Comparative Analysis of Transcriptome of Pigeon Follicles at Early Stage of Laying Interval Reveals Genes Related to Follicular Development
High throughput sequencing was performed on granulosa cell(GC)layer at early stage of laying interval(LI)to screen key genes involved in pigeon follicle development and selection.Eighty pairs of 12-month-old White King pigeons,with similar laying patterns and body weights were selected in this study.Three female pigeons were selected at first day(LI1)and third day(LI3)of LI,respectively,the GC layer of F1 and SF1 follicles were collected,followed by tran-scriptome analysis using RNA-seq.Differentially expressed genes(DEGs)among 4 groups:L1F1/L1SF1 group,L3F1/L3SF1 group,L1F1/L3F1 group,and L1SF1/L3SF1 group were ana-lyzed.Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Ge-nomes(KEGG)pathway enrichment analysis of DEGs were performed,protein-protein interac-tion(PPI)network analysis using the STRING database and visualization using Cytoscape soft-ware were conducted to selected key genes.Five DEGs from each group were randomly selected for RT-qPCR to verify the reliability of transcriptome.The 77,2 736,5 698 and 3 864 DEGs were obtained in the 4 groups,respectively.GO analysis results showed that DEGs in all groups were mainly enriched in cellular processes,cellular anatomical entity and binding.KEGG analysis result showed that DEGs in L1F1/L1SF1 and L3F1/L3SF1 groups were enriched in the steroid hormone biosynthesis and ovarian steroidogenesis signaling pathways.DEGs in L1F1/L3F1 and L1SF1/L3SF1 groups were enriched in the glycosaminoglycan degradation,synaptic vesicle cycle and oxidative phosphorylation signaling pathways.In addition,DEGs closely associated with fol-licle development and selection were screened by PPI.Five DEGs were randomly selected from each group for RT-qPCR verification,and the expression trends of these DEGs were consistent with the sequencing results.RNA-seq technology was applied to identify key genes associated with pigeon follicle development and selection,which including EEF2,DDX5,PLK2,IGF2R,LHCGR,HSD3B1,CYP19A1 and StAR.These results will provide the theoretical basis for further exploring the molecular mechanisms of follicle development and selection in pigeon.
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