DNA methylation-mediated transcription factors regulate Piwil1 expression during chicken spermatogenesis

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  • QIU Lingling
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • XU Lu
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • CHANG Guobin
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • GUO Qixin
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • LIU Xiangping
    Poultry Institute, Chinese Academy of Agricultural Sciences, Jiangsu 225003, China
  • BI Yulin
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • ZHANG Yu
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • WANG Hongzhi
    Poultry Institute, Chinese Academy of Agricultural Sciences, Jiangsu 225003, China
  • WANG Kehua
    Poultry Institute, Chinese Academy of Agricultural Sciences, Jiangsu 225003, China
  • LU Wei
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • REN Lichen
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • ZHU Pengfei
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • WU Yun
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • ZHANG Yang
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • XU Qi
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China
  • CHEN Guohong
    College of Animal Science & Technology, Yangzhou University, Jiangsu 225009, China

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  • DNA methylation-mediated transcription factors regulate <i>Piwil1</i> expression during chicken spermatogenesis

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<p> The P-element induced wimpy testis (Piwi) protein family is responsible for initiating spermatogenesis and maintaining the integrity of germ cells and stem cells, but little is known regarding its transcriptional regulation in poultry. Here, we characterized the methylation status of the Piwil1 promoter in five different spermatogenic cell lines using direct bisulfite pyrosequencing and determined that methylation correlates negatively with germ cell type-specific expression patterns of piwil1. We demonstrated that methylation of the −148 CpG site, which is the predicted binding site for the transcription factors TCF3 and NRF1, was differentially methylated in different spermatogenic cells. This site was completely methylated in PGCs (primordial germ cells), but was unmethylated in round spermatids. A similar result was obtained in the region from +121 to +139 CpG sites of the Piwil1 promoter CpG island, which was predicted to contain SOX2 binding sites. In addition, demethylation assays further demonstrated that DNA methylation indeed regulates Piwil1 expression during chicken spermatogenesis. Combined with transcription factor binding site prediction, we speculate that methylation influences the recruitment of corresponding transcription factors. Collectively, we show the negative correlation between promoter methylation and piwil1 expression and that the spatiotemporal expression of chicken Piwil1 from the PGC stage to the round spermatid stage is influenced by methylation-mediated transcription factor regulation.</p>

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