Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Antioxidant Effect of Alpha-Linolenic Acid during In Vitro Maturation in Porcine Oocytes

  • Lee, Ji-Eun (College of Animal Life Sciences, Kangwon National University) ;
  • Hwangbo, Yong (College of Animal Life Sciences, Kangwon National University) ;
  • Kim, Hwa-Young (College of Animal Life Sciences, Kangwon National University) ;
  • Cheong, Hee-Tae (College of Veterinary Medicine, Kangwon National University) ;
  • Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University) ;
  • Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
  • Received : 2017.11.21
  • Accepted : 20171131
  • Published : 2017.11.30
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Abstract

Alpha-linolenic acid (ALA) is one of n-3 polyunsaturated fatty acids and found mainly in the chloroplasts. Many studies have been reported that intracellular reactive oxygen species (ROS) in mammalian oocytes were reduced by supplementation of ALA in in vitro maturation (IVM) medium. Based on these reports, we expected that ALA acts as an antioxidant during IVM of porcine oocytes. Therefore, the objective of this study was to investigate the antioxidant effect of ALA supplementation during IVM in porcine oocytes. The cumulus-oocyte complexes (COCs) were incubated in IVM medium containing $200{\mu}m$ $H_2O_2$ or $H_2O_2$ with $50{\mu}m$ ALA for 44 h. Nuclear maturation stage of oocytes was evaluated using aceto-orcein method. For measurement of oxidative stress state, intracellular ROS and glutathione (GSH) levels were measured using carboxy-DCFDA and cell tracker red, respectively. In results, oocytes in metaphase-II (MII) stage development was significantly reduced in $H_2O_2$ group compared to non-treated control group $61.84{\pm}1.42%$ and 80.00%, respectively; p<0.05) and it was slightly recovered by treatment of ALA ($69.76{\pm}1.67%$; p<0.05). The intracellular GSH levels was decreased in $H_2O_2$ groups compared with control groups, but it was enhanced by ALA treatment (p<0.05). On the contrary, $H_2O_2$ treatment increased intracellular ROS level in oocytes and $H_2O_2$-induced ROS was decreased by treatment of ALA (p<0.05). Our findings suggested that ALA treatment under oxidative stress condition improve oocyte maturation via elevated GSH and reduced ROS levels in oocytes. Therefore, these results suggest that ALA have an antioxidative ability and it could be used as antioxidant in in vitro production system of porcine embryo.

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References

  1. Alvarez GM, Morado SA, Soto MP, Dalvit GC, Cetica PD (2015): The control of reactive oxygen species influences porcine oocyte in vitro maturation. Reprod Dom Anim 50:200-205. https://doi.org/10.1111/rda.12469
  2. Bagg MA, Nottle MB, Armstrong DT, Grupen CG (2007): Relationship between follicle size and oocyte developmental competence in prepubertal and adult pigs. Reprod Fertil Dev 19:797-803. https://doi.org/10.1071/RD07018
  3. Flores D, Souza V, Betancourt M, Teteltitla M, Gonzalez-Marquez H, Casas E, Bonilla E, Ramirez-Noguera P, Gutierrez-Ruiz MC, Ducolomb (2017): Oxidative stress as a damage mechanism in porcine cumulus-oocyte complexes exposed to malathion during in vitro maturation. Environ Toxicol 32(6):1669-1678. https://doi.org/10.1002/tox.22384
  4. Guerin P, El Mouatassim S, Menezo Y (2001). Oxidative stress and protection against reactive oxygen species in the pre-implantation embryo and its surroundings. Hum Reprod Update 7(2):175-189. https://doi.org/10.1093/humupd/7.2.175
  5. Hu J, Cheng D, Gao X, Bao J, Ma X, Wang H (2012). Vitamin C enhances the in vitro development of porcine pre-implantation embryos by reducing oxidative stress. Reprod Dom Anim 47(6):873-879. https://doi.org/10.1111/j.1439-0531.2011.01982.x
  6. Jeon Y, Yoon JD, Cai L, Hwang SU, Kim E, Zheng Z, Lee E, Kim DY, Hyun SH (2014): Supplementation of zinc on oocyte in vitro maturation improves preimplatation embryonic development in pigs. Theriogenology 82:866-874. https://doi.org/10.1016/j.theriogenology.2014.06.021
  7. Kere M, Siriboon C, Lo NW, Nguyen NT, Ju JC (2013): Ascorbic acid improves the developmental competence of porcine oocytes after parthenogenetic activation and somatic cell nuclear transplantation. J Reprod Dev 59:78-84.
  8. Kim J, You J, Hyun SH, Lee G, Lim J, Lee E (2010): Developmental competence of morphologically poor oocytes in relation to follicular size and oocyte diameter in the pig. Mol Reprod Dev 77:330-339.
  9. Kitagawa Y, Suzuki K, Yoneda A, Watanabe T (2004): Effects of oxygen concentration and antioxidants on the in vitro developmental ability, production of reactive oxygen species, and DNA fragmentation in porcine embryos. Theriogenology 62:1186-1197. https://doi.org/10.1016/j.theriogenology.2004.01.011
  10. Kowaltowski AJ, Vercesi AE (1999): Mitochondrial damage induced by conditions of oxidative stress. Free Radic Biol Med 26:463-71. https://doi.org/10.1016/S0891-5849(98)00216-0
  11. Kwak SS, Cheong SA, Joen Y, Lee E, Choi KC, Jeung EB, Hyun SH (2012): The effects o resveratrol on porcine oocyte in vitro maturation and subsequent embryonic development after parthenogenetic activation and in vitro fertilization. Theriogenology 78:86-101. https://doi.org/10.1016/j.theriogenology.2012.01.024
  12. Lee JE, Hwangbo Y, Kim HY, Lee WH, Cheong HT, Yang BK, Park CK (2017): Effect of alpha-linolenic acid on oocyte maturation and embryo development in pigs. Dev & Reprod 21:205. https://doi.org/10.12717/DR.2017.21.2.205
  13. Lee S, Jin JX, Khoirinaya C, Kim GA, Lee BC (2016a): Lanosterol influences cytoplasmic maturation of pig oocytes in vitro and improves preimplantation development of cloned embryos. Theriogenology 85:575-584. https://doi.org/10.1016/j.theriogenology.2015.09.041
  14. Lee Y, Lee H, Park B, Elahi J, Lee J, Lee ST, Park CK, Hyun SH, Lee E (2016b): Alpha-linolenic acid treatment during oocyte maturation enhances embryonic development by influencing mitogen-activated protein kinase activity and intraoocyte glutathione content in pigs. J Anim Sci 94:3255-3263. https://doi.org/10.2527/jas.2016-0384
  15. Li Y, Nhang ZZ, He CJ, Zhu KF, Xu ZY, Ma T, Tao JL, Liu GS (2015): Melatonin protects porcine oocyte in vitro maturation from heat stress. J Pineal Res 59:365-375. https://doi.org/10.1111/jpi.12268
  16. Luberda Z (2005): The role of glutathione in mammalian gametes. Reprod Biol 5:5-17.
  17. Marei WF, Wathes D, Fouladi-Nashta AA (2012): Differential effects of linoleic and alpha-linolenic fatty acids on spatial and temporal mitochondrial distribution and activity in bovine oocytes. Reprod Fertil Dev 24:679-690. https://doi.org/10.1071/RD11204
  18. Marei WF, Wathes DC, Fouladi-Nashta AA (2009): The effect of linolenic acid on bovine oocyte maturation and development. Biol Reprod 81:1064-1072. https://doi.org/10.1095/biolreprod.109.076851
  19. Marei WF, Wathes DC, Fouladi-Nashta AA (2010) Impact of linoleic acid on bovine oocyte maturation and embryo development. Reproduction 139:979-988. https://doi.org/10.1530/REP-09-0503
  20. Matsunaga R, Funahashi H (2016): Supplementation with cumulus cell masses improves the in vitro meiotic competence of porcine cumulus-oocytes complexes derived from small follicles. Reprod Domest Anim 52:672-679.
  21. McKeegan PJ, Sturmey RG (2011): The role of fatty acids in oocyte and early embryo development. Reprod Fertil Dev 24:59-67.
  22. Meister A, Anderson ME (1983): Glutathione. Ann Rev Biochem 52:711-760. https://doi.org/10.1146/annurev.bi.52.070183.003431
  23. Mishra A, Reddy I, Gupta P, Mondal S (2016). Lcarnitine mediated reduction in oxidative stress and alteration in transcript level of antioxidant enzymes in sheep embryos produced in vitro. Reprod Dom Anim 51:311-321. https://doi.org/10.1111/rda.12682
  24. Natarajan R, Shankar MB, Munuswamy D (2010). Effect of ${\alpha}$-tocopherol supplementation on in vitro maturation of sheep oocytes and in vitro development of preimplantation sheep embryos to the blastocyst stage. J Assist Reprod Genet 27(8):483-490. https://doi.org/10.1007/s10815-010-9430-7
  25. Pompella A, Visvikis A, Paolicchi A, De Tata V, Casini AF (2003): The changing faces od glutathione, a cellular protagonist. Biochem Pharmacol 66(8):1499-1503. https://doi.org/10.1016/S0006-2952(03)00504-5
  26. Rocha-Frigoni, NA, Leao BC, Dall'Acqua PC, Mingoti GZ (2016): Improving the cytoplasmic maturation of bovine oocytes matured in vitro with intracellular and/or extracellular antioxidants is not associated with increased rates of embryo development. Theriogenology 86:1897-1905. https://doi.org/10.1016/j.theriogenology.2016.06.009
  27. Sovernigo TC, Adona PR, Monzani PS, Guemra S, Barros FDA, Lopes FG, Leal CLV (2017): Effects of supplementation of medium with different antioxidants during in vitro maturation of bovine oocytes on subsequent embryo production. Reprod Dom Anim 52:561-569. https://doi.org/10.1111/rda.12946
  28. Sturmey RG, Reis A, Leese HJ, McEvoy TG (2009): Role of fatty acids in energy provision during oocyte maturation and early embryo development. Reprod Dom Anim 44(Suppl 3):50-58. https://doi.org/10.1111/j.1439-0531.2009.01402.x
  29. Taweechaipaisankul A, Jin JX, Lee S, Kim GA, Lee BC (2016): The effects of canthaxanthin on porcine oocyte maturation and embryo development in vitro after parthenogenetic activation and somatic cell nuclear transfer. Reprod Dom Anim 51:870-876. https://doi.org/10.1111/rda.12748
  30. Wu GQ, Jia BY, Li JJ, Fu XW, Zhou GB, Hou YP, Zhu SE (2011): L-carnitine enhances oocyte maturation and development of parthenogenetic embryos in pigs. Theriogenology 76:785-793. https://doi.org/10.1016/j.theriogenology.2011.04.011
  31. Yazaki T, Hiradate Y, Hoshino Y, Tanemura K, Sato E (2013): L-carnitine improves hydrogen peroxideinduced impairment of nuclear maturation in porcine oocytes. Anim Sci J 84:395-402. https://doi.org/10.1111/asj.12016
  32. Zhang X, Wu XQ, Lu S, Guo YL, Ma X (2006): Deficit of mitochondria-derived ATP during oxidative stress impairs mouse MII oocyte spindles. Cell research 16(10):841-850. https://doi.org/10.1038/sj.cr.7310095