Influence of Glutathione on Kinetic Parameters of Frozen-Thawed Spermatozoa from Ovchepolian Pramenka Rams

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Motility patterns of spermatozoa are an indicator of their fertilizing capacity. Reduced glutathione (GSH) has been reported to induce positive effects on the biological quality of the frozen-thawed spermatozoa. The aim of this study was to investigate the effects of GSH addition to semen extender based on the following kinetic parameters: continuous line velocity (VSL), average path velocity (VAP), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), linearity (LIN), straightness (STR), beat cross frequency (BCF), total motility (tMOT) and progressive motility (pMOT), and to appoint them as indicators of its presence. A soybean-based extender was used for dilution of the semen samples, fractioned in two parts, one containing GSH (5 mMol/ml) and second without GSH. The ejaculates (n=48) were collected from two rams (January - May, 2013) which were classified in two groups according to the used extender: Group 1 (with GSH, n=24) and Group 2 (without GSH, n=24), and then frozen in liquid nitrogen on -196°C degrees in a programmable freezer. Assessment of the samples has been performed post-thawing (30 sec. at 37°C) on CASA equipment, acquiring kinetic parameters. Results showed that only VSL and BCF have a statistically significant difference between group1 and group 2 (102.98±15.13 vs. 88.47±20.63, t=2.77, p<0.01 and 32.01±2.68 vs. 89.47±2.92, t=3.13, p<0.01, respectively). The summary of the investigation concludes that none of the kinetic parameters could be appointed as indicators that confer the positive effect of GSH as an additive to soy-bean semen extender.

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  • 1. Nikolovski M. Atanasov B. Dovenska M. Petkov V. Dovenski T. (2012). Variations in semen quality parameters of Ovchepolian Pramenka rams according to the method of collection and the meteorological season. Mac Vet Rev 35(2): 79-86.

  • 2. Salamon S. Maxwell W. M. C. (2000). Storage of Ram Semen. Animal Reproduction Science 62 77-111.

  • 3. Aitken R.J. (1984). Pathophysiology of human spermatozoa. Curr. Opin. Obstet. Gyn. 6 128-135.

  • 4. Maxwell W.M.C. Watson P.F. (1996). Recent progress in the preservation of ram semen. Anim. Reprod. Sci. 42 55-65.

  • 5. Bucak M.N. Atessahin A. Yuce A. (2008). Effect of anti-oxidants and oxidative stress parameters on ram semen after the freeze-thawing process. Small Ruminant Research 75 128-134.

  • 6. Alvarez J.G. Storey B.T. (1982). Spontaneous lipid peroxidation in rabbit epididymal spermatozoa: its effect on sperm motility. Biol. Reprod. 27 1102-1108. PMid:6297627

  • 7. Alvarez J.G. Storey B.T. (1984). Assessment of cell damage caused by spontaneous lipid peroxidation in rabbit spermatozoa. Biol. Reprod. 30 833-841. PMid:6231058

  • 8. Baumber J. Ball B.A. Gravance C.G. Medina V. Davies-Morel M.C.G. (2000). The effect of reactive oxygen species on equine sperm motility viability acrosomal integrity mitochondrial membrane potential and membrane lipid peroxidation. J. Androl. 21 895-902. PMid:11105916

  • 9. Nikolovski M. Petkov V. Trojacanec P. Milovanovic A. Barna A. Atanasov B. Dovenski T. (2012). Cryopreservation of Ovchepolean Pramenka ram semen with different diluents and antioxidants. SEE-ERA.NET PLUS framework project 083 (INDI_SHEEP TRADI_CHEESE); Final Scientifi c Conference Thermi Thessaloniki Greece 2012; Sustainable production of cheeses from local sheep milk in the Balkans; Book of abstracts pp.13

  • 10. de Lamirande E. Gagnon C. (1992). Reactive oxygen species and human spermatozoa. I. Effects on the motility of intact spermatozoa and on sperm axonemes. J. Androl. 13 368-378. PMid:1331006

  • 11. Griveau J. F. Le Lannou D. (1994). Effects of antioxidants on human sperm preparation techniques. Int. J. Androl. 17 225-231. PMid:7698847

  • 12. Lenzi A. Gandini L. Picardo M. (1998). A rationale for glutathione therapy. Hum Reprod.13(6):1419-22. PMid:9688363

  • 13. Irvine D.S. (1996). Gluthatione as a treatment for male infertility. Rev.Reprod. 1 6-12. PMid:9414432

  • 14. Watson PF. (1995). Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reproduction Fertility and Development 7 871-891. Mid:8711221

  • 15. Mortimer S. (1997). A critical review of the physiological importance and analysis of sperm movement in mammals. Human Reproduction Update 3 5 403-439. PMid:9528908

  • 16. E. Del Olmo A. Bisbal A. Maroto-Morales O. Garcia-Alvarez M. Ramon P. Jimenez-Rabadan F. Martinez-Pastor A.J. Soler J.J. Garde M.R. Fernandez-Santos (2013). Fertility of cryopreserved ovine semen is determined by sperm velocity. Animal Reproduction Science 138; 102- 109. PMid:23497922

  • 17. Mortimer D. (1990). Objective analysis of sperm motility and kinematics. In: Handbook of the Laboratory Diagnosis and Treatment of Infertility. (pp. 97-133). Keel B.A. and WebsterB.W. (eds) CRC Press Boca Raton

  • 18. Papadopoulos S. Hanrahan J.P. Donovan A. Duffy P. Boland M.P. Lonergan P. (2005). In vitro fertilization as a predictor of fertility from cervical insemination of sheep. Theriogenology 63150-159. PMid:15589281

  • 19. Rodriguez-Martinez H. (2003). Laboratory semen assessment and prediction of fertility: still utopia. Reprod. Domest. Anim. 38 312-318. PMid:12887570

  • 20. Schneider C.S. Ellington J.E. Wright Jr. R.W. (1999). Relationship between bull field fertility and in vitro embryo production using sperm preparation methods with and without somatic cell co-culture. Theriogenology 51 1085-1098.

  • 21. Zhang B.R. Larsson B. Lundeheim N. Rodriguez- Martinez H. (1998). Sperm characteristics and zona pellucida binding in relation to field fertility of frozen-thawed semen from dairy AI bulls. Int. J. Androl. 21 207-216. PMid:9749351

  • 22. Soler A.J. Poulin N. Fernandez-Santos M.R. Cognie Y. Esteso M.C.Garde J.J. Mermillod P. (2008). Heterologous in vitro fertility evaluation of cryopreserved Iberian red deer epididymal spermatozoa with zona-intact sheep oocytes and its relationship with the characteristics of thawed spermatozoa. Reprod. Domest. Anim. 43 293-298. PMid:18067534

  • 23. Gillan L. Evans G. Maxwell W.M.C. (2003). Flow cytometric evaluation of sperm parameters in relation to fertility potential. Theriogenology 63 445-457. PMid:15626410

  • 24. Gualtieri R. Boni R. Tosti E. Zagami M. Talevi R. (2005). Intracellular calcium and protein tyrosine phosphorylation during the release of bovine sperm adhering to the fallopian tube epithelium in vitro. Reproduction 129 51-60. PMid:15615898

  • 25. Garsia-Alvarez O. Maroto-Morales A. Martinez- Pastor F. Fernandez-Santos M.R. Esteso M.C. Pirez-Guzman M.D. Soler A.J. (2009). Heterologous in vitro fertilization is a good procedure to assess the fertility of thawed ram spermatozoa. Theriogenology 71 643-650. PMid:18996578

  • 26. O’Meara C.M. Hanrahan J.P. Prathalingam N.S. Owen J.S. Donovan A. Fair S. Ward F. Wade M. Evans A.C. Lonergan P. (2008). Relationship between in vitro sperm functional tests and in vivo fertility of rams following cervical artificial insemination of ewes with frozen-thawed semen. Theriogenology 69 513-522. PMid:18248736

  • 27. Garcia-Alvarez O. Maroto-Morales A. Ramуn M. del Olmo E. MontoroV. Dominguez- Rebolledo A.E. Bisbal A. Jiminez-Rabadan P. Pirez-Guzman M.D. Soler A.J. (2010). Analysis of selected sperm by density gradient centrifugation might aid in the estimation of in vivo fertility of thawed ram spermatozoa. Theriogenology 74 979-988. PMid:20580077

  • 28. Malo A.F. Garde J.J. Soler A.J. Garcia A.J. Gomendio M. Roldan E.R. (2005). Male fertility in natural populations of red deer is determined by sperm velocity and the proportion of normal spermatozoa. Biol.Reprod. 72 822-829. PMid:15576823

  • 29. Gage M.J. Macfarlane C.P. Yeates S. Ward R.G. Searle J.B. Parker G.A. (2004). Spermatozoal traits and sperm competition in Atlantic salmon: relative sperm velocity is the primary determinant of fertilization success. Curr. Biol. 14 44-47. PMid:14711413

  • 30. Holt W.V. Shenfield F. Leonard T. Hartman T.D. North R.D. Moore H.D. (1989). The value of sperm swimming speed measurements in assessing the fertility of human frozen semen. Hum. Reprod. 4 292-297. PMid:2715304

  • 31. Anel-Lopez L. Alvarez-Rodriguez M. Garcia- Alvarez O. Alvarez M. Maroto-Morales A. Anel L. de Paz P. Garde J.J. Martinez-Pastor F. (2012). Reduced glutathione and Trolox (vitamin E) as extender supplements in cryopreservation of red deer epididymal spermatozoa. Anim. Reprod. Sci. 135; (1-4): 37-46.

  • 32. Animal protection and welfare act 113/07. Official Gazette of the Republic of Macedonia. 20.09.2007.

  • 33. AshrafiI. Kohram H. Naijian H. Poorhamdollah M. Mirzakhani H. (2011). Effect of controlled and uncontrolled cooling rate on motility parameters of cryopreserved ram spermatozoa. African Journal of Biotechnology 10(44) 8965-8969.

  • 34. Mortimer S.T. Maxwell W.M.C. (2004). Effect of medium on the kinematics of frozen-thawed ram spermatozoa. Reproduction 127 285-291. PMid:15056794

  • 35. Dovenski T. Trojacanec P. Petkov V. Popovska- Percinic F. Dovenska M. Atanasov B. Ilievska K. Mickov L. Nikolovski M. (2012). Laparocscopic intrauterine artificial insemination by deep-frozen/ thawed ram semen from Ovchepolean Pramenka breed. SEE-ERA.NET PLUS framework project 083 (INDI_SHEEP TRADI_CHEESE); Final scientific conference Thermi Thessaloniki Greece; Sustainable production of cheeses from local sheep milk in the Balkans; Book of abstracts pp. 12

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