Effect of gonadotropin treatment on size, number, and cell proliferation of antral follicles in cows

To determine the effects of gonadotropins on the size, number, and cell proliferation of antral ovarian follicles, cows received FSH-P or vehicle beginning on Day 2 after estrus, and ovaries were collected 6, 12, 24, or 48 hr after the initiation of FSH-P treatment or 24 or 48 hr after the initiation of vehicle treatment. Ovaries also were collected from untreated cows on Day 2 after estrus (pretreatment). Before fixation, all visible antral follicles were counted and their surface diameters were recorded. Proliferating cells were immunolocalized in fixed follicles by using a specific primary antibody against proliferating cell nuclear antigen (PCNA), and the labeling index (LI; percentage of cells staining positively for PCNA) was determined for granulosa and thecal cells. After 48 hr of treatment, FSH-P-treated cows had fewer (P < 0.01) small antral follicles and more medium and large antral follicles (P < 0.01 and P < 0.05, respectively) compared with vehicle-treated cows. Granulosa cell LI was negatively correlated (P < 0.05) with follicular diameter for vehicle-treated but not for FSH-P-treated cows. Analysis of covariance using follicular diameter as a covariate to adjust to a common diameter indicated that granulosa cell LI was greater (P < 0.05) at 24 and 48 hr in FSH-P-treated than in vehicle-treated cows; conversely, thecal cell LI was greater (P < 0.01) at 48 hr in FSH-P-treated compared with vehicle-treated cows but did not differ at 24 hr. Across all groups, the LI of cells located within the antral half of the granulosa cell layer was greater (P < 0.01) than that of cells located within the basal half. In conclusion, the stimulation of follicular development by exogenous gonadotropins increased or maintained the proliferation of granulosa and thecal cells concomitant with continued follicular growth. Therefore, enhanced follicular cell proliferation may be an important mechanism by which FSH-P superinduces the growth of antral follicles in cows.