The expression of peroxisome proliferator-activated receptor gamma in pig fetal tissue and primary stromal-vascular cultures.

Abstract

This study was designed to determine when peroxisome proliferator-activated receptor gamma (PPARgamma) is expressed in developing fetal adipose tissue and stromal-vascular adipose precursor cells derived from adipose tissue. In addition we examined developing tissue for CCAAT/enhancer-binding protein beta (C/EBPbeta) expression to see if it was correlated with PPARgamma expression. Pituitary function and hormones involved with differentiation (dexamethasone and retinoic acid) were also tested for their effects on PPARgamma expression to determine if hormones known to affect differentiation also effect PPARgamma expression in vivo and in cell culture. Developing subcutaneous adipose tissues from the dorsal region of the fetal pig were collected at different gestation times and assayed using Western blot analysis to determine levels of PPARgamma and C/EBPbeta. Hypophysectomy was performed on 75-day pig fetuses and tissue samples were then taken at 105 days for Western blot analysis. Adipose tissue was also taken from postnatal pigs to isolate stromal-vascular (S-V) cells. These adipose precursor cells were grown in culture and samples were taken for Western blot analysis to determine expression levels of PPARgamma. Our results indicate that PPARgamma is expressed as early as 50 days of fetal development in adipose tissue and continues through 105 days. Expression of PPARgamma was found to be significantly enhanced in adipose tissue from hypophysectomized fetuses at 105 days of fetal development (p<0.05). C/EBPbeta was not found in 50- or 75-day fetal tissues and was found only at low levels in 105-day tissues. C/EBPbeta was not found in hypophysectomized (hypoxed) 105-day tissue where PPARgamma was elevated. S-V cells freshly isolated from adipose tissue of 5- to 7-day postnatal pigs showed the expression of PPARgamma1. When S-V cells were cultured, both PPARgamma1 and 2 were expressed after the first day and continued as cells differentiated. High concentrations of retinoic acid decreased PPARgamma expression in early S-V cultures (p<0.05). Our data indicate that PPARgamma is expressed in fetal adipose tissue very early before distinct fat cells are observed and can be expressed without the expression of C/EBPbeta. The increase in PPARgamma expression after hypophysectomy may explain the increase in fat cell size under these conditions. Adipose precursor cells (S-V cells) from 5- to 7-day postnatal pigs also express PPARgamma in the tissue before being induced to differentiate in culture. Thus S-V cells from newborn pig adipose tissue are probably more advanced in development than the 3T3-L1 cell model. S-V cells may be in a state where PPARgamma and C/EBPalpha are expressed but new signals or vascularization are needed before cells are fully committed and lipid filling begins.