Growth differentiation factor 9 (gdf9) and bone morphogenetic protein 15 (bmp15) are potential intraovarian regulators of steroidogenesis in Japanese flounder (Paralichthys olivaceus)

Abstract

Members of transforming growth factor-β (TGF-β) superfamily are vital regulators during the development of fish ovary. However, its intraovarian functions in teleost are still unclear. As members of the TGF-β superfamily, gdf9 and bmp15 are necessary for follicle formation and granulosa cell proliferation. Here in Japanese flounder, quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization (ISH) analysis showed that gdf9 and bmp15 were mainly expressed in oogonia and oocytes, whereas weakly expressed in non-ovarian tissues. Overexpression of single gdf9 and the co-overexpression with bmp15 could up-regulate the expression of most steroidogenic genes, while the overexpression of single bmp15 could down-regulate the expression of most steroidogenic genes. These findings demonstrate that single gdf9 and the combination with bmp15 may act as "activator", while single bmp15 may act as "inhibitor" in the process of steroidogenesis in flounder. This was also verified in negative feedback regulation of gdf9 and bmp15 during hormone treatment. High concentration of human chorionic gonadotropin (hCG) could down-regulate gdf9 and up-regulate bmp15, which were beneficial for the homeostasis of hCG hormone. Besides, knockdown of either gdf9 or bmp15 could significantly down-regulate most steroidogenic genes. This indicated that heterodimer of GDF9:BMP15 might be the most bioactive ligand in gonad development of flounder. Taken together, our study provided a novel recognition that gdf9 and bmp15 could regulate steroidogenesis in teleost through mechanism different from that in mammals.

Keywords: Japanese flounder; Steroidogenesis; bmp15; gdf9.