3 (10 microM) and unaffected by pertussis toxin. Although phorbol dibutyrate (5 microM) abolished the cholinergic stimulation of IPs generation (Graff et al., 1987), it did not affect the fluoride-induced response. Cchol and NaF did not require extracellular Ca2+ to exert their effect, and neither KCl-induced membrane depolarization nor ionophore A23187 (10 microM) had any influence on basal IPs levels, or on cholinergic stimulation. However, more stringent Ca2+ depletion with EGTA (0.1 or 1 mM) decreased basal IPs levels as well as the amplitude of the stimulation by Cchol without abolishing it. Dibutyryl cyclic AMP, forskolin, cholera toxin and prostaglandin E1 had no effect on basal IPs levels and did not decrease the response to Cchol. Iodide (4 or 40 microM) also strongly decreased the cholinergic action on IPs, this inhibition being relieved by methimazole (1 mM). Our data suggest that Cchol activates a phospholipase C hydrolysing PtdIns(4,5)P2 in the dog thyroid cell in a cyclic AMP-independent manner. This activation requires no extracellular Ca2+ and depends on a GTP-binding protein insensitive to both cholera toxin and requires no extracellular Ca2+ and depends on a GTP-binding protein insensitive to both cholera toxin and pertussis toxin. The data are consistent with a rapid metabolism of Ins(1,4,5)P3 to Ins(1,3,4)P3 via the Ins(1,4,5)P3 3-kinase pathway, followed by dephosphorylation by a 5-phosphomonoesterase. Indeed, a Ca2+-sensitive InsP3 3-kinase activity was demonstrated in tissue homogenate. Stimulation of protein kinase C and an organified form of iodine inhibit the Cchol-induced IPs generation. The negative feedback of activated protein kinase C could be exerted at the level of the receptor or of the receptor-G-protein interaction.
authors.affiliation: Department of Endocrinology, Hôpital Erasme, Brussels, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Laurent
authors.forename: J
authors.initials: J
authors.lastname: Mockel
authors.forename: K
authors.initials: K
authors.lastname: Takazawa
authors.forename: C
authors.initials: C
authors.lastname: Erneux
authors.forename: J E
authors.initials: JE
authors.lastname: Dumont
mesh_terms: D000535:Aluminum / Q000494:pharmacology; D000077410:Aluminum Chloride; D017607:Aluminum Compounds; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology / Q000502:physiology; D002217:Carbachol / Q000494:pharmacology*; D002712:Chlorides / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology; D004285:Dogs; D066298:In Vitro Techniques; D007295:Inositol Phosphates / Q000378:metabolism*; D007454:Iodides / Q000494:pharmacology*; D010703:Phorbol Esters / Q000494:pharmacology*; D010770:Phosphotransferases / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D011976:Receptors, Muscarinic / Q000502:physiology; D012969:Sodium Fluoride / Q000494:pharmacology; D013961:Thyroid Gland / Q000378:metabolism*
chemical_list: D017607:Aluminum Compounds; D002712:Chlorides; D007295:Inositol Phosphates; D007454:Iodides; D010703:Phorbol Esters; D011976:Receptors, Muscarinic; D000001:Calcimycin; D000077410:Aluminum Chloride; D002217:Carbachol; D012969:Sodium Fluoride; D000535:Aluminum; D000242:Cyclic AMP; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase; D002118:Calcium