DNA fragmentation and immunohistochemical investigation of sperm in varicocele cases

Abstract Aim: The primary objective of this study is to evaluate the levels of DNA fragmentation in sperm in cases of varicocele and to detect potential alterations in sperm cells using immunohistochemical methods. The research aims to microscopically examine the effects of varicocele on spermatogenesis and the integrity of sperm DNA. Materials and Methods: In this study, semen samples collected with consent from infertile males aged 25-40 were analyzed for the presence of varicocele and other identified factors and were divided into two groups: a normozoospermic control group and a group diagnosed with varicocele. Direct swim-up techniques were employed to separate highly motile spermatozoa, thereby facilitating a detailed analysis of DNA fragmentation and sperm morphology. Sperm viability was assessed using Eosin Y staining, DNA fragmentation was evaluated with Acridine Orange staining methods, and sperm morphology was examined using Spermac staining kits. Protein expression was visualized through immunohistochemical staining methods, and the results were recorded using high magnification photography. Results: This research was conducted on sperm samples collected from males diagnosed with varicocele and a healthy control group at the Urology Department of Dicle University Medical Faculty. The findings indicate that the presence of varicocele in sperm cells is associated with reduced or negative expression of dynein and aquaporin proteins. Immunohistochemical analyses have revealed a diminished or absent expression of dynein proteins and aquaporins 3 and 5 in varicocele patients compared to healthy individuals. This suggests a potential disruption in sperm motility and water transfer mechanisms, contributing to a deeper understanding of the effects of varicocele on male fertility. Conclusion: The outcomes of this study unequivocally reveal the deleterious effects of varicocele disease on the structural and functional integrity of spermatozoa. Given that the dynein and aquaporin protein families are crucial for the movement and fluid balance of spermatozoa, the reduced expression or absence of these proteins sheds light on the molecular mechanisms of male infertility. Keywords: Sperm DNA fragmentation, immunohistochemistry, varicocele, protein expression, male infertility