Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Effect of Adding Taurine, Hypotaurine and Trehalose as Antioxidants to a Tris-based Egg Yolk Extender on Korean Jeju Black Bull Sperm Quality Following Cryopreservation

제주흑우 동결정액 제조에 있어 난황 Tris 희석제에 항산화제로서 Taurine, Hypotaurine 그리고 Trehalose의 첨가가 동결 융해 후 정자의 성상에 미치는 영향

  • 오신애 (농촌진흥청 국립축산과학원) ;
  • 고민희 (농촌진흥청 국립축산과학원) ;
  • 강태영 (농촌진흥청 국립축산과학원) ;
  • 최선호 (농촌진흥청 국립축산과학원) ;
  • 고문석 (농촌진흥청 국립축산과학원) ;
  • 정영호 (중부대학교 애완동물자원과학과) ;
  • 조원모 (농촌진흥청 국립축산과학원)
  • Received : 2012.07.10
  • Accepted : 2012.08.24
  • Published : 2012.08.31
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Abstract

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. The objective of this study was to investigate the effect of taurine, hypotaurine and trehalose as antioxidants on the function of the freezing-thawed sperm in Korean Jeju Black Bull. The semen was cryopreserved with tris egg yolk extendercontaining 7% glycerol and treated with 20mM taurine, hypotaurine and trehalose. Frozen-thawed sperms were evaluated for sperm motility, viability, membrane integrity, acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender containing 7% glycerol only as control. Frozen-thawed semen evaluation clearlyindicated that the addition of taurine or hypotaurine significantly improved (p<0.05) the motility and viability compared to control spermatozoa. Moreover, in membrane integrity, swollen sperm ratio was significantly increased (p<0.05) in taurine, hypotaurine or trehalose compared to control. In sperm acrosome integrity, F pattern ratio was increased (p<0.05) in hypotaurine among treatments, and AR pattern was significantly lowered (p<0.05) in taurine, hypotaurine and trehalose. In assessed sperm fertilizing ability, taurine, hypotaurine or trehalose significantly improved (p<0.05) the ratio of pronucleus formation and SFI. Finally, compared with the control, addition of taurine, hypotaurine or trehalose as an antioxidant to the freezing extender showed more positive effects on the frozen-thawed spermatozoa. It is concluded that the addition of taurine, hypotaurine, or trehalose to the freezing extender could reduce cryodamage of the Korean Jeju Black Bull spermatozoa.

제주흑우의 동결 정액 제조시 동결 보존액에 첨가한 taurine과 hypotaurine은 동결 융해 후 정자의 운동성, 생존성 그리고 정자막 온전성을 개선시키는 것을 알 수 있었다. Taurine과 hypotaurine의 첨가는 유의적으로 높은 운동성, 생존성을 보였으며(p<0.05), 특히 hypotaurine은 다른 실험구에 비하여 유의적으로 높은 정자막 온전성을 나타냈다(p<0.05). 뿐만 아니라, hypotaurine은 유의적으로 높은 F pattern 비율을 유지하였으며(p<0.05), 이들 항산화물질을 첨가한 실험구에서는 대조구에 비하여 유의적으로 낮은 AR pattern을 나타내어(p<0.05) 동결로 기인된 수정능획득 유사 상태 정자의 비율을 유의적으로 감소시켜 조기 첨체반응 비율을 감소시켰다. 정자의 난자내 침투능력에 있어서 모든 처리구에서 대조구보다 높은 웅성전핵 형성율과 SFI를 나타냈으며, hypotaurine의 처리는 가장 높은 침투능력을 나타냈으나 처리구간의 유의적 차이는 나타나지 않았다. 본 연구결과는 멸실위험의 토종가축 생식세포 및 유전자원 보존과 토종가축 육성을 위한 번식 증대를 위한 제주흑우 동결정액 제조에 중요한 이용 방법이 될 것이며, 동결 융해 후 정자의 기능 개선을 위한 다양한 희석제 및 첨가제를 활용한 연구가 필요할 것으로 사료된다.

Keywords

References

  1. Aboagla, E. M. and Terada, T. 2003. Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol. Reprod. 69:1245-1250. https://doi.org/10.1095/biolreprod.103.017889
  2. Aisen, E. G., Alvarez, H. L., Venturino, A. and Garde, J. J. 2000. Effect of trehalose and EDTA on cryoprotective action of ram semen diluents. Theriogenology 53:1053-1061. https://doi.org/10.1016/S0093-691X(00)00251-X
  3. Aisen, E., Quintana, M., Medina, V., Morello, H. and Venturino, A. 2005. Ultramicroscopic and biochemical changes in ram spermatozoa cryopreserved with trehalose-based hypertonic extenders. Cryobiology 50:239-249. https://doi.org/10.1016/j.cryobiol.2005.02.002
  4. Aitken, R. J., Clarkson, J. S. and Fishel, S. 1989. Generation of reactive oxygen species, lipid peroxidation, and human sperm function. Biol. Reprod. 41:183-197. https://doi.org/10.1095/biolreprod41.1.183
  5. Aitken, R. J., Buckingham, D. and Harkiss, D. 1993. Use of a xanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa. J. Reprod. Fertil. 97:441-450. https://doi.org/10.1530/jrf.0.0970441
  6. Aitken, R. J., Gordon, E., Harkiss, D., Twigg, J. P., Milne, P., Jennings, Z. and Irvine, D. S. 1998. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol. Reprod. 59:1037-1046. https://doi.org/10.1095/biolreprod59.5.1037
  7. Alvarez, J. G. and Storey, B. T. 1983. Taurine, hypotaurine, epinephrine and albumin inhibit lipid peroxidation in rabbit spermatozoa and protect against loss of motility. Biol. Reprod. 29:548-555. https://doi.org/10.1095/biolreprod29.3.548
  8. Alvarez, J. G. and Storey, B. T. 1989. Role of glutathione peroxidase in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation. Gamete. Res. 23:77-90. https://doi.org/10.1002/mrd.1120230108
  9. Anchordoguy, T. J., Rudolph, A. S., Carpenter, J. F. and Crowe, J. H. 1987. Modes of interaction of cryoprotectants with membrane phospholipids during freezing. Cryobiology 24:324-331. https://doi.org/10.1016/0011-2240(87)90036-8
  10. Bailey, J. L., Bilodeau, J. F. and Cormier, N. 2000. Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. J. Androl. 21:1-7.
  11. Bilodeau, J. F., Chatterjee, S., Sirard, M. A. and Gagnon, C. 2000. Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Mol. Reprod. Dev. 55:282-288. https://doi.org/10.1002/(SICI)1098-2795(200003)55:3<282::AID-MRD6>3.0.CO;2-7
  12. Boatman, D. E., Bavister, B. D. and Cruz, E. 1990. Addition of hypotaurine can reactivate immotile golden hamster spermatozoa. J. Androl. 11:66-72.
  13. Bolten, M., Weissbach, L. and Kaden, R. 2005. Cryopreserved human sperm deposits: Usability after decades of storage. Urologe A 44:904-908 https://doi.org/10.1007/s00120-005-0840-6
  14. Bucak, M. N., Atessahin, A., Varisli, O., Yuce, A., Tekin, N. and Akcay, A. 2007. The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen Microscopic and oxidative stress parameters after freeze-thawing process. Theriogenology 67: 1060-1067. https://doi.org/10.1016/j.theriogenology.2006.12.004
  15. Bucak, M. N., Tuncer, P. B., Sariozkan, S., Ulutaş, P. A., Coyan, K., Başpinar, N. and Ozkalp, B. 2009. Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen. Res. Vet. Sci. 87:468-472. https://doi.org/10.1016/j.rvsc.2009.04.014
  16. Chatterjee, S., de Lamirande, E. and Gagnon, C. 2001. Cryopreservation alters membrane sulfhydryl status of bull spermatozoa: protection by oxidized glutathione. Mol. Reprod. Dev. 60:498-506. https://doi.org/10.1002/mrd.1115
  17. Chen, Y., Foote, R. H., Tobback, C., Zhang, L. and Hough, S. 1993. Survival of bull spermatozoa seeded and frozen at different rates in egg yolk-tris and whole milk extenders. Dairy. Sci. 76:1028-1034. https://doi.org/10.3168/jds.S0022-0302(93)77430-5
  18. Donnelly, E. T., McClure, N. and Lewis, S. E. 2000. Glutathione and hypotaurine in vitro: effects on human sperm motility, DNA integrity and production of reactive oxygen species. Mutagenesis 15:61-68. https://doi.org/10.1093/mutage/15.1.61
  19. Eaglesome, M. D. and Garcia, M. M. 1990. The effect of Mycoplasma bovis on fertilization processes in vitro with bull spermatozoa and zona-free hamster oocytes. Vet. Microbiol. 21:329-337. https://doi.org/10.1016/0378-1135(90)90004-F
  20. Foote, RH., Chen Y., Brocket't C. C. and Kaproth, M. T. 1993. Fertility of bull spermatozoa frozen in whole milk extender with trehalose, taurine, or blood serum. J. Dairy. Sci. 76:1908-1913. https://doi.org/10.3168/jds.S0022-0302(93)77524-4
  21. Foote, R. H., Brockett, C. C. and Kaproth, M. T. 2002. Motility and fertility of bull sperm in whole milk extender containing antioxidants. Anim. Reprod. Sci. 71:13-23. https://doi.org/10.1016/S0378-4320(02)00018-0
  22. Fraser, L. R., Abeydeera, L. R. and Niwa, K. 1995. $Ca^{2+}$-regulating mechanisms that modulate bull sperm capacitation and acrosome alexocytosis as determined by chlortetracycline analysis. Mol. Reprod. Dev. 40:233-241. https://doi.org/10.1002/mrd.1080400213
  23. Funahashi, H. and Sano, T. 2005. Select antioxidants improve the function of extended boar semen stored at 10${^{\circ}C}$. Theriogenology 63:1605-1616. https://doi.org/10.1016/j.theriogenology.2004.06.016
  24. Gillan, L., Evans, G. and Maxwell, W. M. 1997. Capacitation status and fertility of fresh and frozen-thawed ram spermatozoa. Reprod. Fertil. Dev. 9:481-487. https://doi.org/10.1071/R96046
  25. Harrison, R. A. P., Ashworth, P. J. C. and Miller, N. G. A. 1996. Bicarbonate/CO2, an effector of capacitation, induces a rapid and reversible change in the lipid architecture of boar sperm plasma membranes. Mol. Reprod. Dev. 45:378-391. https://doi.org/10.1002/(SICI)1098-2795(199611)45:3<378::AID-MRD16>3.0.CO;2-V
  26. Hu, J. H., Li, Q. W., Jiang, Z. L., Yang, H., Zhang, S. S. and Zhao, H. W. 2009. The cryoprotective effect of trehalose supplementation on boar spermatozoa quality. Reprod. Domest. Anim. 44:571-575. https://doi.org/10.1111/j.1439-0531.2007.00975.x
  27. Irvine, D. S. 1996. Glutathione as a treatment for male infertility. Rev. Reprod. 1:6-12. https://doi.org/10.1530/ror.0.0010006
  28. Jeyendran, R. S., Van der Ven. H. H., Prez-Pelaez, M., Carbo, B. G. and Zaneveld, L. J. 1984. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil. 70:219-228. https://doi.org/10.1530/jrf.0.0700219
  29. de Lamirande, E. and Gagnon, C. 1992. Reactive oxygen species and human spermatozoa. II. Depletion of adenosine triphosphate plays an important role in the inhibition of sperm motility. J. Androl. 13:379-386.
  30. Liu, J. L., Kusakabe, H., Chang, C. C., Suzuki, H., Schmidt, D. W., Julian, M., Pfeffer, R., Bormann, C. L., Tian, X. C., Yanagimachi, R. and Yang, X. 2004. Freeze-dried sperm fertilization leads to full-term development in rabbits. Biol. Reprod. 70:1776-1781. https://doi.org/10.1095/biolreprod.103.025957
  31. Liu, Z., Foote, R. H. and Brockett, C. C. 1998. Survival of bull sperm frozen at different rates in media varying in osmolarity. Cryobiology 37:219-330. https://doi.org/10.1006/cryo.1998.2117
  32. Martins-Bessa, A., Rocha, A. and Mayenco-Aguirre, A. 2009. Effects of taurine and hypotaurine supplementation and ionophore concentrations on post-thaw acrosome reaction of dog spermatozoa. Theriogenology 71:248-253. https://doi.org/10.1016/j.theriogenology.2008.07.006
  33. Maxwell, W. M. C. and Watson, P. F. 1996. Recent progress in the preservation of ram semen. Anim. Reprod. Sci. 42:55-65. https://doi.org/10.1016/0378-4320(96)01544-8
  34. Medeiros, C. M., Forell, F., Oliveira, A. T. and Rodrigues, J. L. 2002. Current status of sperm cryopreservation: why isn't it better? Theriogenology 57:327-344. https://doi.org/10.1016/S0093-691X(01)00674-4
  35. Meizel, S., Lui, C. W., Working, P. K. and Mrsny, R. J. 1980. Taurine and hypotaurine: their effects on motility, capacitation and the acrosome reaction of hamster sperm in vitro and their presence in sperm and reproductive track fluids of several mammals. Dev. Growth Differ. 22:483-494. https://doi.org/10.1111/j.1440-169X.1980.00483.x
  36. Michael, A., Alexopoulos, C., Pontiki, E., Hadjipavlou-Litina, D., Saratsis, P. and Boscos, C. 2007. Effect of antioxidant supplementation on semen quality and reactive oxygen species of frozen-thawed canine spermatozoa. Theriogenology 68:204-212. https://doi.org/10.1016/j.theriogenology.2007.04.053
  37. Molinia, F. C., Evans, G., Quintana Casares, P. I. and Maxwell, W. M. C. 1994. Effect of monosaccharides and disaccharides in Tris-based diluents on motility, acrosome integrity and fertility of pellet frozen ram spermatozoa. Anim. Reprod. Sci. 36:113-122. https://doi.org/10.1016/0378-4320(94)90058-2
  38. Oh, S. A., You, Y. A., Park, Y. J. and Pang, M. G. 2010. The sperm penetration assay predicts the litter size in pigs. Int. J. Androl. 33:604-612.
  39. Pena, A. I., Lopez-Lugilde, L., Barrio, M., Becerra, J. J., Quintela, L. A. and Herradon, P. G. 2003. Studies on the intracellular Ca2+ concentration of thawed dog spermatozoa: influence of Equex from different sources, two thawing diluents and post-thaw incubation in capacitating conditions. Reprod. Domest. Anim. 38:27-35. https://doi.org/10.1046/j.1439-0531.2003.00391.x
  40. Purdy, P. H. 2006. A review on goat sperm cryopreservation. Small Rum. Res. 6:215-225.
  41. Sariozkan, S., Bucak, M. N., Tuncer, P. B., Ulutas, P. A. and Bilgen, A. 2009. The influence of cysteine and taurine on microscopicoxidative stress parameters and fertilizing ability of bull semen following cryopreservation. Cryobiology 58:134-138. https://doi.org/10.1016/j.cryobiol.2008.11.006
  42. Shiva Shankar Reddy, N., Jagan Mohanarao, G. and Atreja, S. K. 2010. Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation. Anim. Reprod. Sci. 119:183-190. https://doi.org/10.1016/j.anireprosci.2010.01.012
  43. Storey, B. T. 1997. Biochemistry of the induction and prevention of lipoperoxidative damage in human spermatozoa. Mol. Hum. Reprod. 3:203-213. https://doi.org/10.1093/molehr/3.3.203
  44. Storey, B. T., Noiles, E. E. and Thompson, K. A. 1998. Comparison of glycerol, other polyols, trehalose, and raffinose to provide a defined cryoprotectant medium for mouse sperm cryopreservation. Cryobiology 37:46-58. https://doi.org/10.1006/cryo.1998.2097
  45. Szasz, F., Sirivaidyapong, S. M., Cheng, F. P., Voorhout, W. F., Marks, A., Colenbrander, B., Solti, L. and Gadella, B. M. 2000. Detection of calcium ionophore induced membrane changes in dog sperm as a simple method to predict the cryopreservability of dog semen. Mol. Reprod. Dev. 55:289-298. https://doi.org/10.1002/(SICI)1098-2795(200003)55:3<289::AID-MRD7>3.0.CO;2-K
  46. Upreti, G. C., Jensen, K., Munday, R., Duganzich, D. M., Vishwanath, R. and Smith, J. F. 1998. Studies on aromatic amino acid oxidase activity in ram spermatozoa: role of pyruvate as an antioxidant. Anim. Reprod. Sci. 51:275-287. https://doi.org/10.1016/S0378-4320(98)00082-7
  47. Uysal, O., Bucak, M. N., Yavas, I. and Varush, O. 2007. Effect of various antioxidants on the quaility of frozen-thawed bull semen. J. Anim. Vet. Adv. 6:1362-1366.
  48. Yanagimach, R. 1994. Mammarian fertilization. In: Knobil, E. (Ed.). The physiology of Reproduction. Raven Press, NewYolk, U. S. A. PP. 189-317.

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