LAUSR

Abstract Androgens mediate a number of processes in mammalian and teleost ovaries in a follicle-stage dependent manner, including follicle growth, survival, and apoptosis. We recently reported that the membrane androgen receptor ZIP9 mediates apoptosis in Atlantic croaker granulosa/theca (G/T) cells from mature ovarian follicles, but the effects of androgens on early stage G/T cells in this model remains unknown. Here we show that testosterone mediates pro- and anti-apoptotic responses in a follicle stage-dependent manner in croaker ovarian follicle cells. Testosterone treatment decreased the incidence of apoptosis in G/T cells from early stage follicles (diameter <300 µm) but increased apoptosis in G/T cells from late stage follicles (diameter >400 µm). Small interfering RNA targeting ZIP9, but not the nuclear androgen receptor, blocked the anti-apoptotic response, indicating ZIP9 mediates anti-apoptotic in addition to pro-apoptotic responses. Testosterone treatment of early stage G/T cells resulted in opposite signaling outcomes from those previously characterized for the ZIP9-mediated apoptotic response including decreased cAMP and intracellular free zinc levels, and downregulation of pro-apoptotic member mRNA expression. While ZIP9-mediated apoptosis involves activation of a stimulatory G protein (Gs), activators of Gs signaling antagonized the anti-apoptotic response. Proximity ligation and G protein activation assays indicated that in G/T cells from early stage follicles ZIP9 is in close proximity and activates an inhibitory G protein, while in G/T cells from late stage follicles ZIP9 is in close proximity and activates Gs. This study demonstrates that ZIP9 mediates opposite survival responses of croaker G/T cells by activating different G proteins in a follicle stage-dependent manner. Summary Sentence The membrane androgen receptor ZIP9 mediates pro- and anti-apoptotic responses in teleost ovarian follicle cells by activating stimulatory and inhibitory G proteins, respectively, in a follicle stage-dependent manner