Proteomic Analysis Of Healthy And Atresia Porcine Follicular Granulosa Cells

In mammalian ovaries,99%of follicles undergo atresia at different stages of development.Granulosa cell apoptosis is the initiating factor of follicular atresia after mammal birth.Protein is the executor of life activities.Comparing the granulosa cell protein expression profile of healthy and atretic follicles is of great significance for uncovering the mechanism of mammalian follicular atresia,improving the reproductive potential of excellent female breeding stock and treating some types of infertility.In this study,the protein expression profiles of granulosa cells collected from healthy and atretic porcine follicules of 1?2 mm in diameter were analyzed and compared with tandem mass spectrometry(TMT).Differentially expressed proteins were performed GO enrichment analysis and KEGG signaling pathway analysis,aimed at screening some key proteins that regulate follicular atresia,and laying the foundation for elucidating the mechanism of follicular atresia.Results are as follows:(1)Protein expression profile of healthy follicular granulosa cells(HFGC)and atresia follicular granulosa cells(AFGC)samples were analyzed with TMT,and a total of 6221 proteins were identified.After t-test analysis,499 differentially expressed proteins were screened,of which 327 proteins were significantly up-regulated and 172 proteins were significantly downregulated in the AFGC group.The up-regulated protein with the highest fold is SLC43A2(FC:11.44813),and the down-regulated protein with the highest fold is SMIM1(FC:11.5),which may serve as a granule biomarker for follicular atresia.(2)GO enrichment analysis found that differentially expressed proteins mainly gathered in biological processes such as inflammatory regulation,cell homeostasis,mitochondria transport along microtubules,phagocytosis,cell components such as cytoskeleton,extracellular matrix,and mitochondrial outer membrane,molecular functions such as endopeptidase inhibitor activity,calcium ion binding,GTP binding,GTPase activity,phospholipid binding.(3)KEGG pathway enrichment analysis of differential protein showed AFGC changed in Jak-STAT signaling pathway,estrogen signaling pathway,autophagy signaling pathway,and p53 signaling pathway significantly compared with HFGC.(4)10 differentiated expressed proteins were randomly selected for verifying the reliability of TMT with Western blot technology and results manifested that TMT analysis were true and reliable.In summary,the protein expression profiles of granulosa cells changed significantly when follicle undergoing atresia;SLC43A2 and SMIM1(FC:11.5)may be used as markers of follicular atresia;Jak-STAT signaling pathway,estrogen signaling pathway,autophagy signaling pathway,p53 and other signaling pathways may play an important role in follicular atresia.