Figure 1

Expression of adiponectin system (mRNA and proteins) in bovine ovary and embryo. (A) RT-PCR analysis of the mRNAs for adiponectin, AdipoR1 and AdipoR2 in small (SF) and large (LF) follicles, corpus luteum (CL), whole ovary (Ov), cumulus cells from immature (Cum Im) and 24-h IVM (Cum IVM) COC, immature (Oo Im) and 24-h IVM (Oo IVM) oocytes, granulosa cells (GC) and adipose tissue (AT). (B) Detection of adiponectin (30 kDa), AdipoR1 (42 kDa), AdipoR2 (44 kDa) and APPL1 (82 kDa) by immunoblotting in LF, SF, OV, CL, AT, cumulus cells (Cum, from 40 24-h IVM COC per lane), fresh (fGC) and 48-h cultured (cGC) granulosa cells from small follicles (< 6 mm), follicular fluid (FF), oocytes (Oo, n = 100 per lane; except for APPL1, n = 20 per lane) and embryo (Em, in vitro day-8 blastocyst, n = 55 per lane). Rat adipose tissue (rAT) was used as positive control for the presence of adiponectin and APPL1, as already tested in our laboratory with the present antibodies. Mouse muscle (mM) and mouse liver (mL) were used as positive controls for the presence of AdipoR1 and AdipoR2 respectively, because these two antibodies were described to cross react with mouse. Vinculin protein was used as a loading control (n = 3).