Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
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Effect of Ethylene Glycol and Antioxidant Combination on Function of Frozen-thawed Spermatozoa in Korean Jeju Black Bull

Ethylene glycol과 항산화제가 제주흑우 동결정액에 미치는 영향

  • Ko, Min-Hee (College of Veterinary Medicine, Jeju National University) ;
  • Seo, Jong-Pil (College of Veterinary Medicine, Jeju National University) ;
  • Kang, Tae-Young (College of Veterinary Medicine, Jeju National University)
  • 고민희 (제주대학교 수의과대학) ;
  • 서종필 (제주대학교 수의과대학) ;
  • 강태영 (제주대학교 수의과대학)
  • Received : 2017.01.09
  • Accepted : 2017.02.21
  • Published : 2017.04.28
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Abstract

We investigated the effect of ethylene glycol and antioxidants such as taurine, hypotaurine and trehalose with extenders during cryopreservation of Korean Jeju Black Bull spermatozoa. The cryopreservation of freshly collected spermatozoa was conducted with four different conditions. As a control, spermatozoa were cryopreserved with Tris egg-yolk extenders added 5% ethylene glycol (EG). Taurine (20 mM), hypotaurine (20 mM) and trehalose (20 mM) were individually added into tris egg-yolk extenders with 5% EG. After thawing of frozen spermatozoa with four different conditions, sperm viability, motility, acrosomal integrity, and membrane integrity were investigated. The significant (p < 0.05) improvement of sperm viability showed in all antioxidant treated thawed spermatozoa (taurine; $68.1%{\pm}4.4$, hypotaurine; $69.2%{\pm}6.7$ and trehalose; $68.0%{\pm}4.4$) when compared to control ($63.4%{\pm}5.6$). Neither positive nor detrimental effects of three antioxidants were shown sperm motility after thawing. The results of hypo-osmotic swelling test showed that the membrane integrity of taurine, hypotaurine or trehalose treated thawed spermatozoa ($64.1%{\pm}5.4$, $61.5%{\pm}3.7$ and $59.0%{\pm}4.0$, respectively) had significantly (p < 0.05) higher rate of the swollen sperm compared to control ($53.7%{\pm}9.7$). Hypotaurine treated frozen-thawed spermatozoa had siginificantly higher (p < 0.05) F pattern ratio than taurine, trehalose and control treated frozen-thawed spermatozoa. Trehalose added frozen-thawed spermatozoa had significantly higher (p < 0.05) acrosome reaction pattern ratio than taurine and hypotaurine added frozen-thawd spermatozoa. In this study, we found that antioxidants such taurine, hypotaurine and trehalose treatments during cryopreservation process could reduce damage of spermatozoa of Korean Jeju Black Bull and improved sperm capability of fertilization.

Keywords

References

  1. 최두석, 문신용, 장윤석. 남성 불임검사 중 정자 형태와 정자 운동성 검사 및 저장성용액 내 정자 팽창검사의 상관관계 및 가임능력 예측에 관한 연구. 대한산부인과학회지 1993; 36: 2497-2509.
  2. 최선호, 고민희, 강태영, 조상래, 박용상, 오신애. 제주흑우 동결정액 제조시 ethylene Glycol의 농도와 예비 동결조건이 정자의 생존율 및 첨체양상에 미치는 영향. 한국동물번식학회지. 2011; 35: 377-383.
  3. Adeoya-Osiguwa SA, raser LR. Environmental estrogens and sperm function. Hum Reprod 2004; 19: 216-217. https://doi.org/10.1093/humrep/deh015
  4. Aisen EG, Alvarez HL, Venturino A, Garde JJ. Effect of trehalose and EDTA on cryoprotective action of ram semen diluents. Theriogenology 2000; 53: 1053-1061. https://doi.org/10.1016/S0093-691X(00)00251-X
  5. Aitken RJ, Gordon E, Harkiss D, Twigg JP, 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: 1037-1046. https://doi.org/10.1095/biolreprod59.5.1037
  6. Bilodeau JF, Chatterjee S, Sirard MA, agnon C. Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Mol Reprod Dev 2000; 55: 282-288. https://doi.org/10.1002/(SICI)1098-2795(200003)55:3<282::AID-MRD6>3.0.CO;2-7
  7. Bucak MN, Tuncer PB, Sariozkan S, Uluta PA, Coyan K, Ba pinar N, Ozkalp B. Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen. Res Vet Sci 2009; 87: 468-472. https://doi.org/10.1016/j.rvsc.2009.04.014
  8. Buhr MM, Fiser P, Bailey JL, Curtis EF. Cryopreservation in different concentration of glycerol alters boar sperm and their membranes. J Androl 2001; 22: 961-969. https://doi.org/10.1002/j.1939-4640.2001.tb03436.x
  9. Chenier T, Merkies K, Leibo S, Plante C, Johnson W. Evaluation of cryoprotective agents for use in the cryopreservation of equine spermatozoa. Proc. 44th Annual Convention of the American Association of Equine Practitioners, Baltilmore, Maryland, USA, 1998: 5-6.
  10. Fraser LR, Abeydeera LR, Niwa K. Ca2+-regulating mechanisms that modulate bull sperm capacitation and acrosome alexocytosis as determined by chlortetracycline analysis. Mol Reprod Dev 1995; 40: 233-241. https://doi.org/10.1002/mrd.1080400213
  11. Gilmore JA, Liu J, Woods EJ, Peter AT, Critser JK. Cryoprotective agent and temperature effects on human sperm membrane permeabilities: convergence of theoretical and empirical approaches for optimal cryopreservation methods. Hum Reprod 2000; 15: 335-343. https://doi.org/10.1093/humrep/15.2.335
  12. Gilmore JA, McGann LE, Liu J. Gao DY, Peter AT, Kleinhans FW, Critser JK. Effect of cryoprotectant solutes on water permeability of human spermatozoa. Biol Reprod 1995; 53: 985-995. https://doi.org/10.1095/biolreprod53.5.985
  13. Guthrie HD, Liu J, Critser JK. Osmotic tolerance limits and effects of cryoprotectants on motility of bovine spermatozoa. Biol Reprod 2002; 67: 1811-1816. https://doi.org/10.1095/biolreprod67.6.1811
  14. Hammerstedt RH, Graham JK, Nolan JP. Cryopreservation of mammalian sperm: what we ask them to survive. J Androl 1990; 11: 73-88.
  15. Hu JH, Li QW, Jiang ZL, Yang H, Zhang SS, Zhao HW. The cryoprotective effect of trehalose supplementation on boar spermatozoa quality. Reprod Dom Anim 2009; 44: 571-575. https://doi.org/10.1111/j.1439-0531.2007.00975.x
  16. Jeyendran RS, Van der Ven HH, Prez-Pelaez M, Carbo BG, Zaneveld LJ. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J Reprod Fertil 1984; 70: 219-228. https://doi.org/10.1530/jrf.0.0700219
  17. Kommisrud E, Paulenz H, Sehested E, Greule IS. Influence of boar and semen parameters on motility and acrosome integrity in liquid boar semen stored for 5 days. Acta Vet Scand 2002; 43: 49-55. https://doi.org/10.1186/1751-0147-43-49
  18. Li G, Saenz J, Godke RA, Devireddy RV. Effect of GLY and cholesterol-loaded cyclodextrin on freezing-induced water loss in bovine spermatozoa. Reproduction 2006; 131: 875-886. https://doi.org/10.1530/rep.1.00995
  19. Mantovani R, Rota A, Falomo ME, Bailoni L, Vincenti L. Comparison between glycerol and ethylene glycol for the cryopreservation of equine spermatozoa: sperm quality assessment with standard analyses and with the hypoosmotic swelling test. Reprod Nutr Dev 2002; 42: 217-226. https://doi.org/10.1051/rnd:2002020
  20. Martins-Bessa A, Rocha A, Mayenco-Aguirre A. Effects of taurine and hypotaurine supplementation and ionophore concentrations on post-thaw acrosome reaction of dog spermatozoa. Theriogenology 2009; 71: 248-253. https://doi.org/10.1016/j.theriogenology.2008.07.006
  21. McGonagle LS, Goldstein M, Feldschuh J, Foote RH. The influence of cryoprotective media and processing procedures on motility and migration of frozen-thawed human sperm. Asian J Androl 2002; 4: 137-141.
  22. Oh SA, Park YJ, You YA, Mohamed EA, Pang MG. Capacitation status of stored boar spematozoa is related litter size of sows. Anim Reprod Sci 2010; 121: 131-138. https://doi.org/10.1016/j.anireprosci.2010.05.019
  23. Parker NA, Bailey TL, Bowen JM, Ley WB, Purswell BJ, Dascanio JJ. In vitro and xenogenous capacitation-like changes of fresh, cooled, and cryopreserved stallion sperm as assessed by a chlortetracycline stain. J Androl 2000; 21: 45-52.
  24. Pena AI, Lugilde LL, Barrio M, Herradon PG, Quintela LA. Effects of Equex from different sources on post-thaw survival, longevity and intracellular Ca2+ concentration of dog spermatozoa. Theriogenology 2003; 59: 1725-1739. https://doi.org/10.1016/S0093-691X(02)01233-5
  25. Pereira SM, Rigon RM, Mezzalira A, Cecim M. Ethylene glycol on canine semen cryopreservation. Ciencia Rural 2002; 32: 649-655. https://doi.org/10.1590/S0103-84782002000400017
  26. Sariozkan S, Bucak MN, Tuncer PB, Ulutas PA, Bilgen A. The influence of cysteine and taurine on microscopicoxidative stress parameters and fertilizing ability of bull semen following cryopreservation. Cryobiology 2009; 58: 134-138. https://doi.org/10.1016/j.cryobiol.2008.11.006
  27. Silva AR, de Cassia Soares Cardoso R, Uchoa DC, MacHado da Silva LD. Effect of Tris-buffer, egg yolk and GLY on canine semen freezing. Vet J 2002; 164: 244-246. https://doi.org/10.1053/tvjl.2002.0704
  28. Shiva Shankar Reddy N, Jagan Mohanarao G, Atreja SK. Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation. Anim Reprod Sci 2010; 119: 183-190. https://doi.org/10.1016/j.anireprosci.2010.01.012
  29. Squires EL, Keith SL, Graham JK. Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology 2004; 62: 1056-1065. https://doi.org/10.1016/j.theriogenology.2003.12.024
  30. Storey BT Biochemistry of the induction and prevention of lipoperoxidative damage in human spermatozoa. Mol Hum Reprod 1997; 3: 203-213. https://doi.org/10.1093/molehr/3.3.203
  31. Suzuki K, Geshi M, Yamauchi N, Nagai T. Functional changes and motility characteristics of Japanese Black Bull spermatozoa separated by Percoll. Anim Reprod Sci 2003; 77: 157-172. https://doi.org/10.1016/S0378-4320(03)00035-6
  32. Yanagimachi R, Yanagimachi H, Rogers BJ. The use of zonafree animal ova as a test-system for the assessment of the fertilizing capacity of human spermatozoa. Biol Reprod 1976; 15: 471-476. https://doi.org/10.1095/biolreprod15.4.471