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CYP1A1 based on metabolism of xenobiotics by cytochrome P450 regulates chicken male germ cell differentiation

Abstract

This study aimed to explore the regulatory mechanism of metabolism of xenobiotics by cytochrome P450 during the differentiation process of chicken embryonic stem cells (ESCs) into spermatogonial stem cells (SSCs) and consummate the induction differentiation system of chicken embryonic stem cells (cESCs) into SSCs in vitro. We performed RNA-Seq in highly purified male ESCs, male primordial germ cells (PGCs), and SSCs that are associated with the male germ cell differentiation. Thereinto, the metabolism of xenobiotics by cytochrome P450 was selected and analyzed with Venny among male ESC vs male PGC, male PGC vs SSC, and male ESC vs SSC groups and several candidates differentially expressed genes (DEGs) were excavated. Finally, quantitative real-time PCR (qRT-PCR) detected related DEGs under the condition of retinoic acid (RA) induction in vitro, and the expressions were compared with RNA-Seq. By knocking down CYP1A1, we detected the effect of CYP1A1-mediated metabolism of xenobiotics by cytochrome P450 on male germ cell differentiation by qRT-PCR and immunocytochemistry. Results showed that 17,742 DEGs were found during differentiation of ESCs into SSCs and enriched in 72 differently significant pathways. Thereinto, the metabolism of xenobiotics by cytochrome P450 was involved in the whole differentiation process of ESCs into SSCs and several candidate DEGs: CYP1A1, CYP3A4, CYP2D6, ALDH3B1, and ALDH1A3 were expressed with the same trend with RNA-Seq. Knockdown of CYP1A1 caused male germ cell differentiation under restrictions. Our findings showed that the metabolism of xenobiotics by cytochrome P450 was significantly different during the process of male germ cell differentiation and was persistently activated when we induced cESCs to differentiate into SSCs with RA in vitro, which illustrated that the metabolism of xenobiotics by cytochrome P450 played a crucial role in the differentiation process of ESCs into SSCs.

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Acknowledgments

The authors would like to thank Maozhi Hu, Testing Center, Yangzhou University, Yangzhou, Jiangsu, China, for his interpretation and data analysis. We would like to thank Poultry Institute, Chinese Academy of Agricultural Sciences Experimental Poultry Farm for providing the experimental materials. This work was supported by the National Natural Science Foundation of China (31301959, 31472087). All procedures involving the care and use of animals conformed to US National Institute of Health guidelines (NIH Pub. No. 85-23, revised 1996) and were approved by the laboratory animal management and experimental animal ethics committee of Yangzhou University.

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Correspondence to Yani Zhang or Bichun Li.

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Editor: Tetsuji Okamoto

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Li, D., Wang, M., Cheng, S. et al. CYP1A1 based on metabolism of xenobiotics by cytochrome P450 regulates chicken male germ cell differentiation. In Vitro Cell.Dev.Biol.-Animal 53, 293–303 (2017). https://doi.org/10.1007/s11626-016-0108-z

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Keywords

  • RNA-Seq
  • Embryonic stem cells
  • Primordial germ cells
  • Spermatogonial stem cells
  • Metabolism of xenobiotics by cytochrome P450
  • Differentiation