title: 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.
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Man
authors.initials: M
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Shaoze
authors.initials: S
authors.lastname: Cheng
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Yilin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Ruifeng
authors.initials: R
authors.lastname: Zhao
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China.
authors.forename: Changhua
authors.initials: C
authors.lastname: Sun
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China. ynzhang@yzu.edu.cn.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China. ynzhang@yzu.edu.cn.
authors.forename: Yani
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China. yubcli@yzu.edu.cn.|Province Key Laboratory of Animal Breeding and Molecular Design, Yangzhou, People's Republic of China. yubcli@yzu.edu.cn.
authors.forename: Bichun
authors.initials: B
authors.lastname: Li
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D048430:Cell Shape / Q000187:drug effects; D002645:Chickens / Q000235:genetics / Q000378:metabolism*; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D055785:Gene Knockdown Techniques; D008297:Male; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013094:Spermatozoa / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D014212:Tretinoin / Q000494:pharmacology; D015262:Xenobiotics / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering; D015262:Xenobiotics; D014212:Tretinoin; D019363:Cytochrome P-450 CYP1A1; D019009:Proto-Oncogene Proteins c-kit
keywords: Differentiation; Embryonic stem cells; Metabolism of xenobiotics by cytochrome P450; Primordial germ cells; RNA-Seq; Spermatogonial stem cells