| Hens reproductive performance is a dynamics process,controlled by several factors.It is reported that the mechanism of ovary development differs between mammal and poultry,especially in hormone regulation.Aromatase(encoded by CYP19A1),is a crucial enzyme reposed to the conversion from androgen to estrogen.In this study,we performed NGS on chicken small yellow follicles to get insight in chicken follicles transcriptomics.Bioinformatics were conducted to characterize the gene expression pattern of theca cells layers(TCs)and granulosa cells(GCs).Subsequently,transcriptomics comparison were conducted between cow and chicken follicles,and potential transcription factors of CYP19A1 were filtered according to gene expression pattern within 2 species.The potential transcription factors were verified by dual-luciferase reporter system finally.The main results were as followed:(1)According to CYP19A1 expression pattern within chicken ovary,aromatase showed high expression in TCs rather than GCs,and peaked at small yellow follicles stage;(2)Transcriptomics profile within TCs and GCs of chicken small yellow follicles revealed 179 TC-specific expressed genes,196 TC-biased expressed genes,18 GC-specific expressed genes and 39 GC-biased expressed genes;(3)Weighted gene co-expression network analysis(WGCNA)were conducted in each dataset respectively,and a high similarity between chicken_SYF and cow_D dataset were revealed according to the analysis.Besides,two modules in chicken_SYF were filtered which turned out to have high correlation to CYP19A1 expression;(4)A further analysis was conducted to chicken_SYF and cow_D datesets,and ESR2 were selected as the potential transcription factor of CYP19A1 based on the different gene expression pattern between chicken and cow;(5)Confirmatory experiments were conducted by dual-luciferase reporter system at cell level to confirm the inhibiting effect of ESR2 on CYP19A1 promoter activity. |
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