IS THERE A SPATIAL GRADIENT OF CELL CYCLE REGULATION IN THE DEVELOPING OVARIAN FOLLICLE?
Barrett, Susan1,2, Albertini, David1, 1 Department of Anatomy and Cellular Biology, Boston, MA2 Program in Cellular, Molecular and Developmental Biology, Boston, MA ABSTRACT- The proliferative potential of the somatic granulosa cells appears to be modulated by both paracrine/oocyte derived factors (e.g. Kit ligand, GDF-9) and systemic gonadotropins such as follicle-stimulating hormone (FSH). We have analyzed cell cycle status of mouse granulosa cells (GCs) with respect to stage of follicle development and proximity to the oocyte. GCs were obtained by follicle puncture of ovaries from 14-day-old unprimed prepubertal or 21-day-old eCG primed animals. Mural or cumulus GCs were plated for 4d or 3d, respectively, in serum containing medium and then fixed and processed for immunofluorescence microscopy. Cultures were analyzed for mitotic index (Hoechst 333258), G1/S cell cycle progression (anti-PCNA) or terminal post mitotic differentiation status (TUJ1) and the percentages of cells in each
