Figure 1

p2y2r, p2y4r , and p2y6r expression in mouse theca cells. (A) RNA from cultured theca cells (1), whole ovary (2), heart (3), and brain (4) was reverse transcribed and amplified by PCR for p2Y2, p2Y4, p2Y6, cyp11A, cyp 17A, star, fshr, and β-actin using specific oligonucleotides to detect transcript expression. Controls in lanes (5) and (6) correspond to a PCR reaction of a theca cell RNA sample without reverse transcriptase and to the reaction mix without a cDNA template, respectively. Amplicon length is indicated in base pairs (bp). Each amplification assay was repeated in 3-5 independent cultures. (B) CREB phosphorylation evaluated by Western blot from TIC cultures in basal conditions or stimulated for 10 min by either 2 IU hCG or 1 ng/ml FSH. The 43-kDa band was analyzed by densitometry, and the result (mean ± S.E.M) of three determinations from independent cultures was plotted (*p < 0.01). PARP protein was used as gel loading control. (C) P2Y2 and P2Y6 receptors detected by Western blot (WB), from freshly isolated TIC homogenates directly (P2Y2) or after immunoprecipitation (IP) (P2Y6).