1,721,065 research outputs found

    Testosterone increases lipolysis and the number of B-adrenoceptors in male rat adipocytes.

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    The influence of androgen status on the regulation of lipolysis and number of beta-adrenoceptors in isolated adipocytes was studied in male rats. Castration resulted in decreased catecholamine-induced as well as forskolin-induced lipolysis. beta-adrenoceptor number, examined by a whole cell cyanopindolol binding assay, was also diminished to a similar extent. Testosterone treatment of castrated rats normalized lipolysis as well as beta-adrenoceptor number. These results demonstrate that testosterone stimulates catecholamine-induced lipolysis in vivo by increasing the number of beta-adrenoceptors as well as the activity of adenylate cyclase, confirming previous in vitro studies performed in adipose precursor cells

    The effects of androgens on the regulation of lipolysis in adipose precursor cells

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    Adipose precursor cells from male rats were exposed in primary culture to testosterone (T) or dihydrotestosterone (DHT), and their effects on the regulation of lipolysis were studied. T, but not DHT, stimulated catecholamine-induced lipolysis in a dose-dependent manner, including physiological concentrations. The effect was equally pronounced with isoproterenol (a pure beta-adrenergic agonist) and norepinephrine (a mixed alpha 2- and beta-adrenergic agonist). The higher lipolytic capacity of catecholamines on T-treated cells was paralleled by a similar increase in the number of beta-adrenoceptors in the cells, without a change in the receptor affinity, suggesting that T induced new synthesis or externalization of beta-adrenoceptors. Both T and DHT stimulated forskolin-induced lipolysis, suggesting an androgen effect at the level of the catalytic subunit of adenylate cyclase. The pertussis toxin-stimulated lipolysis was not influenced by the presence of androgens in the culture medium, and no effect was seen on the antilipolytic effect of insulin. These effects did not disappear in the presence of an aromatase inhibitor, suggesting that the T effects were not mediated by conversion to estrogens. These cells showed specific saturable binding for androgens, with a Kd in the range of androgen concentrations shown to be active. In conclusion, androgens enhance the lipolytic capacity of these cells by increasing the apparent number of beta-adrenoceptors (T only) and the activity of adenylate cyclase (both T and DHT). These changes are not mediated by conversion to estrogens. These effects probably occur via binding to specific androgen receptor

    Up-regulation of androgen receptor binding in male rat fat pad adipose precursor cells exposed to testosterone: Study in a whole cell assay system.

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    Binding of androgens to adipocytes has previously been evaluated using cytosol fractions without taking into account nuclear binding, although the latter is suggested to be close to the physiological site of action. In the present study, performed in differentiated fat pad adipose precursor cells, we describe a simple, reliable and reproducible androgen binding assay in a system with intact cells. Tritiated and unlabeled methyltrienolone (R1881) were used to define specific and unspecific androgen binding. Triamcinolone acetonide was added to prevent the binding of R1881 to other types of receptors. Differentiated adipose precursor cells contain a homogeneous class of high affinity androgen binding sites, and binding is saturable and reversible. Binding apparently occurs at one site, with a Kd in the range of physiological androgen concentration (about 4 nM). Competition studies indicate that the receptor is specific for R1881, testosterone and dihydrotestosterone, which have approximately the same affinity, while progesterone, estradiol and dexamethasone show much lower affinity. Androgen binding was markedly enhanced after cellular exposure to R1881 and testosterone but not dihydrotestosterone, and this increase was dependent on protein synthesis, suggesting the formation of new receptors by these androgens. In conclusion, fully differentiated adipocytes contain a specific, high affinity receptor, the density of which is dependent on androgens

    Estradiol regulation of mRNA expression of stimulatory G-protein alpha subunit in white adipose tissue from female rats.

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    Adipose tissue has been recognized as a major peripheral metabolic target of estrogens. The present study was addressed to examine in female rats whether differences in the adipose tissue mRNA expression of alpha-subunit of stimulatory (Gs) and/or inhibitory (Gi) G-proteins exist between intact and ovariectomized rats, the latter with or without estradiol or testosterone treatment. The fat cell membrane protein amount of Gs and Gi alpha-subunit also was examined. All these parameters were evaluated in parametrial fat tissue samples obtained from 40 female Sprague-Dawley rats. A group of rats (N = 20) was investigated for evaluation of mRNA expression and another group (N = 20) for quantification of the protein amount of Gs and Gi alpha-subunit. Each group was represented by five control rats (sham-operated), five ovariectomized (OVX) rats, five ovariectomized rats treated with estradiol (OVXE) and five ovariectomized rats treated with testosterone (OVXT). Ribonucleic acid extracted from adipose tissue and analyzed by northern blot with G alpha s, G alpha i-3 cRNA probes revealed three major bands with estimated sizes of 1.9, 3.5 and 2.35 kb, respectively. Messenger RNA quantitative analysis, by a solution of hybridization RNAase protection assay on total nucleic acid samples, showed that the amount of G alpha i-1 and G alpha i-2 mRNA was similar within the different groups, whereas the G alpha s mRNA was significantly less abundant (p < 0.01) in OVX and OVXT rats than in control or OVXE rats. No difference in G alpha s mRNA content was found between control and OVXE rat

    Messenger RNA of G-proteins alpha subunit in rat brown adipose tissue.

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    The present study was addressed to quantify the steady-state mRNA levels for the alpha subunit of stimulatory (Gs) and inhibitory (Gi-1 and Gi-2) G-proteins in brown (interscapular) male rat adipose tissue (n = 6 rats). The quantification of specific mRNA, estimated using a solution hybridization RNAse protection assay, showed that the amounts of G alpha i-1, G alpha i-2 and G alpha s mRNA were 0.88 +/- 0.28 amol/microgram DNA, 76 +/- 4 amol/micrograms DNA and 460 +/- 16 amol/micrograms DNA, respectively. When the amounts of G alpha i-1 and G alpha i-2 and G alpha s mRNA in brown adipose tissue were compared with those in epididymal white adipose tissue (obtained from the same rats), G alpha i-1 and G alpha i-2 mRNA levels were very similar between brown and white adipose tissue, whereas the level of G alpha s mRNA was significantly higher in brown than in white fat tissue (P < 0.001). In conclusion, the present study shows the steady-state levels of mRNA for the alpha subunit of Gs, Gi-1 and Gi-2 in rat brown fat and suggests that the quantity of G alpha s mRNA is higher in brown than in white adipose tissue. Further studies are needed to explain the possible physiological importance of these findings

    Testosterone treatment of ovariectomized rats: effects on lipolysis regulation in adipocytes.

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    Alcohol consumption and synthesis of ethyl esters of fatty acids in adipose tissu

    The metabolism of ethyl esters of fatty acids in adipose tissue of rats chronically exposed to ethanol

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    The concentration of ethyl esters of fatty acids as well as the activity of the enzyme synthesizing these esters (fatty acid ethyl ester synthase) were determined in adipose tissue of rats ingesting ethanol (9-16 g/kg body weight/day) for different periods of time. After 10 and 17 weeks of ethanol exposure about 300 nmol of ethyl esters of oleic, palmitic, stearic, and linoleic acids were found per gram adipose tissue. The ethyl esters disappeared after 1 week of abstinence. Closer analyses, using radioactive ethanol, revealed a half-life of the esters of less than 24 hr. The bulk of the esters was found in a membrane preparation of isolated adipocytes. Hormone-sensitive lipase hydrolyzed emulsified ethyl oleate as efficiently as that of trioleoylglycerol, but in mixed ethyl oleate/trioleoyl glycerol particles the hydrolysis of ethyl oleate was slower, suggesting a decreased accessibility. Synthase activity was found in adipose tissue from rats not exposed to ethanol. It doubled after 10 and 17 weeks of ethanol and decreased with a half-life of at least a week after abstinence. It was concluded that ethyl esters of fatty acids are formed in rat adipose tissue as previously shown in other tissues. They seem to be stored mainly in membranous parts of the adipocytes. Synthase activity is induced by ethanol. The elevated activity has a longer half-life, and may be useful as an indicator of alcohol abuse
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