Study On Muscovy Duck Broodiness Using The Techniques Of Proteomics And Metabolomics

Broodiness,one of the main reasons,inhibits the reproduction of Muscovy ducks(Cairina moschata),the mechanism is still unclear.In the study,in order to analyze the molecular level of Muscovy ducks during Nesting and Laying,isotope relative and absolute quantification(iTRAQ)technique and metabolomics and high-throughput technologies were used to analyze the effects of broodiness on global proteins,small molecule metabolites composition,and related signing pathway in Muscovy ducks'ovary.The main results are as follows:1.Proteome analysis of ovary tissue between Nest and LayingUsing the iTRAQ technique,the relative and absolute quantitative analysis of total proteins in ovarian tissue was performed.Each group of 9 ovaries samples,as three biological replicates.The result shows:297472 total spectra were generated.Under the "1%FDR",11708 peptides and 2759 proteins were identified.The Laying as the control group,P<0.05 as the screening criteria,335 differentially expressed proteins were indentified in Nest,with 139 up-regulated and 196 down-regulated.Apovitellenin-1(APOV1),Galanin peptides(GAL),Serum amyloid A protein(SAP),Very low-density lipoprotein receptor Heat shock protein beta-1(HSPB1)were higher up-regulated proteins;Very low-density lipoprotein receptor(VLDLR),Cyclin-dependent kinase 1(CDK1),G-protein(G??),Inositol 1,4,5-trisphosphate receptor type 1(IP3R)were down-regulated.The differentially expressed proteins were analyzed through GO annotated and pathway,we found that Steroid biosynthesis,Calcium signaling pathway,Oocyte meiosis,Steroid hormone biosynthesis,GnRH signaling pathway,PI3K-Akt signaling pathway may be involved in the regulation of muscovy ducks on the broody mechanism.The transcript level of 16 differentially expressed proteins were detected using qRT-PCR.The mRNA expression of 6 genes were inconsistent with iTRAQ analysis,indicating that protein synthesis was regulated by many aspects of reasons;the mRNA expression of 10 genes were almost in accordance with iTRAQ analysis,indicating that iTRAQ technique was feasible and accurate to study the protein composition of ovarian tissue of Muscovy duck.2.Metabolomics analysis of ovary tissue between N and LStudying on the broody of Muscovy ducks by using metabonomics technique,each group of 10 ovaries samples were chosed.The laying as the control group,to study the metabolic map between N and L,we found 10 potential metabolic biomarkers which differ greatly between the two groups.Such as progesterone etc,may be the potential biomarkers for the broodiness.Then,the metabolites were mapped to the KEGG database,mainly enriched in steroid hormone biosynthesis,steroid biosynthesis,amino acid metabolism pathways.Our study revealed the related regulation network of Muscovy duck broodiness by screening the candidate proteins and biomarkers,providing a theoretical basis for the study of Muscovy duck broodiness.