Effect of Estradiol on Granulosa Cells of Goose (Anser cygnoides domestica) Grade Follicles
SU Hang, LAN Gang, HU Shen-Qiang, OUYANG Qing-Yuan, GAN Xiang, CHEN Xi, WANG Ji-Wen*
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
Abstract：Estradiol (E2) plays an important role in regulating the growth and development of granulosa cells.To explore the effect of estradiol (E2) at different concentrations on Goose (Anser cygnoides domestica) hierarchical follicular granulosa cells (HFGCs) and its molecular mechanism, in this study, the grade granulosa cells of 'Tianfu' geese were randomly divided into experimental group (E5, E7, E9) and control group (NC), the granulosa cells of experimental group were treated with different concentrations of estradiol (1×10-5, 1×10-7 and 1×10-9 mol/L). The samples were collected for MTT assay, oil staining assay and transcriptome sequencing analysis. MTT results showed that the cell activity of the experimental group was significantly higher than that of the control group (P<0.05); Oil red staining results showed that with the decrease of E2 concentration, the lipid deposition of granulosa cells decreased, and the difference was significant when E2 concentration was 1×10-9 mol/L (P<0.05); By transcriptome sequencing analysis, E5, E7, E9 were compared with NC under the condition of fold change (FC)>1.2 or <0.83 and P<0.05, 267, 79 and 162 differentially expressed genes were screened, respectively; KEGG enrichment analysis showed that the differentially expressed genes of E5, E7, E9 and NC were significantly enriched in sphingolipid metabolism, insulin signaling and Wnt signaling, ABC transporter and other pathways, respectively. 3-ketodihydrosphingosine reductase (KDSR), ceramide synthase 6 (CERS6) and ceramide synthase 4 (CERS4) in sphingolipid metabolism pathway, mitogen-activated protein kinase 8 (MAPK8), inositol polyphosphate phosphatase like 1 (INPPL) and sclerostin (SOST) in insulin and Wnt signaling pathway were involved in regulating granulocyte activity; Galactose-3-O-sulfotransferase 1 (GAL3ST1), sphingosine-1-phosphate lyase 1 (SGPL1), ATP binding cassette subfamily A member 2 (ABCA2) and ATP binding cassette subfamily A member 5 (ABCA5) in sphingolipid metabolism and ABC transporter pathway were involved in regulating granulosa cells lipid deposition when E2 concentration was 1×10-9 mol/L. The results showed that exogenous E2 treatment could improve the activity of goose granulosa cells and inhibit lipid deposition by affecting sphingolipid metabolism, insulin signaling, Wnt signaling and the expression levels of related genes in ABC transporter pathway. The results lay a foundation for exploring the molecular mechanism of estradiol regulating the growth and development of goose granulosa cells.
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