Integration of RRBS and RNA-seq unravels the regulatory role of DNMT3A in porcine Sertoli cell proliferation

Dong Xu; Saina Yan; Huimin Jin; Chujie Chen; Xiangwei Tang; Xu Wang; Yonghong Li; Fang Fei; Anqi Yang

doi:10.3389/fgene.2023.1302351

Integration of RRBS and RNA-seq unravels the regulatory role of DNMT3A in porcine Sertoli cell proliferation

Front Genet. 2024 Jan 9:14:1302351. doi: 10.3389/fgene.2023.1302351. eCollection 2023.

Authors

Dong Xu^#¹, Saina Yan^#², Huimin Jin², Chujie Chen³, Xiangwei Tang³, Xu Wang³, Yonghong Li¹, Fang Fei¹, Anqi Yang²

Affiliations

¹ Department of Biological and Environmental Engineering, Yueyang Vocational Technical College, Yueyang, Hunan, China.
² School of Basic Medical Sciences, Hengyang Medical School, University of South China, Hengyang, Hunan, China.
³ College of Animal Science and Technology, Hunan Agricultural University, Changsha, Hunan, China.

^# Contributed equally.

Abstract

DNMT3A participates in de novo methylation, yet its impact on the proliferation of testicular Sertoli cells remains unclear. Development-specific methylation has been proven to be associated with cellular development. Therefore, in this study, we simulated DNMT3A expression pattern during testicular development by DNMT3A interference. Then, RRBS and RNA-seq were used to decipher DNMT3A regulatory mechanisms on Sertoli cell proliferation. Immunofluorescence staining revealed the expression of DNMT3A in the Sertoli cells of the prepubertal testis. DNMT3A was demonstrated to inhibit the cell cycle and proliferation of Sertoli cells, while promoting cell apoptosis. After transfected with DNMT3A interference, a total of 560 DEGs and 2,091 DMGs produced by DNMT3A interference were identified between two treated groups, respectively. Integrating the results from RRBS and RNA-seq, the overlapping genes between DMGs and DEGs were found to be enriched in the Gene Ontology (GO) terms related to cellular development and the Apelin signaling pathway. The present study demonstrated the impact of DNMT3A on the proliferation of porcine testicular Sertoli cells, suggesting that DNMT3A primarily acts through the Apelin signaling pathway. These findings provide valuable insights into how DNMT3A influences testicular development and health, offering new perspectives.

Keywords: DNMT3A; RNA-seq; RRBS; Sertoli cells; proliferation.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was financially supported by the Science and Education Joint Funds of the Hunan Provincial Natural Science Foundation of China (2021JJ60009), the Hunan Provincial Department of Education Scientific Research Project (23B1097), and the Hengyang Municipal Agricultural Guidance Program (2023). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.