Nitric oxide promotes differentiation of rat white preadipocytes in culture.

The putative role of nitric oxide (NO) in modulating adipogenesis was investigated in cultured preadipocytes derived from rat white adipose tissue. The NO releasing reagent, hydroxylamine (HA), and nitric oxide synthase (NOS) substrate L-arginine (Arg) had no influence on cell replication. However, both HA and Arg exhibited significant induction on differentiation, as evidenced by increased lipoprotein lipase (LPL) and glycerol-3-phosphate dehydrogenase (GPDH) activities, as well as accelerated triacylglycerol (TG) accumulation.

Development of insulin resistance in the JCR:LA-cp rat: role of triacylglycerols and effects of MEDICA 16.

The JCR:LA-cp rat develops an extreme obese/insulin-resistant syndrome such that by 12 weeks of age, there is no longer any insulin-mediated glucose turnover. At 4 weeks of age, obese and lean rats have essentially identical basal and insulin-mediated glucose uptake in skeletal muscle. By 8 weeks of age, however, the obese rats no longer exhibit such intake.

Long-term regulation of leptin expression is correlated with adipocyte number in obese rats.

Objective: To investigate long-term regulation of leptin expression in adipose tissues of obese JCR:LA-corpulent rats, which have been shown to overexpress leptin.

Design: Manipulation of adipose tissue growth in obese rats by dietary restriction.

Interventions: Weanling female obese rats were maintained on 1 of 3 diets until 8 months old.

Arachidonic acid metabolites of the lipoxygenase as well as the cyclooxygenase pathway may be involved in regulating preadipocyte differentiation.

Conditions that trigger preadipocyte differentiation in vivo have yet to be elucidated. To investigate the role of endogenous arachidonic acid (AA) metabolites on adipose tissue growth, rat preadipocytes in primary culture were induced to differentiate using medium conditioned by isolated mature adipocytes (ACM). Differentiation was determined by assay of glycerol-3-phosphate dehydrogenase (GPDH).

A novel method for studying preadipocyte differentiation in vitro.

In vitro differentiation of rat preadipocytes has typically been induced in medium supplemented with pharmacological concentrations of hormonal mixtures. These conditions probably do not reflect the milieu within adipose tissue in vivo. We have developed a new method for inducing differentiation of preadipocytes using culture medium which has been conditioned by isolated adipocytes (ACM).

Overexpression of the obese gene in the genetically obese JCR:LA-corpulent rat.

Expression of the obese (ob) gene in JCR:LA-cp rats was examined. A 360 bp fragment of the conserved region of the gene was obtained by RT-PCR using total RNA isolated from adipose tissues of Sprague-Dawley (SD), JCR:LA-cp obese and lean rats. The three gene fragments were sequenced and shown to be identical.

Exogenous triacylglycerol inhibits insulin-stimulated glucose transport in L6 muscle cells in vitro.

This study tested the hypothesis using cultured L6 myocytes that insulin resistance in muscle may be the consequence of triacylglycerol accretion in the tissue itself. Exposure of L6 myocytes to triacylglycerol for 4 hours resulted in significant transfer of lipid into the cells compared to control cells treated for only 5 min. Insulin-stimulated 2-deoxyglucose uptake in L6 myocytes was reduced when the cells were preloaded with triacylglycerol.

Regulation of new fat cell formation in rats: the role of dietary fats.

Factors that stimulate formation of new adipocytes during development of obesity are yet to be identified. We examined whether diet acts directly on preadipocytes to stimulate replication and differentiation or indirectly by interacting with adipocytes to release or modify local growth factors. Male Sprague-Dawley rats were fed chow or diets high in starch (HST), saturated (HFS) or polyunsaturated (HFP) fats until 5-7 months of age.

Fatty acid synthase and adipsin mRNA levels in obese and lean JCR:LA-cp rats: effect of diet.

In Sprague-Dawley rats, fatty acid synthase (FAS) activity is suppressed by dietary fat. To test the hypothesis that a defect in regulation of de novo fatty acid synthesis exists in massive obesity, we investigated the effect of diet on FAS mRNA levels in genetically obese JCR:LA-corpulent (cp) rats. We also determined levels of mRNA encoding adipsin, a fat cell-derived protein possibly associated with lipid metabolism.

Hepatic and adipose tissue lipogenic enzyme mRNA levels are suppressed by high fat diets in the rat.

Small changes in lipogenic enzyme activity induced by dietary fats of different composition may, over the long term, have significant impact on the development of obesity. We have investigated the effect of high fat diets (45% of calories as fat) on abundance of mRNA encoding fatty acid synthetase (FAS) and glycerophosphate dehydrogenase (GPDH) in male Sprague-Dawley rats. When caloric intake was equal, the relative amount of hepatic FAS mRNA was greater in rats fed a saturated compared to a polyunsaturated fat diet.

Paradoxically slow preadipocyte replication and differentiation in corpulent rats.

We have investigated the in vitro rate of replication and differentiation of preadipocytes derived from lean (+/+) and obese (cp/cp) male JCR:LA-corpulent (cp) rats in an attempt to identify mechanisms that regulate adipose tissue growth. Cp/cp rats were twofold heavier than age-matched lean rats by 9-10 mo. Cp/cp-derived preadipocytes demonstrated an inherently slower rate of replication than +/+ preadipocytes (population doubling time: cp/cp 52.

Influence of paracrine factors on preadipocyte replication and differentiation.

Regional adipose tissue growth may be modulated by paracrine factors that influence preadipocyte replication and/or differentiation. To investigate this hypothesis, we have studied the effects of culture media conditioned by adipose microvascular endothelial cells, preadipocytes, or mature fat cells, on rat preadipocyte replication and differentiation in vitro. Endothelial cell-conditioned medium (ECCM) stimulated preadipocyte replication while medium conditioned by mature fat or preadipocytes had little effect.

Induction of preadipocyte differentiation by mature fat cells in the rat.

In this study we investigated the influence of mature adipocytes, derived from rat adipose tissue, on the replication and differentiation of preadipocytes in primary culture. Mature fat did not inhibit preadipocyte replication within the 6-d period studied. Preadipocyte differentiation, as assessed by both cytoplasmic lipid accretion and an increase in glycerophosphate dehydrogenase (GPDH) activity, was significantly stimulated by the presence of mature fat tissue or isolated adipocytes.

Endogenous and exogenous cholecystokinin may reduce food intake by different mechanisms.

To determine whether exogenous cholecystokinin (CCK) and endogenous CCK evoke different gastrointestinal motor responses, we investigated the motility induced by CCK by use of standard manometric methods. Injection (ip) of 500 ng/kg CCK caused immediate profound gastric inhibition and duodenal phasic excitation that did not resemble the postprandial pattern. Similar profound gastric inhibition has been associated with nausea.

Proglumide, a cholecystokinin antagonist, increases gastric emptying in rats.

Injection of cholecystokinin (CCK) reduces food intake and delays gastric emptying. We have previously shown that endogenous CCK also reduces food intake. This may be achieved by a delay in gastric emptying.

The effect of vagotomy on the increase in food intake induced by the cholecystokinin antagonist, proglumide.

Cholecystokinin, secreted when ingested food enters the duodenum, may act as a satiety factor. Injection of proglumide, a specific antagonist of cholecystokinin, induced an increase in food intake. The satiety effect of administered cholecystokinin is abolished by bilateral subdiaphragmatic vagotomy.

The cholecystokinin antagonist, proglumide, increases food intake in the rat.

Cholecystokinin, secreted in response to ingested food entering the duodenum, may play a role in limiting food intake. Inhibition of cholecystokinin should therefore induce an increase in food intake. Proglumide, a specific antagonist of cholecystokinin was used to block the satiety effect of a food preload in rats.