In keeping with these observations, an additional experiment showed that the age at first ovulation, assessed by the detection of cornified cells (first estrus) in vaginal smears followed by two consecutive days of leukocytes,(a condition highly correlated with ovulation) was delayed several days in LV-EED-injected rats (age at first ovulation: LV-GFP= 33 ± 0.31, n=5 vs LV-EED= 37.8 ±0.6, n=6; t=−16.12, p<0.001, Student t Test), and estrous cyclicity was disrupted (Fig. 8a,b). Examination of the ovaries at 50 days of age showed that LV-EED injected animals had some corpora lutea indicating that they had ovulated, but also exhibited an excess of antral follicles that had not reach the periovulatory stage (Fig. 8c). In contrast, LV-GFP-injected rats had an abundance of corpora lutea indicative of repeated ovulations (Fig. 8c). In a third experiment, the LV-GFP and LV-EED constructs were delivered to the ARC of 22-day-old rats and after all animals in the LV-GFP injected group showed three complete estrous cycles, all the animals were exposed to a fertile male for five days. We observed that rats in which the LV-EED construct
