Introduction:
The renewal potential of stem cells relies on their surrounding microenvironment. This involves the supporting growth factor milieu and cells. Sertoli cells (SCs) are the only adult cell in the seminiferous tubules that are essential for testis formation and spermatogenesis. Sertoli cells along with their secreted factors are vital components of the testis microenvironment, which advance the development of male germ stem cells to spermatozoa. Sertoli cells express the cytokine glial cell line-derived neurotrophic factor (GDNF) under the control of the follicle-stimulating hormone. GDNF is an essential regulator of SSC self-renewal and survival in vitro and is required for maintenance of the undifferentiated spermatogonial stem cells population in vivo. Vimentin plays an important role to maintain cell shape, cell motility, and intracellular trafficking along with other cytoskeletal protein during spermatogenesis. Methotrexate (MTX) is an anti-metabolite widely used in the treatment of neoplastic disorders, rheumatoid arthritis and psoriasis. The present study explored the mechanism of cytotoxic and genotoxic effects of MTX in a primary culture of Sertoli cells in vitro. DNA damage was evaluated using the Comet assay. The mRNA expression and their proteins of GDNF and vimentin as well as P53 and ATM genes and their proteins involved in apoptosis were also investigated using quantitative polymerase chain reaction (qPCR) and Western blot methods.
Conclusion
Results of the present study clearly showed that MTX-induced DNA damage as evident from the different Comet assay parameters. The GDNF and vimentin disruption seen in our in vitro study correlates with the increased activation of p53 and ATM in cells. However, the Sertoli cells appear to be physically supporting the spermatogenic cells, even though they may have lost their connection with vimentin. We can only hypothesize that MTX affects the Sertoli cells, causing immediate damage to the vimentin filaments and GDNF. Failure in any of these steps during endocytosis via Sertoli cells can lead to structural and junctional cell changes, resulting in an unstable environment for spermatogenesis.
Biography
Diana Anderson (H index 64) holds the Established Chair in Biomedical Sciences at the University of Bradford. She obtained her first degree in the University of Wales and second degrees in the Faculty of Medicine, University of Manchester. She has 500+ peer-reviewed papers, 10 books, has successfully supervised 32 PhDs and 4 research Master degrees, is an Editorial Board Member of 10 international journals. She is Editor-in-Chief of a book series on Toxicology for the Royal Society of Chemistry. She gives plenary and key note addresses at various international meetings. She is a consultant for many international organizations, including WHO, EU, NATO, TWAS, UNIDO and OECD.
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