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  • Wiley-Blackwell  (2)
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  • 1
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The hormonal stimulation of phospholipase C and the consequent activation of the Ca2+-phosphatidylinositol cascade in eukaryotic cells is associated with modifications of the [Ca2+] (intracellular Ca2+ concentration) which modulates cellular functions. In this study, these modifications were investigated in primary cultures of human thyroid cells.The mean apparent basal [Ca2+] of human thyrocytes measured using the intracellularly trapped fluorescent indicator Quin-2 was found to be 89°C 16 nM (n = 49). ATP and, to a lesser extent, ADP, but not AMP or adenosine, elicited a concentration-dependent biphasic rise in human thyrocytes [Ca2+] and increased their 45Ca2+ efflux. The first transient phase of the [Ca2+] rise induced by ATP was resistant to extracellular Ca2+ depletion, whereas the second sustained phase was abolished in these conditions. This suggests that although the first phase of this response involves a release of Ca2+ from intracellular stores, the second phase requires extracellular Ca2+ influx. The response of human thyro-cytes to analogs of ATP is compatible with a P2-purinergic effect of ATP on these cells. Bradykinin and TRH affected the human thyrocyte [Ca2+] and 45Ca2+ efflux similarly to ATP. The human thyrocyte [Ca2+] and the 45 Ca2+ efflux were not modified by carbachol, a nonhydrolyzable analog of acetylcholine.The present results suggest the presence of P2-purinergic receptors to ATP and of receptors to TRH and bradykinin on human follicular thyroid cells. They also confirm that the Ca2+-phosphatidylinositol cascade is present in these cells and suggest that this cascade is modulated by ATP, TRH, and bradykinin. As this cascade is involved in the regulation of protein iodination, and therefore of thyroid hormones synthesis, these agents might have an important role in the regulation of the thyroid function.
    Additional Material: 4 Ill.
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  • 2
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Carbachol, through a muscarinic receptor, thyrotropin-releasing hormone (TRH), prostaglandin F2α (PGF2α), bradykinin, and adenosine triphosphate (ATP) in creased the apparent [Ca2+], (intracellular free Ca2+ concentration) of dog thyrocytes in primary culture. The [Ca2+], measured by the Quin-2 technique rose immediately after the addition of the agonists and reached a maximal value after less than 30 seconds. Afterwards, the [Ca2+], declined to a plateau higher than the basal level when the cells were triggered with carbachol. By contrast, in most experiments with PGF2α and in the case of bradykinin, TRH, and ATP, the [Ca2+], returned to the basal value. If the extracellular Ca2+ was chelated by excess of EGTA, the addition of all agents caused a sharp reduced transient rise in the [Ca2+], followed by a decline of the [Ca2+], often below the basal level (especially in the case of carbachol). It is suggested that the first transient phase of these responses is due at least in part to the mobilisation of Ca2+ from intracellular stores whereas the second sustained phase of the response to carbachol mainly originates from an increased Ca2+ influx into the thyrocytes.Carbachol, bradykinin, TRH, PGF2α, and ATP also increased generation of inositol phosphates in dog thyrocytes. This effect was sustained when the cells were triggered with carbachol and was more transient with bradykinin, TRH, PGF2α, or ATP.All these agents and the phorbdester TPA as well as forskolin enhanced to various extent the thyrocyte H2O2 generation. This enhancement was severely reduced in the absence of extracellular Ca2+ and was mimicked by Ca2+ ionophores in the presence of extracellular Ca2+ especially in synergy with protein kinase C activators. These data suggest that the dog thyrocyte H2 O2 generation, the limiting step of the thyroid hormone synthesis, is modulated by carbachol, TRH, PGF2α, bradykinin, and ATP through their action on the Ca2+-phosphatidylinositol cascade.
    Additional Material: 6 Ill.
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