Single-cell transcriptomic analysis reveals regulative mechanisms of follicular selection and atresia in chicken granulosa cells.

Eggs are an important food source for people. Follicle selection and atresia are the two directions of pre-hierarchical follicles that affect egg production in chickens. Granulosa cells (GCs), the vital somatic cells in follicles, determine the fate of follicles. In this study, single-cell RNA sequencing was performed on the GC layers from five follicular stages (small white follicles, atretic small white follicles, small yellow follicles, atretic small yellow follicles, and F6) to map the cellular differentiation trajectories and explore the follicle fate-determining genes. The results showed that GCs were genetically heterogeneous and could be divided into four subtypes, and the presence of GCs-Ⅲ with a steroid-producing capacity in unselected small follicles is a novel finding that differs from conventional wisdom. In addition, degenerated GCs were annotated for the first time, and GC degeneration was found to be significantly related to lipid metabolism disorders. Many candidate switch genes had been marked out, among which the overexpression of transforming growth factor-beta 2 (TGFB2) and insulin like growth factor binding protein 5 (IGFBP5) could inhibit the proliferation and differentiation of GCs and induce their degeneration. This study provided new insights into the regulatory mechanisms of follicle selection and atresia, which have significant value for improving egg production and prolonging the laying period of laying hens.