miR-199-3p suppresses cellular migration and viability and promotes progesterone production in goose ovarian follicles before selection through regulating ITGB8 and other ECM-related genes

Abstract

1. The extracellular matrix (ECM) constitutes the basal lamina and the area between follicular cells. Remodelling the ECM is believed to be a key event in follicular development, especially during selection, and plays an important role in cell migration, survival, and steroidogenesis. miR-199-3p is differentially expressed in the goose granulosa layer during follicular selection and is reported to play a primary role in inhibiting cell migration and invasion. Nevertheless, the effect of miR-199-3p on ovarian follicles and its role in follicular cellular migration is not understood.2. In this study, qRT-PCR assays revealed that miR-199-3p was differentially expressed in the granulosa layer from goose ovarian follicles before and after follicular selection. Additionally, miR-199-3p overexpression in cultured granulosa cells (GCs) from goose pre-hierarchical follicles significantly suppressed cell viability and migration. It elevated the concentration of progesterone and the expression of key progesterone production genes. Furthermore, miR-199-3p overexpression in the GCs of goose pre-hierarchical follicles inhibited the expression of ECM-related genes (ITGB8, MMP9 and MMP15) yet promoted the expression of another two ECM-related genes (COL4A1 and LAMA1). Finally, dual-fluorescence reporter experiments on 293T cells established the direct targeting of ECM gene ITGB8 by miR-199-3p.3. In conclusion, miR-199-3p may participate in granulosa cell migration, viability, and steroidogenesis in goose ovarian follicles before selection by modulating ITGB8 and other ECM-related genes.

Keywords: Anser cygnoides; ITGB8; extracellular matrix; granulosa; miR-199-3p; progesterone.