Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Effect of Monothioglycerol on ROS Inhibition, Mitochondrial Activity, and DNA Integrity in Frozen-thawed Miniature Pig Sperm

Monothiolglycerol이 동결 융해 후 미니돼지 정자의 활성산소 억제, 미토콘드리아 활성 그리고 DNA Integrity에 미치는 영향

  • Park, Soo-Jung (Department of Biological Science, Bio-Nano Technology, Gachon University) ;
  • Kim, Dae-Young (Department of Biological Science, Bio-Nano Technology, Gachon University)
  • 박수정 (가천대학교 바이오나노대학 생명과학과) ;
  • 김대영 (가천대학교 바이오나노대학 생명과학과)
  • Received : 2013.09.09
  • Accepted : 2013.09.24
  • Published : 2013.09.30
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Abstract

Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

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References

  1. Aitken RJ, Jones KT and Robertson SA. 2012. Reactive oxygen species and sperm function in sickness and in health. J. Androl. 33: 1096-1106. https://doi.org/10.2164/jandrol.112.016535
  2. Chatterjee S and Gagnon C. 2001. Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol. Reprod. Dev. 59: 451-458. https://doi.org/10.1002/mrd.1052
  3. Cheema RS, Bansal AK and Bilaspuri GS. 2009. Manganese provides antioxidant protection for sperm cryopreservation that may offer new consideration for clinical fertility. Oxid. Med. Cell. Longev. 2: 152-159. https://doi.org/10.4161/oxim.2.3.8804
  4. Cormier N, Sirard MA and Bailey JL. 1997. Premature capacitation of bovine spermatozoa is initiated by cryopreservation. J. Androl. 18: 461-468.
  5. Critser JK and Mobraaten LE. 2000. Cryopreservation of murine spermatozoa. Ilar. J. 41: 197-206. https://doi.org/10.1093/ilar.41.4.197
  6. Gray JE, Starmer J, Lin VS, Dickinson BC and Magnuson T. 2013. Mitochondrial hydrogen peroxide and defective cholesterol efflux prevent in vitro fertilization by cryopreserved inbred mouse sperm. Biol. Reprod. 89: 17. https://doi.org/10.1095/biolreprod.113.109157
  7. Gutierrez-Perez O, Juarez-Mosqueda Mde L, Carvajal SU and Ortega ME. 2009. Boar spermatozoa cryopreservation in low glycerol/trehalose enriched freezing media improves cellular integrity. Cryobiology 58: 287-292. https://doi.org/10.1016/j.cryobiol.2009.02.003
  8. Hammerstedt RH, Graham JK and Nolan JP. 1990. Cryopreservation of mammalian sperm: what we ask them to survive. J. Androl. 11: 73-88.
  9. Hu JH, Li QW, Jiang ZL, Yang H, Zhang SS and Zhao HW. 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
  10. Hwang YJ and Kim DY. 2013. Effects of ginsenoside-Rg1 on post-thawed miniature pig sperm motility, mitochondria activity, and membrane integrity. J. Emb. Trans. 28: 63-71. https://doi.org/10.12750/JET.2013.28.1.63
  11. Jha PK, Paul AK, Rahman MB, Tanjin M, Bari FY and Alam MGS. 2013. Improvement of preservation quality of chilled bull semen using $\alpha$-tocopherol as an antioxidant. J. Emb. Trans. 28: 31-39. https://doi.org/10.12750/JET.2013.28.1.31
  12. Kaneko T, Yamamura A, Ide Y, Ogi M, Yanagita T and Nakagata N. 2006. Long-term cryopreservation of mouse sperm. Theriogenology 66: 1098-1101. https://doi.org/10.1016/j.theriogenology.2006.02.049
  13. Lee SY, Ahn JH, Ko KW, Kim J, Jeong SW, Kim IK and Kim HS. 2010. Prostaglandin A2 activates intrinsic apoptotic pathway by direct interaction with mitochondria in HL-60 cells. Prostaglandins Other Lipid. Mediat. 91: 30-37. https://doi.org/10.1016/j.prostaglandins.2009.12.003
  14. Ostermeier GC, Wiles MV, Farley JS and Taft RA. 2008. Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation. PLoS One. 3: e2792. https://doi.org/10.1371/journal.pone.0002792
  15. Park CH, Kim SW, Hwang YJ and Kim DY. 2012. Cryopreservation with trehalose reduced sperm shromatin damage in miniature pig. J. Emb. Trans. 27: 107-111.
  16. Rosato MP and Iaffaldano N. 2013. Cryopreservation of rabbit semen: comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification. Theriogenology 79: 508-516. https://doi.org/10.1016/j.theriogenology.2012.11.008
  17. Seltzer WK, Dhariwal G, McKelvey HA and McCabe ER. 1986. 1-Thioglycerol: inhibitor of glycerol kinase activity in vitro and in situ. Life Sci. 39: 1417-1424. https://doi.org/10.1016/0024-3205(86)90545-X
  18. Sontakke SD, Umapathy G, Sivaram V, Kholkute SD and Shivaji S. 2004. Semen characteristics, cryopreservation, and successful artificial insemination in the Blue Rock pigeon (Columba livia). Theriogenology 62: 139-153. https://doi.org/10.1016/j.theriogenology.2003.08.018
  19. Takahashi H and Liu C. 2010. Archiving and distributing mouse lines by sperm cryopreservation, IVF, and embryo transfer. Methods Enzymol. 476: 53-69. https://doi.org/10.1016/S0076-6879(10)76004-3
  20. Tsuneto M, Yamazaki H, Yoshino M, Yamada T and Hayashi S. 2005. Ascorbic acid promotes osteoclastogenesis from embryonic stem cells. Biochem. Biophys. Res. Commun. 335: 1239-1246. https://doi.org/10.1016/j.bbrc.2005.08.016
  21. Watson PF. 2000. The causes of reduced fertility with cryopreserved semen. Anim. Reprod. Sci. 60-61: 481-492. https://doi.org/10.1016/S0378-4320(00)00099-3
  22. Yamaguchi T, Takamura H, Matoba T and Terao J. 1998. HPLC method for evaluation of the free radical-scavenging activity of foods by using 1,1-diphenyl-2-picrylhydrazyl. Biosci. Biotechnol. Biochem. 62: 1201-1204. https://doi.org/10.1271/bbb.62.1201
  23. Zou Y, Tao TZ, Tian Y, Zhu JL, Cao LJ, Deng XM and Li JB. 2013. Ginsenoside Rg1 improves survival in a murine model of polymicrobial sepsis by suppressing the inflammatory response and apoptosis of lymphocytes. J. Surg. Res. 183: 760-766. https://doi.org/10.1016/j.jss.2013.01.068