Investigating the expression and role of N-Myc in spermatogonial stem cells and male infertility

Abstract

Background

In order to sustain reproduction, this study aims to investigate the role of N-Myc in the regulation of spermatogonial stem cell differentiation and the consequent generation of sperm during spermatogenesis. By analyzing specific gene expression patterns associated with N-Myc signaling pathways, we seek to identify potential targets for infertility treatments.

Main methods

In this study on mice, we constructed a PPI network in embryonic stem cells, germ cells, SSCs and pluripotent stem cells using the GEO datasets, NCBI, STRING, and Cytoscape databases. Then, after isolating spermatogonial stem cells from 6 C57BL/6 mice, ES-like cells were prepared. We further examined N-Myc, Oct4, Sox9, Vasa, and Plzf expression in testes, SSCs, and ES-like cells by immunohistochemistry (IHC) and immunocytochemistry (ICC), also the expression of N-Myc using RT-PCR. Finally, we performed gene set enrichment and miRNA analysis for N-Myc using Enrichr and miRTarBase.

Findings

N-Myc has a central role in PPIs for genes maintaining pluripotency. In addition, this gene is coexpressed with Oct4, and positive expression of the Vasa and Plzf genes was observed in seminiferous tubule cells. The interaction network of N-Myc with microRNAs and lncRNAs was constructed, and the biological roles of N-Myc in spermatogenesis, such as glycolysis, were revealed.

Conclusion

The result highlight that N-Myc is essential for orchestrating the differentiation of spermatogonial stem cells into mature sperm by interacting with key regulatory factors. This underscores its pivotal role in both maintaining pluripotency and facilitating successful spermatogenesis, which may inform future strategies for addressing infertility.