Membrane transport of L-triiodthyronine by human red cell ghosts.
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Biomedical subjects
Publications and source records attributed to C Jacquemin.
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The "phospholipid effect" which is the enhanced turnover of the phosphorylinositol group of phosphatidylinositol (PI) occurs in the thyroid of response to thyreostimulin (TSH). The possibility that Ca2+ ions are involved in this stimulation has been investigated with pig thyroid slices. Experiments performed in media without Ca2+ or containing E.G.T.A. (2 mM), indicate that it is not the extracellular Ca2+ which is implied, but rather the intracellular Ca2+. The ionophore A23187 (6.10(-6) M) increases the specific radioactivity of the acid soluble precursors, but has also a specific effect on the PI turnover, which is additive with the effect of a high concentration of TSH (50 mU/ml). Washing and loading of slices with various Ca2+ concentrations show that 0.9 mM restores the TSH phospholipid effect. Verapamil (10(-3) M) and Chlorpromazine (10(-3) M) redirect glycerolipid metabolism by increasing PI and phosphatidic acid (PA) synthesis at the expense of other glycerolipids, as phosphatidylcholine (PC) and phosphatidylethanolamine (PE). These results suggest that the "phospholipid effect" is not a result of Ca2+ entry into the thyroid cells. On the contrary, it seems that this increased turnover of PI in "long term" incubations (3 hr). An additive and acute effect of TSH effect is more pronounced when Ca2+ movements
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Carbenoxolone slightly but significantly decreased the release of FFA from rat epididymal fat pads. The antilipolytic action of carbenoxolone was not blocked by 10(-3)M 3-isobutyl-1-methylxanthine, a potent inhibitor of phosphodiesterase. The findings suggest that carbenoxolone exerts its antilipolytic activity by acting on adenylate cyclase, thereby decreasing cyclic AMP concentrations and the activity of the hormone-sensitive lipase in adipose tissue.
In pig and sheep thyroid, the arachidonate content of phosphatidylinositol (14.3--17.5%) is much higher than in the other phospholipids. In the thyroid, phosphatidylinositol seems to play an important role in the biosynthesis of prostaglandins. In neutral thyroid lipids, the arachidonate content is much higher in diacylglycerols (15.8%) than in monoacylglycerols (2.9%) or triacylglycerols (4.2%). However, the most important pool of esterified arachidonate is triacylglycerols (1030 nmol of arachidonate/g tissue). Arachidonate represents a very small part of total free fatty acids measured in the presence of albumin and indomethacin (0.65% or 16.4 nmol/g tissue). Thyrotropin (50 munits/ml) causes after 1 h a 2-fold increase in the level of free arachidonate (37.3 nmol/g tissue). Pig thyroid slices rapidly take up [14C]arachidonate and incorporate it into neutral lipids and phospholipids. Specific activity of phosphatidylinositol is 3-fold higher than that of phosphatidylcholine after an hour incubation. Specific activity of diacylglycerols is 8-fold higher than that of triacylglycerols. Thyrotropin (50 munits/ml) causes a significant decrease (32--33%) of incorporation of radioactivity in lipids as compared with standard incubation. This result is compatible with the isotopic dilution of the labeled [14C]arachidonate by nonradioactive arachidonate liberated in the incubation medium under thyrotropin action. Slices and homogenates of pig thyroid weakly convert [14C]arachidonate to prostaglandins E2 and F2alpha (1--2%. Thyrotropin (50 munits/ml) always diminishes the conversion of radioactivity to prostaglandins as compared with standard incubation. This result is compatible with the above-mentioned hypothesis.
Lipolysis in rat fat pads was studied by determination of free fatty acid and glycerol production in various experimental conditions (in the absence or presence of glucose, adrenalin and insulin). These results were compared to the accumulation of endogenous prostaglandins E2 and F2alpha during lipolysis. In the absence of glucose the prostaglandin production followed the adrenalin-induced fluctuations in released free fatty acids both in the presence or absence of insulin. In the presence of glucose and insulin, a drop in prostaglandin accumulation was observed whereas free fatty acids production was strongly stimulated. These results suggest that either free fatty acid composition is modified, influencing the activity of prostaglandin synthetase, or that there exists a specific mechanism controlling prostaglandin synthesis.
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The liberation of arachindonate in the thyroid occurs at the expense of two distinct pools of precursors. (1) the phosphatidylinositol through a process Ca2+-dependent and cyclic AMP-independent; and (2) the triglycerides by a cyclic AMP-dependent lipase, in which the involvement of cyclic AMP-dependent protein kinase has not yet been determined. The "PI pool" or "paracyclic AMP pool" is mobilized very rapidly by large doses of TSH but its physiological significance can be discussed. The "triglyceride pool" or "post-cyclic AMP pool" is mobilized more slowly by small doses of TSH and seems not to be implicated in the acute TSH stimulation of adenylate cyclase. The "post-cyclic AMP pool" of prostaglandins would be very important as third messenger or as "long-acting TSH hormone". Some recent works of Boeynaems and Van Sande (16) and Madaoui et al. (17) on the thyroid support this hypothesis, as aspirin or indomethacin inhibits DBc-AMP stimulation of glucose oxydation, iodine organification, or thyroid hormone secretion. On the other hand, in the absence of prostaglandin synthesis, TSH still stimulates the adenylate cyclase, which means that prostaglandins are not obligatory intermediates of hormonal action on cyclic AMP production. In conclusion, these results show a TSH action in the thyroid on the release of fatty acids, precursors of PG's, from their lipidic stores. Nevertheless, a second control step is not excluded in conversion of cyclic endoperoxide to PGE or PGFalpha.
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