Abstract
Purpose
In the female genital tract spermatozoa interact with the oviductal secretion. The aim of the study was to evaluate the effect of conditioned media (CM) from cultures of human oviductal tissue, on sperm DNA integrity. The effect of H2O2 on sperm DNA integrity, before and after incubation under capacitating conditions, was also investigated.
Methods
Motile sperm obtained from normozoospermic semen samples were incubated (4 h or 22 h) in the presence or absence of CM and further exposed to H2O2. DNA damage was detected by the comet assay.
Results
The CM significantly reduced the DNA damage associated with sperm incubation, and also decreased the effect of H2O2 after 4 h incubation, compared to controls. The H2O2 caused a dose-dependent deleterious effect on sperm DNA integrity both before and following 22 h of capacitation.
Conclusion
The oviductal tissue CM increased the stabilization of the sperm DNA structure under culture conditions.
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References
Agarwal A, Said T. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Updat 2003;9:331–45.
Saleh R, Agarwal A. Oxidative stress and male infertility: from research bench to clinical practice. J Androl 2002;23:737–52.
Kovalski NN, de Lamirande E, Gagnon C. Reactive oxygen species generated by human neutrophils inhibit sperm motility: protective effect of seminal plasma and scavengers. Fertil Steril 1992;58:809–16.
Kessopoulou E, Tomlinson MJ, Banat CLR, Bolton AE, Cooke ID. Origin of reactive oxygen species in human semen-spermatozoa or leukocytes. J Reprod Fertil 1992;94:463–70.
Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ Jr, Agarwal A. Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum Reprod 2001;16:1922–30.
Aitken RJ, Clarkson JS. Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques. J Androl 1988;9:367–76.
Aitken RJ, Fisher HM, Fulton N, Gomez E, Knox W, Lewis B, Irvine S. Reactive oxygen species generation by human spermatozoa is induced by exogenous NADPH and inhibited by the flavoprotein inhibitors diphenylene iodonium and quinacrine. Mol Reprod Dev 1997;47:468–82.
Gavella M, Lipovac V. NADH-dependent oxido-reductase (diaphorase) activity and isozyme pattern of sperm in infertile men. Arch Androl 1992;28:135–41.
Storey BT. Biochemistry of the induction and prevention of lipoperoxidative damage in human spermatozoa. Mol Hum Reprod 1997;3:203–14.
Agarwal A, Makker K, Sharma R. Clinical relevance of oxidative stress in male factor infertility: an update. Am J Reprod Immunol 2008;59:2–11.
Rich T, Allen RL, Wyllie AH. Defying death after DNA damage. Nature 2000;407:777–83.
Barzilai A, Yamamoto K. DNA damage responses to oxidative stress. DNA Repair 2004;3:1109–15.
Suarez S, Pacey A. Sperm transport in the female reproductive tract. Hum Reprod Updat 2006;12:23–37.
Troedsson MH, Desvousges A, Alghamdi AS, Dahms B, Dow CA, Hayna J, Valesco R, Collahan PT, Macpherson ML, Pozor M, Buhi WC. Components in seminal plasma regulating sperm transport and elimination. Anim Reprod Sci 2005;89:171–86.
Robertson SA. Seminal fluid signaling in the female reproductive tract: lessons from rodents and pigs. J Anim Sci 2007;85:E36–44.
Lea R, Sandra O. Immunoendocrine aspects of endometrial function and implantation. Reproduction 2007;134:389–404.
Hensley K, Robinson KA, Gabbita SP, Salsman S, Floyd RA. Reactive oxygen species, cell signaling, and cell injury. Free Radic Biol Med 2000;28:1456–62.
Zhu J, Barrat CL, Lippes J, Pacey AA, Cooke ID. The sequential effects of human cervical mucus, oviductal fluid, and follicular fluid on sperm function. Fertil Steril 1994;61:1129–35.
Kervancioglu ME, Djahanbakhch O, Aitken RJ. Epithelial cell coculture and the induction of sperm capacitation. Fertil Steril 1994;61:1103–8.
Yao Y, Ho P, Yeung WS. Human oviductal cells produce a factor(s) that maintains the motility of human spermatozoa in vitro. Fertil Steril 2000;73:479–86.
Quintero I, Ghersevich S, Caille AM, Munuce MJ, Daniele SM, Morisoli L. Effects of human oviductal in vitro secretion on spermatozoa and search of sperm-oviductal proteins interactions. Int J Androl 2005;28:137–43.
Munuce MJ, Serravalle A, Caille A, Zumoffen C; Botti G, Cabada M, Ghersevich S. Human tubal secretion can modify the affinity of human spermatozoa for the zona pellucida. Fertil Steril. 2008 (in press).
Lopes S, Jurisicova A, Sun J-G, Casper R. Reactive oxygen species: potential cause for DNA fragmentation in human spermatozoa. Hum Reprod 1998;13:896–900.
Duru NK, Morshedi M, Oehninger S. Effects of hydrogen peroxide on DNA and plasma membrane integrity of human spermatozoa. Fertil Steril 2000;74:1200–7.
Rivlin J, Mendel J, Rubinstein S, Etkovitz N, Breitbart H. Role of hydrogen peroxide in sperm capacitation and acrosome reaction. Biol Reprod 2004;70:518–22.
O’Flaherty C, de Lamirande E, Gagnon C. Reactive oxygen species modulate independent protein phosphorylation pathways during human sperm capacitation. Free Radic Biol Med 2006;40:1045–55.
Chang MC. Fertilizing capacity of spermatozoa deposited into the fallopian tubes. Nature 1952;168:697–98.
Yanagimachi R. Mammalian fertilization. In: Knobil E, y Neill J, editors. The physiology of reproduction. New York: Raven; 1994. p. 189–317.
Davis LG, Kuehl M, Battey JF. Basic methods in molecular biology. 2nd ed. Connecticut, USA: Appleton and Lange; 1994. p. 16–21.
World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. New York: Cambridge University Press; 1999.
Kruger TF, Menkveld R, Stander FS, Lombard CJ, Van den Merwe JP, Van Zyl JA. Sperm morphologic features as a prognosis factor of in vitro fertilization. Fertil Steril 1986;46:1118–23.
Singh NP, Danner DB, Tice RR, McCoy MT, Collins GD, Schneider EL. Abundant alkali labile sites in DNA of human and mouse sperm. Exp Cell Res 1989;184:461–70.
Hughes CM, Lewis SEM, McKelvey-Martin VJ, Thompson W. A comparison of baseline and induced DNA damage in human spermatozoa from fertile and infertile men using a modified comet assay. Mol Hum Reprod 1996;2:613–20.
Piperakis SM, Visvardis EE, Tassiou AM. Comet assay for nuclear DNA damage. Methods Enzymol 1999;300:184–94.
Singh NP, Muller CH, Berger RE. Effects of age on DNA double-strand breaks and apoptosis in human sperm. Fertil Steril 2003;80:1420–30.
Aitken J, Gordon E, Harkiss D, Twigg J, Milne P, Jennings Z, Irvine DS. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod 1998;59:1037–46.
Wang X, Sharma RK, Sikka SC, Thomas AJ Jr, Falcone T, Agarwal A. Oxidative stress is associated with increased apoptosis leading to spermatozoa DNA damage in patients with male factor infertility. Fertil Steril 2003;80:531–5.
Morris ID, Ilott S, Dixon L, Brison DR. The spectrum of DNA damage in human sperm assessed by single cell gel electrophoresis (comet assay) and its relationship to fertilization and embryo development. Hum Reprod 2002;17:990–8.
Zini A, Finelli A, Phang D, Jarvi K. Influence of semen processing technique on human sperm DNA integrity. Urology 2000;56:1081–4.
Hughes CM, Lewis SE, McKelvey-Martin VJ, Thompson W. The effects of antioxidant supplementation during Percoll preparation on human sperm DNA integrity. Hum Reprod 1998;13:1240–7.
Aitken RJ, Vernet P. Maturation of redox regulatory mechanisms in the epididymis. J Reprod Fertil 1998;53:109–18.
Acknowledgements
The present study was partially supported by grants from CONICET (PID 02153) and FONCYT (PICT 0115092).
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Robert, C., Caille, A., Zumoffen, C. et al. Effect of human oviductal in vitro secretion on human sperm DNA integrity. J Assist Reprod Genet 25, 263–270 (2008). https://doi.org/10.1007/s10815-008-9228-z
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DOI: https://doi.org/10.1007/s10815-008-9228-z