The Function Of PPAR Gamma In Lipid Metabolism Of Goose Ovary Granulosa Cells

The egg production of geese is influenced by factors such as feeding environment,method,hormone level and follicular development,among which follicular development is the most direct factor determining egg production.During follicular development,the proliferation and apoptosis of granule cells and the changes of follicular steroid hormones were accompanied by the increase of follicles.Lipid metabolism from scratch by fatty acids,fatty acid synthesis and decomposition of beta oxidation,triglycerides,in which PPARgamma play an important role.This experiment was going to clone the PPARgamma CDS region and promoter sequences,and to analysis the expression of PPARgamma and other related genes by using real time fluorescence quantitative techniques.At the same time,by interfering and over-expressing the PPARgamma in cultured granulosa cells,the key genes’ expression related to lipid metabolism,cholesterol synthesis pathway were analyzed.The main results are as follows:(1)The CDS region and the promoter sequence of goose PPARgamma gene were cloned and sequenced,which were 1428 bp and 2795 bp respectively.The similarity of geese PPARgamma is the highest with other birds,all above 98%,which conforms to the genetic evolution of species.The PPARgamma promoter has 4 core promoter regions with multiple transcription factor binding sites.(2)In the pre-grade follicles,the expression of PPARgamma in the membrane layer is higher than that of the simultaneous granular layer.The expression of PPARgamma in the F5 stage is the highest,and the expression of PPARgamma in the membrane layer is higher than that of the granular layer at the F1 stage.The expression of PPARgamma in the granular layer of the follicles in the F5 to F2 stage is higher than that of the simultaneous membrane.(3)After interfering the gene expression of PPARgama,the expression of FAS gene in granulosa cells increased but not significant(P>0.05),and the expression of ACC gene increased significantly(P<0.05).The expression level of ApoB and CPT-1 was reduced but not significant(P>0.05).The expression of SREBP1,DGAT1 and DGAT2 genes increasedsignificantly(P<0.05).The expression of CCND2 gene was significantly decreased(P<0.05),and the expression of STAR and SREBP2 gene was increased but not significant(P>0.05).(4)After over-expressing the gene of PPARgama,the expression of ApoB gene in the granulosa cells increased but not significant(P>0.05),the expression of CPT1 gene increased significantly(P<0.05),the FAS gene’s expression decreased but not significant(P>0.05),the ACC gene’s expression decreased significantly(P<0.05),the DGAT2 gene’s expression decreased significantly(P<0.01)and the SREBP2 gene’s expression decreased but not significant(P>0.05).The expression of CCND2 gene increased significantly(P<0.05),STAR gene’s expression decreased significantly(P<0.01),and the BCL2 gene’s expression decreased significantly(P<0.01).