title: Alpha 1A- and alpha 1B-adrenergic receptors mediate the effect of norepinephrine on cytosolic calcium levels in rat PC C13 thyroid cells: thyrotropin modulation of alpha 1B-linked response via a adenosine 3',5'-monophosphate-protein kinase-A-dependent pathway.
abstract: The aim of the present study was to characterize the adrenergic receptors mediating the effects of norepinephrine on PC C13 rat thyroid cells and identify the molecular mechanisms by which TSH regulates the noradrenergic response. We studied TSH regulation of norepinephrine-induced cytosolic calcium increase by means of the fluorescent probe fura-2. In PC C13 cells grown and maintained in a medium containing TSH (PC C13 6H), norepinephrine caused a higher increase in cytosolic calcium than in PC C13 starved from TSH 5 days before the experiments (PC C13 5H). In both group of cells the calcium response to norepinephrine was concentration dependent and reduced by the removal of extracellular calcium ions. Reintroduction of TSH in the culture medium of the PC C13 5H cells induced the recovery of the norepinephrine-stimulated intracellular calcium rise similarly to that in the native PC C13 6H. This effect was complete after a 48-h incubation period and was abolished by the simultaneous treatment of the cells with the protein synthesis inhibitor cycloheximide, suggesting that TSH may stimulate the synthesis of alpha 1-adrenergic receptors in PC C13 cells. Because in these cells we found that TSH increased cAMP levels as well as inositol phosphate production, we tested whether the activation of a protein kinase-A and/or protein kinase-C was involved in TSH regulation of the adrenergic response. We found that the treatment of PC C13 5H cells with forskolin restored the effect of norepinephrine on the calcium level, and that KT5720, an inhibitor of the protein kinase-A, was able to prevent the recovery of the noradrenergic response induced by the readdition of TSH to the culture medium of PC C13 5H. Conversely, treatment of PC C13 5H cells with the protein kinase-C activator phorbol 12-myristate 13-acetate was ineffective. Norepinephrine also stimulated inositol phosphate production in PC C13 6H and, to a lesser extent, in PC C13 5H, but it did not affect the cAMP levels in the two groups of cells. To characterize alpha 1-adrenergic receptor subtypes mediating the effects of norepinephrine in PC C13 cells, we used antagonists of alpha 1A and alpha 1B receptors (WB4101 and chlorethylclonidine respectively).(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Dipartimento di Scienza delle Comunicazioni Umane, Facoltà di Medicina e Chirurgia, Università di Napoli Federico II, Italy.
authors.forename: O
authors.initials: O
authors.lastname: Meucci
authors.forename: A
authors.initials: A
authors.lastname: Scorziello
authors.forename: A
authors.initials: A
authors.lastname: Avallone
authors.forename: C
authors.initials: C
authors.lastname: Ventra
authors.forename: M
authors.initials: M
authors.lastname: Grimaldi
authors.forename: M T
authors.initials: MT
authors.lastname: Berlingieri
authors.forename: A
authors.initials: A
authors.lastname: Fusco
authors.forename: G
authors.initials: G
authors.lastname: Schettini
mesh_terms: D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D007295:Inositol Phosphates / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D009638:Norepinephrine / Q000494:pharmacology*; D051381:Rats; D011942:Receptors, Adrenergic, alpha / Q000502:physiology*; D013961:Thyroid Gland / Q000166:cytology / Q000378:metabolism*; D013972:Thyrotropin / Q000494:pharmacology
chemical_list: D000317:Adrenergic alpha-Antagonists; D002121:Calcium Channel Blockers; D007295:Inositol Phosphates; D011942:Receptors, Adrenergic, alpha; D013972:Thyrotropin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D002118:Calcium; D009638:Norepinephrine