Figure 7

PGD2 signaling controls the granulosa cell proliferation. (A), The length of estrous cycles in five WT and five HQL-79-treated adult mice were assessed in vaginal smears collected every day for 16 consecutive days. Results of the five animals were averaged and were expressed as means +/- SE (colums and bars), * P value = 0.0097. (B), Proliferation of granulosa cells of antral follicles was assessed using immunofluorescence with mitosis marker phosphohistone H3 (phosphoH3) antibody (in red) on cryosections of wild type (-HQL-79) or HQL-79 (+HQL-79) treated ovaries; granulosa cells were identified by anti-Müllerian hormone (AMH) antibody (in green) and nuclei were labeled by the Hoescht Dye (HST) (in blue). Numbers of phospho-H3-positive cells were determined on ten independent fields of three different ovaries for each condition and are represented on the graphs (C). * represents significant increased number of mitotic cells in HQL-79 treated compared to that in untreated ovaries. (D), CyclinD2 and p27 expression levels in five wild type and five HQL-79-treated ovaries were quantified by real time RT-PCR and were normalized to Gapdh expression. Values are the result of averaged experiments (done in triplicate) on the five independent ovaries. * represents the significant decrease of p27 expression in HQL-79 compared to that in untreated ovaries (P-value < 0.025). (E), The follicular content of HQL-79 treated ovaries (at their proestrous stage) were compared to that of WT ovaries by labeling sections with the Hoescht dye. CL: corpora lutea, * growing follicles.