mTOR Controls Ovarian Follicle Growth by Regulating Granulosa Cell Proliferation

We have shown that inhibition of mTOR in granulosa cells and ovarian follicles results in compromised granulosa proliferation and reduced follicle growth. Further analysis here using spontaneously immortalized rat granulosa cells has revealed that mTOR pathway activity is enhanced during M-phase of the cell cycle. mTOR specific phosphorylation of p70S6 kinase and 4E-BP, and expression of Raptor are all enhanced during M-phase. The predominant effect of mTOR inhibition by the specific inhibitor Rapamycin (RAP) was a dose-responsive arrest in the G1 cell cycle stage. The fraction of granulosa cells that continued to divide in the presence of RAP exhibited a dose-dependent increase in aberrant mitotic figures known as anaphase bridges. Strikingly, estradiol consistently decreased the incidence of aberrant mitotic figures. In mice treated with RAP, the mitotic index was reduced compared to controls, and a similar increase in aberrant mitotic events was noted. RAP injected during a superovulation regime resulted in a dose-dependent reduction in the numbers of eggs ovulated. Implications for the real-time regulation of follicle growth and dominance, including the consequences of increased numbers of aneuploid granulosa cells, are discussed

Immunolocalization of aquaporin 5 during rat ovarian follicle development and expansion of the preovulatory cumulus oophorus.

Immunofluorescent localization of aquaporin 5 (AQP5) was investigated in rat ovarian follicles during development and preovulatory cumulus oophorus expansion. Ampullary cumuli oophori complexes (COCs) were examined. Analysis revealed that AQP5 immunostaining appeared in preantral follicles and formed a characteristic ring encircling and touching the oolemma. The staining represented most likely AQP5 functioning at the ends of corona radiata cell projections, anchoring on the oocyte surface. However, several hours after the presumptive preovulatory LH surge, when the process of expansion of COCs started, the AQP5 staining appeared also on the cumulus granulosa cells and in the extracellular matrix. In the postovulatory ampullary COCs the fluorescent ring was not observed, which may be the result of the well-established preovulatory withdrawal of projections from the zona pellucida. At that time-point immunofluorescent staining of AQP5 appeared in most oocytes and was also present in the apical membrane of epithelial cells of the oviduct ampulla. The latter observation suggests that after ovulation AQP5 is involved in the transcellular movement of water in the oviduct ampulla and oocytes in rats

Immunolocalization of aquaporin 5 during rat ovarian follicle development and expansion of the preovulatory cumulus oophorus.

Immunofluorescent localization of aquaporin 5 (AQP5) was investigated in rat ovarian follicles during development and preovulatory cumulus oophorus expansion. Ampullary cumuli oophori complexes (COCs) were examined. Analysis revealed that AQP5 immunostaining appeared in preantral follicles and formed a characteristic ring encircling and touching the oolemma. The staining represented most likely AQP5 functioning at the ends of corona radiata cell projections, anchoring on the oocyte surface. However, several hours after the presumptive preovulatory LH surge, when the process of expansion of COCs started, the AQP5 staining appeared also on the cumulus granulosa cells and in the extracellular matrix. In the postovulatory ampullary COCs the fluorescent ring was not observed, which may be the result of the well-established preovulatory withdrawal of projections from the zona pellucida. At that time-point immunofluorescent staining of AQP5 appeared in most oocytes and was also present in the apical membrane of epithelial cells of the oviduct ampulla. The latter observation suggests that after ovulation AQP5 is involved in the transcellular movement of water in the oviduct ampulla and oocytes in rats