Quantitative proteomic analysis based on tandem mass tagging (TMT) reveals key proteins related to DNA hydroxymethylation enzyme TET1 for spermatogonia self-renewal

Abstract

Abstract Background Abnormal spermatogonia (SSCs) can cause spermatogenic disorders such as weak spermatozoa, oligospermia, and azoospermia. DNA hydroxymethylase TET1 hydroxylates the methylation sites of specific genes, enabling the process of DNA demethylation and regulating gene expression. However, the key differential genes affected by the specific action of TET1 and the mechanism of interaction between the differential genes are not clear. Result In this study, we applied quantitative proteomics techniques based on Tandem Mass Tags (TMT) to screen the 24h differentially expressed proteins in the TET1 overexpression group (MYC-TET1) and the control group (MYC) to provide a basis for studies such as the regulation of TET1-mediated epigenetic modifications on SSCs. By TMT technique, we identified 5891 proteins, of which 337 were significantly differentially expressed, 76 were up-regulated and 261 were down-regulated. Gene Ontology (GO) enrichment analysis revealed that proteins with significant differential expression such as RARG, RN114, DJC30, and ABHD2 were associated with functions such as sperm-egg recognition, sperm-egg fusion, sperm ejaculation, spermatogenesis and development, and embryonic development. changes in proteins such as GHR, CCNT1, HTRA1, and ANXA3 affected cell viability, gene transcription and translation activities, and important intracellular biological processes in SSCs. intracellular biological processes. Conclusions In this study, we obtained differential protein profiles by overexpressing TET1 in SSCs and subsequently by TMT protein sequencing technology, combined with Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis, GO enrichment analysis and protein interaction network map to jointly analyze the epistatic regulatory role of TET1 on SSCs, which provides a scientific basis for further study of spermatogenesis and contributes to the understanding of male reproductive system diseases.