Follicle stimulating hormone receptor (FSHR) is a G protein-coupled receptor (GPCR) essential for reproductive function. Localised to the somatic granulosa cells of the ovarian follicle, FSHR is a key driver of antral follicle growth, supporting oocyte developmental competence. To mediate its physiological functions, FSH/FSHR activates Gαs/cAMP/PKA signalling. Recent evidence has suggested additional and alternative signalling mechanisms by which FSH/FSHR can signal. Such mechanisms include the endosomal compartments, which trafficking to were thought
to terminate signalling but have since emerged as a key determinant of signal propagation. Most studies have used heterologous cell systems overexpressing GPCRs to identify the functional roles, with a lack of understanding of the relevance in physiological cell types. This work aimed to determine the relationship between FSHR trafficking and signal activation in primary granulosa cells. We performed kinome-wide activity profiling in primary murine granulosa cells which identified novel signal activation and pathway crosstalk under distinct temporal, spatial and ligand-dependent control. These data established a framework of how FSHR location modulates ligand-dependent signalling signatures and the impact of signal duration in a physiologically relevant cell type, with ramifications for ovarian function. Given the central role of exogenous FSH in assisted reproduction, defining where and when FSHR signals offer a rational basis on which future gonadotrophin preparations might be selected or engineered.