Study Of Interaction Mechanisms Between Goose Follicle Granulosa And Theca Cells

The follicular development of poultry is closely related to their egg production.Granulosa cell(GC)and theca cell(TC)are important components of the follicles,and their interaction is of great significance to the growth and development of follicles.However,the study on the interaction between GC and TC in poultry and its mechanism is very limited.Therefore,in this study,the geese used as the experiment object to explore the development characteristics of granulosa and theca tissue during its follicular development.An in vitro co-culture model of GC and TC was constructed in each follicular development stage,the effects of interaction on the physiological functions of GC and TC were examined,and the mechanism of interaction between GC and TC was analyzed by using transcriptome and proteome techniques.The main results are as follows:1.The results of follicular histology showed that the morphology of granulosa layer and theca layer changed significantly with the development of follicle.In the follicles less than 2 mm,the theca layer was not completely formed;and the morphology and thickness of the granulosa layer and the theca layer tissue in follicles 2-10 mm,F4-F2 and F1 were significantly different.2.Established the in vitro co-culture model of pre-hierarchical(2-10 mm),hierarchical(F4-F2)and F1 follicle GC and TC by transwell system.It was found that in the co-culture model,the cell activity,lipid metabolism and DNL,steroidogenesis,proliferation and apoptosis pathways of GC and TC changed significantly in all the stages.3.For co-culture group GC(C-GC)vs.mono-culture group GC(M-GC),the number of DEG was 1338 at 48 h,359 at 96 h,and the number of DEP was 455.And for co-culture group TC(C-TC)vs.mono-culture group TC(M-TC),the number of DEG was 423 at 48 h,539 at 96 h,and the number of DEP was 74.It is suggested that C-GC had more differential genes and proteins than C-TC.4.DEG and DEP of C-GC are mainly enriched in c GMP-PKG,TGF? and PI3 K signaling pathways related to proliferation and apoptosis;phospholipid D and PPAR signaling pathways related to lipid metabolism.DEG and DEP of C-TC are mainly enriched in Wnt and PI3 K signaling pathways related to proliferation and apoptosis;glycerolipid metabolism and AMPK,PPAR signaling pathway related to lipid metabolism.These pathways are closely related to the changes of physiological functions of C-GC and C-TC.5.The trend and time point of gene/protein changes of BMP4,VIP and CXCL14 in C-TC were highly consistent with that of BMPR2-TGF?,VIPR2-c AMP and CXCR4-PI3 K in C-GC,while the VEGFA in C-GC were highly consistent with that of VEGFR2-PI3K/RAP1 in C-TC.In summary,this study analyzed the interaction mechanism of GC and TC,and revealed the important roles of BMP4-BMPR2,VIP-VIPR2,CXCL14-CXCR4 and VEGFA-VEGFR2 pathways in the process of GC and TC interaction on their proliferation,apoptosis and steroidogenesis,which laid a theoretical foundation for further understanding the mechanism of goose follicular development.