Resolving Candidate Genes for Chicken Ovarian Transplantation through RNA-seq and WGCNA
Qin, QLiu, RDing, XLi, ZZhang, YYi, XZhao, Y
This study aimed to identify candidate genes regulating ovarian development following ovarian tissue transplantation through transcriptome sequencing (RNA-seq) and weighted gene co-expression network analysis (WGCNA). Ovarian tissues were collected from 10 thirty-day-old donor chickens, and each ovary was divided equally into three parts of similar size, and transplanted in situ into two-day-old recipient chickens. Samples were collected on days 0 (untransplanted), 6, and 12 after transplantation. RNA was extracted from ovarian samples for construction of a transcriptome library, and then sequenced on the IlluminaHiSeqTM2000 sequencing platform. The sequencing results were evaluated to determine differentially expressed genes (DEGs) through GO function and the KEGG pathway analyses, Gene Set Enrichment Analysis (GSEA), WGCNA, and PPI networks. Some candidate genes were further validated through real-time fluorescence quantitative PCR (RT-qPCR). RNA-seq analysis identified 2242 up-regulated and 3095 down-regulated DEGs at 6 days after ovarian transplantation, and 2181 up-regulated and 2129 down-regulated DEGs were identified at 12 days after transplantation. Enrichment analysis showed that the genes were enriched in multiple inflammatory pathways, pathways involved in signal transduction and cellular processes. Through the WGCNA, five modules were constructed, from which 4 and 5 candidate genes were mined at 6 and 12 days after ovarian transplantation, respectively. Transcription factor prediction showed that GTF3A was the most important transcription factor. The results of RT-qPCR verification confirmed that the expression profile of 8 candidate genes was consistent with that of the sequencing results. In summary, this study presents the candidate genes involved in ovarian vascular remodeling and ovarian proliferation after ovarian transplantation.(AU)
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