Epigenetic modification cooperates with Zeb1 transcription factor to regulate Bmp4 to promote chicken PGCs formation

Shujian Zhou; Tingting Li; Ming Zhang; Chen Chen; Xiaomin Gao; Chen Zhang; Cai Hu; Qishenng Zuo; Guohong Chen; Bichun Li

doi:10.1016/j.gene.2021.145760

Epigenetic modification cooperates with Zeb1 transcription factor to regulate Bmp4 to promote chicken PGCs formation

Gene. 2021 Aug 20:794:145760. doi: 10.1016/j.gene.2021.145760. Epub 2021 Jun 9.

Authors

Shujian Zhou¹, Tingting Li¹, Ming Zhang¹, Chen Chen¹, Xiaomin Gao¹, Chen Zhang¹, Cai Hu¹, Qishenng Zuo¹, Guohong Chen¹, Bichun Li²

Affiliations

¹ Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, China.
² Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, China; College of Biotechnology, Jiangsu University of Science and Technology, China. Electronic address: yubcli@yzu.edu.cn.

PMID: 34116118
DOI: 10.1016/j.gene.2021.145760

Abstract

BMP4 is the critical gene of primordial germ cell formation in mammal, however, the mechanism of PGCs formation in chicken still unknown. In this research, we compared the evolution relationship of different species. Although the protein sequence is highly conservative between mouse, human and chicken, promotors vary among avian and mammal species. Therefore, it is easily to predict that there would be different regulation mechanism of Bmp4 expression in chicken. Here, we elucidate the function of chicken Bmp4 during PGCs formation. In vivo, Bmp4 can promote PGCs development and migration, and increase the expression of key genes (Cvh, c-kit, cxcr4, etc.). Whereas, the expression of these genes will decrease after knocking out Bmp4. After over-expression and knockout Bmp4 in vitro, we found that overexpression of Bmp4 could promote the formation of embryoid bodies (EB) and up-regulate the key genes of PGCs formation and migration, while knockout Bmp4 could inhibit the formation of embryoid bodies and decrease the expression of related genes. Flow and indirect immunofluorescence also indicated the same result. These all results proved that chicken Bmp4 could also promote the formation of PGCs. Furthermore, dual-luciferase activity detection showed that the promotor activity of Bmp4 was positively regulated by transcription factor Zeb1. Overexpression of Zeb1 can also increase the mRNA and protein expression of Bmp4. At the same time, DNA methylation inhibited Bmp4 transcription and histone methylation was able to promote its transcription. In conclusion, this study established that chicken Bmp4 can promote the formation of chicken PGCs. This gene is regulated by DNA, histone methylation and transcription factor Zeb1. These results lay a theoretical foundation for exploring the function and molecular mechanism of Bmp4 in the process of PGCs formation.

Keywords: Bmp4; Chicken; Expression regulation; PGCs; Promotor.

MeSH terms

Animals
Bone Morphogenetic Protein 4 / genetics*
Bone Morphogenetic Protein 4 / metabolism
Cell Movement
Cells, Cultured
Chickens
DNA Methylation
Embryoid Bodies / metabolism
Embryonic Germ Cells / cytology*
Embryonic Germ Cells / metabolism
Epigenesis, Genetic
Gene Expression Regulation, Developmental
Gene Knockout Techniques
Zinc Finger E-box-Binding Homeobox 1 / genetics*

Substances

Bone Morphogenetic Protein 4
Zinc Finger E-box-Binding Homeobox 1