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Biphasic effects of glucagon and cyclic AMP on the synthesis and secretion of lipids by rat hepatocytes.

ABSTRACT: Cultured rat hepatocytes were preincubated with glucagon or a cyclic AMP analogue for up to 24 h and lipid synthesis and secretion were determined during the next 2 h. Glucagon or cyclic AMP did not change the incorporation of choline or glycerol into phosphatidylcholine, or choline into sphingomyelin, in the cells after 0-12 h of preincubation. After 12 h these incorporations were increased. Incorporations into hepatic lysophosphatidylcholine were decreased after preincubation with glucagon or cyclic AMP for 0-12 h, but by 24 h they increased. There was no change in the lysophosphatidylcholine in the medium after preincubation with glucagon or cyclic AMP for up to 6 h, but increases occurred after preincubation from 12 to 24 h. The secretion of triacylglycerol was decreased after preincubation for 0-1 h, but it returned to control values after 4 h. After preincubation for 18-24 h the incorporation of glycerol into secreted triacylglycerol was increased. The results are discussed in relation to the control of lipid metabolism in starvation and diabetes.

SUBMITTER: Hermier D

PROVIDER: S-EPMC1151503 | biostudies-other | 1991 Nov

REPOSITORIES: biostudies-other

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Project description:The effects of hepatocyte volume on the secretion of triacylglycerol were studied in order to test the suggestion that increases in the portal concentrations of osmolyte amino acids and metal ions during the prandial/early-absorptive phase may be involved in mediating the acute changes in glycerolipid metabolism observed in vivo [Zammit (1995) Biochem Soc. Trans. 23, 506-511]. Incubation of isolated rat hepatocytes with hypo-osmotic medium or in the presence of glutamine (in the presence or absence of leucine), conditions which gave an increase in cell water content of between 8 and 27%, resulted in a decrease in the rate of [14C]triacylglycerol (TAG) secretion when [14C]palmitate was used as substrate. The inhibition was proportional to the increase in cell water content. At low exogenous palmitate concentration (0.05 mM), the inhibition of [14C]TAG secretion was accompanied by a marked shift in the incorporation of label from TAG to phospholipid. In the presence of 0.5 mM palmitate this effect was attenuated, and in the presence of 1 mM palmitate it was abolished. Increased cell volume associated with incubation of hepatocytes with glutamine (in the presence or absence of leucine) also resulted in a decrease in the fraction of newly labelled TAG that was secreted into the medium. Decreased cell volume, achieved by incubation of hepatocytes with hyperosmotic medium (sufficient to decrease cell water content by approx. 9%) decreased overall [14C]TAG secretion, but did not affect the amount of label that was incorporated into phospholipid as a fraction of that incorporated into total glycerolipids. Cell shrinkage, however, diminished the fraction of newly labelled [14C]TAG that was secreted. When intracellular TAG was prelabelled with [3H]glycerol, it was found that cell shrinkage markedly inhibited (preformed) [3H]TAG secretion, whereas cell swelling did not affect this route of TAG secretion. The data are discussed in terms of the possible action of changes in cell hydration at the different loci at which hepatocyte TAG secretion is controlled, with reference to previous observations that both insulin and glucagon are able to inhibit TAG secretion in cultured rat hepatocytes and HepG2 cells.

Project description:The effects of dopamine on pituitary prolactin secretion and pituitary cyclic AMP accumulation were studied by using anterior pituitary glands from adult female rats, incubated in vitro. During 2h incubations, significant inhibition of prolactin secretion was achieved at concentrations between 1 and 10nm-dopamine. However, 0.1-1mum-dopamine was required before a significant decrease in pituitary cyclic AMP content was observed. In the presence of 1mum-dopamine, pituitary cyclic AMP content decreased rapidly to reach about 75% of the control value within 20min and there was no further decrease for at least 2h. Incubation with the phosphodiesterase inhibitors theophylline (8mm) or isobutylmethylxanthine (2mm) increased pituitary cyclic AMP concentrations 3- and 6-fold respectively. Dopamine (1mum) had no effect on the cyclic AMP accumulation measured in the presence of theophylline, but inhibited the isobutylmethylxanthine-induced increase by 50%. The dopamine inhibition of prolactin secretion was not affected by either inhibitor. Two derivatives of cyclic AMP (dibutyryl cyclic AMP and 8-bromo cyclic AMP) were unable to block the dopamine (1mum) inhibition of prolactin secretion, although 8-bromo cyclic AMP (2mm) significantly stimulated prolactin secretion and both compounds increased somatotropin (growth hormone) release. Cholera toxin (3mug/ml for 4h) increased pituitary cyclic AMP concentrations 4-5-fold, but had no effect on prolactin secretion. The inhibition of prolactin secretion by dopamine was unaffected by cholera toxin, despite the fact that dopamine had no effect on the raised pituitary cyclic AMP concentration caused by this factor. Dopamine had no significant effect on either basal or stimulated somatotropin secretion under any of the conditions tested. We conclude that the inhibitory effects of dopamine on prolactin secretion are probably not mediated by lowering of cyclic AMP concentration, although modulation of the concentration of this nucleotide in some other circumstances may alter the secretion of the hormone.

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Biphasic effects of glucagon and cyclic AMP on the synthesis and secretion of lipids by rat hepatocytes.

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