Regulation of leptin by agouti.

Dominant mutations at the mouse Agouti locus lead to ectopic expression of the Agouti gene and exhibit diabetes, obesity, and yellow coat color. Obese yellow mice are hyperinsulinemic and hyperleptinemic, and we hypothesized that Agouti directly induces leptin secretion. Accordingly, we used transgenic mice expressing agouti in adipocytes (under the control of aP2 promoter, aP212) to examine changes in leptin levels. Agouti expression in adipose tissue did not significantly alter food intake, weight gain, fat pad weight, or insulinemia; however, the transgenic mice were hyperglycemic. We demonstrated that plasma leptin levels are approximately twofold higher in aP212 transgenic mice compared with their respective controls, whereas ubiquitous expression of agouti (under the control of beta-actin promoter, BAP20) led to a sixfold increase in leptin. Insulin treatment of aP212 mice increased adipocyte leptin content without affecting plasma leptin levels. These findings were further confirmed in vitro in 3T3-L1 adipocytes treated with recombinant Agouti protein and/or insulin. Agouti but not insulin significantly increased leptin secretion, indicating that insulin enhances leptin synthesis but not secretion while Agouti increases both leptin synthesis and secretion. This increased leptin synthesis and secretion was due to increased leptin mRNA levels by Agouti. Interestingly, agouti regulation of leptin was not mediated by melanocortin receptor 4, previously implicated in agouti regulation of food intake. These results suggest that increased leptin secretion by agouti may serve to limit agouti-induced obesity, independent of melanocortin receptor antagonism, and indicate that interaction between obesity genes may play a key role in obesity.

The agouti gene product stimulates pancreatic [beta]-cell Ca2+ signaling and insulin release.

Ubiquitous expression of the mouse agouti gene results in obesity and hyperinsulinemia. Human agouti is expressed in adipose tissue, and we found recombinant agouti protein to stimulate lipogenesis in adipocytes in a Ca(2+)-dependent fashion. However, adipocyte-specific agouti transgenic mice only became obese in the presence of hyperinsulinemia. Because intracellular Ca(2+) concentration ([Ca(2+)](i)) is a primary signal for insulin release, and we have shown agouti protein to increase [Ca(2+)](i) in several cell types, we examined the effects of agouti on [Ca(2+)](i) and insulin release. We demonstrated the expression of agouti in human pancreas and generated recombinant agouti to study its effects on Ca(2+) signaling and insulin release. Agouti (100 nM) stimulated Ca(2+) influx, [Ca(2+)](i) increase, and a marked stimulation of insulin release in two beta-cell lines (RIN-5F and HIT-T15; P < 0. 05). Agouti exerted comparable effects in isolated human pancreatic islets and beta-cells, with a 5-fold increase in Ca(2+) influx (P < 0.001) and a 2.2-fold increase in insulin release (P < 0.01). These data suggest a potential role for agouti in the development of hyperinsulinemia in humans.

Body composition in Chinese subjects: comparison with data from North America.

A multiple tracer dilution method measuring total body water, extracellular water, plasma volume, and red cell volume was used to study body composition. Healthy Chinese subjects were compared to a group of Chinese patients with various newly diagnosed gastrointestinal disorders scheduled for elective operation. Compositional data obtained from these groups showed no significant differences in compartmentalization, suggesting absence of major changes during the early stages of certain gastrointestinal diseases. In addition, healthy Chinese subjects were compared to a group of healthy Americans previously evaluated by similar dilutional methods. In males, body weight (kg) differed considerably (Chinese = 62.1 +/- 2.0 versus American = 72.5 +/- 4.1, p less than 0.02) and could be explained by alterations in body fat (Chinese = 12.4 +/- 1.3 versus American = 19.2 +/- 2.2, p less than 0.02) and extracellular water (Chinese = 14.4 +/- 0.5 versus American = 16.8 +/- 0.8, p less than 0.02). Functional tissue ("body cell mass") was similar in the 2 groups which suggested that Chinese have comparable body composition without an additional fat burden. The multiple tracer technique using deuterium oxide, sodium bromide, Evans dye, and Cr51 for body compositional assessment is accurate but expensive and laborious. Therefore, equally precise but more economical bedside methods are needed for routine compositional analysis.