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Correlation Between Alkaline Phosphatase Expression and Sox2, Oct4, and Nanog Genes in Spermatogonial and ES-Like Cells

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Abstract

Purpose

The signaling pathway of spermatogenesis may be identified by analyzing and contrasting the similarities and differences in the expression and regulation of important pluripotent stem cell genes during spermatogenesis.

Methods

In this study, after isolating spermatogonial, embryonic stem cells and embryonic stem–like cells (ES-like) were prepared. The expression of alkaline phosphatase is then measured, and then it was discovered how genes directly connected to alkaline phosphatase interact by investigating their protein-protein interaction network. In the following, Fluidigm PCR and immunocytochemistry are utilized in spermatogonial, embryonic, and ES-like stem cells to evaluate the expression of Sox2, Oct4, and Nanog genes.

Results

Alkaline phosphatase is expressed positively in spermatogonial stem cells and pluripotent stem cells but not in Sertoli cells or fibroblasts, indicating that this enzyme is exclusive to these cell types. Sox2, Oct4, and Nanog have a positive and significant expression (P<0.05) in spermatogonial, embryonic stem cells, and ES-like cells, and a decreased expression is observed in differentiated and Sertoli cells.

Conclusion

Alkaline phosphatase and the genes Sox2, Oct4, and Nanog, which have a critical role in regulating the spermatogenesis process, have a regulatory role on each other’s activity and have a specific expression pattern in pluripotent and spermatogonial stem cells.

Lay Summary

The expression of Sox2, Oct4, and Nanog genes are very vital in regulating the expression of other genes and factors involved in the control and regulation of spermatogenesis. Therefore, examining their expression level and also how they are related can be helpful in the finding causes of infertility diseases. Also, investigating the sources of pluripotent stem cells like ES-like cells can be very important in relation to the emergence of new cell therapy methods.

Graphical Abstract

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Data Availability

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary materials.

Abbreviations

Oct4:

octamer-binding transcription factor 4

Sox2:

Sry-related box-2

IMH:

immunohistochemistry

ICC:

immunocytochemistry

SSCs:

spermatogonial stem cells

PTMs:

peritubular myoid cells

SCs:

Sertoli cells

PGCs:

primordial germ cells

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Acknowledgements

The faculty of biotechnology of Amol University of Special Modern Technologies, Islamic Republic of Iran, and the University of Heidelberg, Institute supported this research for Anatomy and Cell Biology III, Department of Neuroanatomy, Germany.

Funding

This research has been done with the financial support of the Centre for International Scientific Studies and Collaboration (CISSC) of the Ministry of Science, Research and Technology as well as the Iranian National Science Foundation (INSF) under project No.4006239, and a memorandum of understanding (MOU) agreement between the Institute for Anatomy and Cell Biology at the University of Heidelberg and Amol University of Special Modern Technology.

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Authors and Affiliations

  1. Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol, Iran

    Emad Reza & Hossein Azizi

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  1. Emad Reza
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  2. Hossein Azizi
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Contributions

ER: designed the bioinformatics data, wrote the manuscript, and did the data analysis. HA: accomplished and design the experiment, assembly of data, and data analysis. The authors read and approved the final manuscript for publication.

Corresponding author

Correspondence to Hossein Azizi.

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Ethical Consideration

In the current investigation, animal experiments were approved (Ir.ausmt.rec.1400.29.) by the Ethics Committee of Amol University of Special Modern Technologies.

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The authors declare no competing interests.

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Description of Future Works

In the future, it is targeted that by knowing the relationship and effects of factors and also by examining the differentiation conditions of spermatogonial stem cells or pluripotent sources such as ES-like cells, mature and fertile sperm cells can be created for the treatment of infertility.

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Reza, E., Azizi, H. Correlation Between Alkaline Phosphatase Expression and Sox2, Oct4, and Nanog Genes in Spermatogonial and ES-Like Cells. Regen. Eng. Transl. Med. (2023). https://doi.org/10.1007/s40883-023-00316-y

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  • DOI: https://doi.org/10.1007/s40883-023-00316-y

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