Pleiotropic Effect of Cyclophosphamide on Key Factors in Spermatogenesis Produced by Sertoli and Peritubular Cells of Immature Mice

Ronnie Solomon ¹ Mahmoud Huleihel ⁴ Ali AbuMadigem ² Eitan Lunenfeld ³

¹The Shraga Segal Dep. of Microbiology, Immunology and Genetics, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev
³Dep. OB/GYN, Soroka Medical Center, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev
⁴The Shraga Segal Dep. of Microbiology, Immunology and Genetics, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, The National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev

Introduction: Sertoli cells (SCs) produce key factors that affect spermatogenesis: Glial-derived nerve growth factor (GDNF), stem cell factor (SCF), leukemia inhibitory factor (LIF), colony stimulating factor-1 (CSF-1), inhibin B, androgen binding protein (ABP) and transferrin (Tf). Peritubular cells (PTCs) produce some of these factors. Cyclophosphamide (CP) affects proliferating cells, including spermatogonial cells.

Aims: To examine the effect of CP on SCs and PTCs that produce the above factors in seminiferous tubules (STs) of immature mice.

Materials and methods: Seven-day-old mice were injected intraperitoneal with 100 mg/kg CP once a week during 3 weeks, or with saline (control). Mice were sacrificed 10 days after the last injection. Cells isolated from the STs were double immunofluorescence stained with antibodies (Abs) against vimentin (SCs) or α-smooth muscle actin (PTCs) and Abs against GDNF, SCF, CSF-1, LIF. The expression of these factors and of inhibin B, ABP, Tf were examined by qPCR.

Results: Our results show a significant increase in the expression of inhibin B, FSH receptor, Tf in cells of the STs of CP-treated mice compared to control. However, there was a significant reduction in the number of SCs and PTCs that produced GDNF, LIF, SCF and CSF-1 but not in the percentage of these cells.

Conclusions: This is the first study that shows a direct effect of CP on the activity and number of somatic cells of STs in immature mice. These results may deepen our understanding the mechanisms of chemotherapy on the functionality of somatic cells in testis of immature mice.