In rat primary pituitary cells, recombinant adiponectin regulates GnRH receptor expression. In mouse L T2 gonadotrope cells and in rat pituitary cells, recombinant adiponectin inhibits LH secretion. Moreover, adiponectin and AdipoR2 are expressed selleck chemicals llc in rat and human placenta and rat placental adiponectin mRNA increases Inhibitors,Modulators,Libraries in abundance during preg nancy whereas AdipoR2 has the opposite Inhibitors,Modulators,Libraries pattern. Adiponectin and its receptors are also present in Inhibitors,Modulators,Libraries ovary of various species including pig, chicken and rat. Ledoux et al. reported that recombinant adi ponectin has a direct effect on gene expression in porcine follicular cells associated with ovarian follicle remodel ling. In recent work, our group showed that adi ponectin is mainly expressed in rat theca interstitial cells and could exert paracrine effects on the steroidogenesis of granulosa cells.
The expression of the adiponectin system in insulin target tissues in human and animal diabetic models has been studied. However, the expression Inhibitors,Modulators,Libraries of the adiponec tin system in reproductive tissues in diabetic animals has not been explored. We report an investigation of ovarian steroid production and the protein levels of key factors involved in steroido genesis in two conditions in primary rat granulosa cells in the presence of high glucose concentrations. and in strep tozotocin treated female rats. In these conditions, we also assayed adiponectin receptor proteins in ovarian cells. Methods Hormones and reagents Purified ovine FSH 20 used for culture treatment was a gift from NIDDK, National Hormone Pituitary Program, Bethesda, MD, USA.
Recombinant human insulin like growth fac tor I was from Sigma. Recom binant human adiponectin produced in the NSO mammalian cell system was obtained from R D. Glucose was from VWR. Antibodies Rabbit polyclonal antibodies to adiponectin receptor Inhibitors,Modulators,Libraries 2 and adiponectin receptor 1 were from Phoenix Pharmaceuticals Inc.Rabbit polyclonal antibodies to phospho ERK1 2, and phospho AMPK alpha Thr172 were pur chased from New England Biolabs Inc. Rab bit polyclonal antibodies to AMPK 1 were obtained from Upstate Biotechnology Inc. Rab bit polyclonal antibodies to ERK2 were purchased from Santa Cruz Biotechnology. Mouse monoclonal antibody to vinculin was obtained from Sigma. Rabbit polyclonal antibodies against p450scc, StAR and 3 HSD were generously pro vided by Dr. Dale Buchanan Hales and Dr.
Van Luu The, respectively. Mouse monoclonal antibody against p450 aromatase was pur chased from Serotec. All antibodies were used at 1 1000 dilution for western things blotting. Animals and experimental procedures All procedures were approved by the Agricultural Agency and the Scientific Research Agency and conducted in accordance with the guidelines for Care and Use of Agri cultural Animals in Agricultural Research and Teaching. Eight week old female Wistar rats were purchased from Janvier Laboratories.