Comparison of Viability, ATP and In vitro Fertilization of Boar Sperm Stored at 4℃ in the Three Different Diluents

DOI QR코드

DOI QR Code

Yi, Y.J.;Li, Z.H.;Kim, E.S.;Song, E.S.;Cong, P.Q.;Zhang, Y.H.;Lee, S.H.;Lee, J.W.;Park, Chang-Sik

  • 투고 : 2008.01.21
  • 심사 : 2008.03.26
  • 발행 : 2008.08.01

초록

This study was designed to develop a method of liquid storage of boar sperm at $4^{\circ}C$ by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N-acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at $4^{\circ}C$ for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at $4^{\circ}C$. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to $20{\times}10^5$ sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of $2.5{\times}10^5$ sperm/ml was lower than in concentrations of 5, 10 and $20{\times}10^5$5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at $2.5{\times}10^5/ml$ sperm concentration was significantly lower than at 5, 10 and $20{\times}10^5sperm/ml$ concentrations. In conclusion, GPL4 diluent can be stored at $4^{\circ}C$ for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes.

키워드

Fertilization;Liquid Semen;Oocyte;Porcine

참고문헌

  1. Reed, H. C. B. 1985. Current use of frozen boar semen: Future need of frozen boar semen. Proc. First Intern. Conf. on Deep Freezing of Boar Semen, Uppsala, Sweden, pp. 225-237.
  2. Selles, J., J. Gadea, R. Romar, C. Matas and S. Ruiz. 2003. Analysis of in vitro fertilizing capacity to evaluate the freezing procedures of boar semen and to predict the subsequent fertility. Reprod. Dom. Anim. 38:66-72. https://doi.org/10.1046/j.1439-0531.2003.00406.x
  3. Visser, D. and S. Salamon. 1974. Effect of composition of trisbased diluents on survival of boar spermatozoa following deep-freezing. Aust. J. Biol. Sci. 27:485-497. https://doi.org/10.1071/BI9740485
  4. Waberski, D., K. F. Weitze, D. Rath and H. P. Sallmann. 1989. Effect of bovine serum albumin and zwitterionic buffers on stored liquid boar semen. Zuchthygiene 24:128-133. https://doi.org/10.1111/j.1439-0531.1989.tb00430.x
  5. Nagai, T., K. Niwa and A. Iritani. 1984. Effect of sperm concentration during preincubation in a defined medium on fertilization in vitro of pig follicular oocytes. J. Reprod. Fertil. 70:271-275. https://doi.org/10.1530/jrf.0.0700271
  6. Paquignon, M. 1985. Freezing and thawing extenders for boar spermatozoa. Proc. First Intern. Conf. on Deep Freezing of Boar Semen, Uppsala, Sweden, pp. 129-145.
  7. Petzoldt, R. and H. Nehring. 1988. Grundlagen der Biologie und Konservierung von Saugerspermien. Biol. Rundschau. 26:79-89.
  8. Pursel, V. G. and L. A. Johnson. 1975. Freezing of boar spermatozoa: Fertilizing capacity with concentrated semen and a new thawing procedure. J. Anim. Sci. 40:99-102. https://doi.org/10.2527/jas1975.40199x
  9. Harrison, R. A. P., H. M. Dott and G. C. Foster. 1982. Bovine serum albumin, sperm motility, and the dilution effect. J. Exp. Zool. 222:81-88. https://doi.org/10.1002/jez.1402220111
  10. Hu, J. H., Q. W. Li, G. Li, X. Y. Chen, H. Yang, S. S. Zhang and L. Q. Wang. 2006. The cryoprotective effect on frozen-thawed boar semen of egg yolk low density lipoproteins. Asian-Aust. J. Anim. Sci. 19(4):486-494. https://doi.org/10.5713/ajas.2006.486
  11. Lehninger, A. L. 1975. Biochemistry Lehninger. Worth Publishers Inc., New York, NK, 10016, p. 266
  12. Hamano, S. and Y. Toyoda. 1986. In vitro fertilization of pig eggs with ejaculated spermatozoa preincubated at high sperm concentration. Jpn. J. Anim. Reprod. 32:177-183. https://doi.org/10.1262/jrd1977.32.177
  13. Abeydeera, L. R. and B. N. Day. 1997. Fertilization and subsequent development in vitro of pig oocytes inseminated in a modified tris-buffered medium with frozen-thawed ejaculated spermatozoa. Biol. Reprod. 57:729-734. https://doi.org/10.1095/biolreprod57.4.729
  14. Buhr, M. M. 1990. Preservation of boar sperm alters membrane molecular dynamics. Reprod. Domest. Anim. (Suppl 1), pp. 81-93.
  15. Courtens, J. L. and M. Paquignon. 1985. Ultrastructure of fresh, frozen and frozen-thawed spermatozoa of the boar. Proc. First Intern. Conf. on Deep Freezing of Boar Semen, Uppsala, Sweden, pp. 61-87.
  16. Coy, P., S. Ruiz, R. Romar, I. Campos and J. Gadea. 1999. Maturation, fertilization and complete development of porcine oocytes matured under different systems. Theriogenol. 5:799-812.
  17. Ford, S. R. and F. R. Leach. 1998. Bioluminescent assay of the adenylate energy charge. Methods Mol. Biol. 102:69-81.
  18. White, I. G. 1993. Lipids and Calcium uptake of sperm in relation to cold shock and preservation: a review. Reprod. Fertil. Dev. 5:639-658. https://doi.org/10.1071/RD9930639
  19. Yi, Y. J., Y. A. Kwon, H. J. Ko and C. S. Park. 2002. Effects of diluent component, freezing rate, thawing time and thawing temperature on acrosome morphology and motility of frozenthawed boar sperm. Asian-Aust. J. Anim. Sci. 15(11):1553-1558. https://doi.org/10.5713/ajas.2002.1553
  20. Waberski, D., S. Meding, G. Dirksaen, K. F. Weitze, C. Lewiding and R. Hahn. 1994. Fertility of long term-stored boar semen: influence of extender (Androhep and Kiev), storage time and plasma droplets in the semen. Anim. Reprod. Sci. 36:145-151. https://doi.org/10.1016/0378-4320(94)90061-2
  21. Wang, W. H., K. Niwa and K. Okuda. 1991. In vitro penetration of pig oocytes matured in culture by frozen-thawed ejaculated spermatozoa. J. Reprod. Fertil. 93:491-496. https://doi.org/10.1530/jrf.0.0930491
  22. Weitze, K. F. 1991. Long-term storage of extended boar semen. Proc. 2nd Int. Conf. on Boar Semen preservation II, Beltsville, Maryland, USA, pp. 231-253
  23. Westendorf, P., L. Richter and H. Treu. 1975. Zur Tiefgefrierung von Ebersperma. Labor-und Besamungsergebnisse mit dem Hulsenberger Pailetten-Verfahren. Dtsch. Tieraztl. Wschr. 82:261-267.
  24. Cong, P. Q., E. S. Song, E. S. Kim, Y. J. Yi and C. S. Park. 2007. Effects of BSA, PVA, gonadotropins and follicle shell on in vitro maturation and in vitro fertilization of porcine oocytes. Reprod. Dev. Biol. 31(2):61-69.
  25. Yi, Y. J., Z. H. Li, E. S. Kim, E. S. Song, H. B. Kim, P. Q. Cong, J. M. Lee and C. S. Park. 2008. Comparison of motility and normal acrosome of boar sperm with or without cold shock resistance in liquid semen at 17$^{\circ}$C and 4$^{\circ}$C, and frozen-thawed semen. Asian-Aust. J. Anim. Sci. 21(2):190-197. https://doi.org/10.5713/ajas.2008.70351