Fetal baboon sex-specific outcomes in adipocyte differentiation at 0.9 gestation in response to moderate maternal nutrient reduction.

OBJECTIVE: To investigate in vitro adipocyte differentiation in baboon fetuses in response to reduced maternal nutrition. DESIGN: Cross-sectional comparison of adipocyte differentiation in normally grown fetuses and fetuses of pregnant baboons fed 70% of the control global diet from 30 days of pregnancy to term. SUBJECTS: The subjects comprised control (CTR) fetuses (five female and five male) of mothers fed ad libitum and fetuses of mothers fed 70% of the global diet consumed by CTR (maternal nutrient reduction (MNR), five female and five male fetuses). The expression of genes/proteins involved in adipogenesis (PPARγ, FABP4 and adiponectin) and brown adipose tissue development (UCP1, TBX15 and COXIV) were determined in in vitro-differentiated stromal-vascular cultures from subcutaneous abdominal, subcutaneous femoral and omental adipose tissue depots. Adipocyte number per area (mm(2)) was determined histologically to assist in the evaluation of adipocyte size. RESULTS: Maternal suboptimal nutrition suppressed growth of male but not female fetuses and led to adipocyte hypertrophy accompanied by increased markers of white- and, particularly, brown-type adipogenesis in male but not female fetuses. CONCLUSION: Adipose tissue responses to fetal nonhuman primate undernutrition are sexually dimorphic. While female fetuses adapt adequately, the male ones enhance pathways involved in white and brown adipose tissue development but are unable to compensate for a delayed development of adipose tissue associated with intrauterine growth restriction. These differences need to be considered when assessing developmental programming of adiposity in response to suboptimal maternal nutrition.

Commonality versus specificity among adiposity traits in normal-weight and moderately overweight adults.

BACKGROUND: Many adiposity traits have been related to health complications and premature death. These adiposity traits are intercorrelated but their underlying structure has not been extensively investigated. We report on the degree of commonality and specificity among multiple adiposity traits in normal-weight and moderately overweight adult males and females (mean body mass index (BMI)=22.9 kg m(-2), s.d.=2.4). METHODS: A total of 75 healthy participants were assessed for a panel of adiposity traits including leg, arm, trunk, total fat masses and visceral adipose tissue (VAT) derived from dual energy X-ray absorptiometry (DXA), hepatic and muscle lipids from proton magnetic resonance spectroscopy, fat cell volume from an abdominal subcutaneous adipose tissue biopsy (n=36) and conventional anthropometry (BMI and waist girth). Spearman's correlations were calculated and were subjected to factor analysis. RESULTS: Arm, leg, trunk and total fat masses correlated positively (r=0.78-0.95) with each other. VAT correlated weakly with fat mass indicators (r=0.24-0.31). Intrahepatic lipids (IHL) correlated weakly with all fat mass traits (r=0.09-0.34), whereas correlations between DXA depots and intramyocellular lipids (IMCL) were inconsequential. The four DXA fat mass measures, VAT, IHL and IMCL depots segregated as four independent factors that accounted for 96% of the overall adiposity variance. BMI and waist girth were moderately correlated with the arm, leg, trunk and total fat and weakly with VAT, IHL and IMCL. CONCLUSION: Adiposity traits share a substantial degree of commonality, but there is considerable specificity across the adiposity variance space. For instance, VAT, IHL and IMCL are typically poorly correlated with each other and are poorly to weakly associated with the other adiposity traits. The same is true for BMI and waist girth, commonly used anthropometric indicators of adiposity. These results do not support the view that it will be possible to identify adequate anthropometric indicators of visceral, hepatic and muscle lipid content in normal-weight and moderately overweight individuals.

Committed subcutaneous preadipocytes are reduced in human obesity.

AIMS/HYPOTHESIS: The aim of this study was to test whether the availability of committed preadipocytes in abdominal and femoral subcutaneous adipose tissue varies with obesity and body fat distribution. METHODS: Body composition, fat cell size, committed preadipocytes and macrophages were measured in subcutaneous abdominal and femoral adipose depots of 17 lean, 16 upper-body-obese (UBO) and 13 lower-body-obese (LBO) women. Preadipocytes and macrophages were identified by simultaneous staining with the respective markers aP2 and CD68. In a subset of samples we measured preadipocyte proliferation, differentiation and susceptibility to apoptosis. RESULTS: Abdominal adipocytes were smaller in lean than in obese women. Committed preadipocytes represented a greater fraction of stromovascular cells in lean than in obese women but were similar between UBO and LBO women (abdomen: approximately 30 +/- 3 vs approximately 17 +/- 2%; thigh: approximately 30 +/- 3 vs approximately 17 +/- 2%). Preliminary data suggested that preadipocyte kinetics were similar in LBO and lean women, whereas preadipocytes of UBO women differentiated less and were more susceptible to apoptotic stimuli. The fraction of stromovascular cells that were macrophages was greater in both depots in obese women (UBO and LBO) than in normal-weight women, but the difference was not statistically significant. CONCLUSIONS/INTERPRETATION: The proportion of subcutaneous adipose tissue stromovascular cells that are committed preadipocytes is reduced with obesity. This could be due to greater recruitment of preadipocytes to adipogenesis or greater preadipocyte apoptosis, depending upon the obesity phenotype. These data are consistent with the concept that body fat distribution may be regulated partly through differences in adipogenesis.

Tumor necrosis factor-alpha binding in porcine primary stromal-vascular cell cultures.

The binding characteristics of tumor necrosis factor-alpha receptors (TNFRs) in primary stromal-vascular cultures from fat tissue of 7-d-old pigs were analyzed. Cells were plated and maintained in 10% fetal bovine serum from day 0 to day 3 and then switched to serum-free medium from day 3 to day 6 to induce lipid filling. On days 3 and 6 of culture, some of the cells were lysed for ligand and immunoblotting and the remainder subjected to competitive and inhibitory-binding assays. Media from day 6 of culture were subjected to ligand and immunoblotting. Competitive binding analysis showed one-site bindings, with IC50s in the nanomolar and Kds in the picomolar ranges, that were not significantly different at both time-points of measurement. However, the Bmax decreased significantly with differentiation. Preincubation with antibody against TNF receptor type 1 (TNFR1) or TNF receptor type 2 reduced the specific binding by 95 and 15%, respectively, suggesting a dominating role of TNFR1 in 125I-labeled TNFalpha (125I-TNFalpha) binding. This was further supported by ligand blotting of cell lysates. Ligand and immunoblotting of cell lysates indicated that TNFalpha utilizes both types of surface receptors and their isoforms which were not modified during differentiation. Ligand blotting of media revealed soluble receptors with high Mr implying the formation of multimers. Immunoblotting suggested the presence of both types of TNFRs, but a greater abundance of soluble TNFR1. Also, it indicated the additional formation of smaller oligomers from both types of soluble receptors suggesting higher affinity of larger multimers for 125I-TNFalpha.

Priming with magnesium-deficient media inhibits preadipocyte differentiation via potential upregulation of tumor necrosis factor-alpha.

The effect of priming stromal-vascular cells in primary cultures with magnesium-deficient (MgD) media on preadipocyte differentiation was studied. Cultures were derived from dorsal subcutaneous fat tissue of young pigs and maintained 3 d in serum-free control or MgD Dulbecco's modified Eagle's medium, 3 d in 10% fetal bovine serum and dexamethasone, and 6 d in insulin. At d 12 of culture, immunocytochemical and glycerophosphate dehydrogenase assays indicated depressed adipocyte differentiation in the MgD groups. Cultures were enriched for preadipocytes up to 50% of total cells. On the third day of treatment with control and MgD medium, total cell lysates were isolated and 50 microg of them were run on two-dimensional gel electrophoresis. The separated proteins from both treatment groups showed similar patterns. However, spots of proteins with predicted molecular weight in the range of 25.8-37.4 kDa and pI of 5.39-5.85 were sixfold denser from the MgD 10 groups than from the controls. These proteins migrate similarly to tumor necrosis factor-alpha (TNF-alpha). The amount of TNF-alpha in cell lysates from the MgD group was about 2.35 times greater than controls determined by TNF-alpha-ELISA. It is likely that proteins upregulated by MgD medium are TNF-alpha isoforms.

Enhancing effect of troglitazone on porcine adipocyte differentiation in primary culture: a comparison with dexamethasone.

OBJECTIVE: This study compares the effects of the thiazolidinedione, troglitazone (TGZ), dexamethasone (DEX), and DEX plus TGZ on preadipocyte differentiation and the expression of CCAAT/enhancer-binding protein-alpha (C/ EBPalpha) and peroxisome proliferator-activated receptor-gamma (PPARgamma). RESEARCH METHODS AND PROCEDURES: Adipose tissue was obtained from postnatal pigs to isolate stromal-vascular cells. First, we applied 1, 5, or 10 microM TGZ and 10% fetal bovine serum for 3 days and counted the number of recruited preadipocytes. Next, we used either 10 microM TGZ, 80 nM DEX, or DEX plus TGZ with 10% fetal bovine serum for 3 days and then switched to serum-free medium with insulin for 6 days. On day 3 of culture, we counted preadipocytes, and on days 3 and 6 of culture, we performed immunostaining and Western blot analysis to determine the expression of C/EBPalpha and PPARgamma proteins. On day 9 of culture, we stained for lipids with oil red-O and measured the activity of glycerol phosphate dehydrogenase. RESULTS: DEX and TGZ equally enhanced recruitment of preadipocytes and late differentiation, but these effects were not additive with DEX plus TGZ treatment. However, TGZ and DEX had a differential effect on morphogenesis; DEX-treated adipocytes were larger and organized in loose clusters, whereas TGZ-treated cells were smaller and formed compact clusters. Both agents increased C/EBPalpha expression but in a temporally distinct manner. DEX was a better inducer than TGZ, and its effect was early and temporary. However, treatment with either TGZ or DEX did not change PPARgamma protein expression as evaluated by a Western blotting, but immunocytochemistry showed a tendency for increased numbers of PPARgamma positive cells. DISCUSSION: TGZ and DEX equally enhance early and late adipocyte differentiation, possibly by using some common pathways for preadipocyte recruitment. The differential effect on morphogenesis implies a potential differential effect on the expression of extracellular matrix components. C/EBPalpha may be the critical transcription factor involved in TGZ- and DEX-induced adipogenesis.