IMPROVEMENT OF SHEEP OOCYTE COMPETENCE IN VITRO THROUGH GLUTATHIONE ADDITION
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Abstract
Oocyte competence is a primary factor for the success of embryo production in vitro. This research aims to determine the effect of supplementation glutathione in sheep oocytes’ maturation and fertilization rates in vitro. In the first research, maturation oocytes were conducted in a medium supplemented with GSH at graded doses for 24 hours. In the second research, 1.0 mM GSH (the best dose from research I) was supplemented in the maturation medium, fertilization medium, and their combination for 14 hours. The higher oocyte maturation rates were significant (P < 0.05) in the GSH supplementation of 1.0 mM (87.00%). Furthermore, supplementation of GSH on the maturation medium resulted in a higher normal fertilization rate (56.43%) than in other groups. It is concluded that GSH supplementation on the maturation medium is more effective in increasing the competence of sheep oocytes to be mature and results in improved fertilization rates.
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References
Adeoye O, Olawumi J, Opeyemi A, Christiania O (2018) Review on the role of glutathione on oxidative stress and infertility. J Bras Reprod Assist 22:61–66. https://doi.org/10.5935/1518-0557.20180003
Bardaweel SK, Gul M, Alzweiri M, Ishaqat A, Alsalamat HA, Bashatwah RM (2018) Reactive oxygen species: The dual role in physiological and pathological conditions of the human body. Eurasian J Med 50:193–201. https://doi.org/10.5152/eurasianjmed.2018.17397
Chapman JC, Michael SD (2003) Proposed mechanism for sperm chromatin condensation/decondensation in the male rat. Reprod Biol Endocrinol 1:1–7
Fan H-Y, Sun Q-Y (2019) Oocyte Meiotic Maturation. The Ovary 181–203. https://doi.org/10.1016/b978-0-12-813209-8.00012-1
Forman HJ, Zhang H, Rinna A (2009) Glutathione: Overview of its protective roles, measurement, and biosynthesis. Mol Aspects Med 30:1–12. https://doi.org/10.1016/j.mam.2008.08.006.Glutathione
Gatimel N, Moreau J, Parinaud J, Léandri RD (2020) Need for choosing the ideal pH value for IVF culture media. J Assist Reprod Genet 37:1019–1028. https://doi.org/10.1007/s10815-020-01726-5
Gulo FDK, Karja NWK, Setiadi MA (2020) Cysteamine in maturation medium enhances nuclear maturation and fertilization rate of sheep oocytes in vitro. HAYATI J Biosci 27:290–295. https://doi.org/10.4308/hjb.27.4.290
Itahashi T, Oikawa T, Numabe T (2022) Effects of glutathione treatments during sperm washing and in vitro fertilization on the in vitro early development of embryos of Japanese Black cattle. J Reprod Dev 68:74–78. https://doi.org/https://doi.org/10.1262/jrd.2021-070
Jiang Y, He Y, Pan X, Wang P, Yuan X, Ma B (2023) Advances in Oocyte Maturation In Vivo and In Vitro in Mammals. Int J Mol Sci 24:9059. https://doi.org/10.3390/ijms24109059
Jomova K, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, Valko M (2024) Several lines of antioxidant defense against oxidative stress: antioxidant enzymes, nanomaterials with multiple enzyme-mimicking activities, and low-molecular-weight antioxidants. Springer Berlin Heidelberg
Kemper JM, Liu Y, Afnan M, Mol BWJ, Morbeck DE (2023) What happens to abnormally fertilized embryos? A scoping review. Reprod Biomed Online 46:802–807. https://doi.org/10.1016/j.rbmo.2023.02.005
Kozlov A V, Javadov S, Sommer N (2024) Cellular ROS and Antioxidants : Physiological and Pathological Role. Antioxidants 13:1–12. https://doi.org/https://doi.org/10.3390/antiox13050602
Lennicke C, Cocheme HM (2021) Review Redox metabolism : ROS as specific molecular regulators of cell signaling and function. Mol Cell 81:3691–3707. https://doi.org/10.1016/j.molcel.2021.08.018
Li F (2018) Exogenous glutathione improves intracellular glutathione synthesis via the γ ‐ glutamyl cycle in bovine zygotes and cleavage embryos. J Cell Physiol 234:1–11. https://doi.org/10.1002/jcp.27497
Liu N, Si X, Ji Y, Yang Q, Bai J, He Y, Jia H, Wu Z (2023) L -Proline improves the cytoplasmic maturation of mouse oocyte by regulating glutathione-related redox homeostasis. Theriogenology 195:159–167. https://doi.org/10.1016/j.theriogenology.2022.10.023
Lu SC (2013) Glutathione synthesis. Biochim Biophys Acta - Gen Subj 1830:3143–3153. https://doi.org/10.1016/j.bbagen.2012.09.008.GLUTATHIONE
Luberda Z (2005) The role of glutathione in mammalian gametes. Reprod Biol 5:5–17
Menchaca A, Santos-neto PC, Cuadro F, Souza-neves M, Crispo M (2018) From reproductive technologies to genome editing in small ruminants : an embryo ’ s journey. Anim Reprod 15:984. https://doi.org/10.21451/1984-3143-AR2018-0022
Moulavi F, Hosseini SM (2018) Diverse patterns of cumulus cell expansion during in vitro maturation reveal heterogeneous cellular and molecular features of oocyte competence in dromedary camel. Theriogenology 119:259–267. https://doi.org/10.1016/j.theriogenology.2018.07.010
Nugroho AP, Supriatna I, Setiadi MA (2017) Penambahan Glutathione pada Medium Fertilisasi Efektif Mendukung Pembentukan Pronukleus dan Perkembangan Awal Embrio Sapi (Supplementation of Glutathione in Fertilization Medium Effectively Support Normal Pronucleus Formation and Early Bovine Embryonic Development Rate. J Vet 18:327. https://doi.org/10.19087/jveteriner.2017.18.3.327
Qu J, Hu H, Niu H, Sun X, Li Y (2023) Melatonin restores the declining maturation quality and early embryonic development of oocytes in aged mice. Theriogenology 210:110–118. https://doi.org/10.1016/j.theriogenology.2023.07.021
Rahmatullah R, Setiadi MA, Supriatna I (2022) Heparin and Hypotaurine Supplementation Improve the Fertilization Rate of Sheep Oocytes Matured in Media Containing L-Carnitine in Vitro. J Kedokt Hewan - Indones J Vet Sci 16:121–126. https://doi.org/10.21157/j.ked.hewan.v16i4.27339
Rakha SI, Elmetwally MA, Ali HE, Balboula A, Mahmoud AM, Zaabel SM (2022) Importance of Antioxidant Supplementation during In Vitro Maturation of Mammalian Oocytes. Vet Sci 9:439. https://doi.org/https://doi.org/10.3390/vetsci9080439
Reczek CR, Chandel NS (2016) ROS-dependent signal transduction. Curr Opin Cell Biol 33:8–13. https://doi.org/10.1016/j.ceb.2014.09.010.
Ren J, Hao Y, Liu Z, Li S, Wang C, Wang B, Liu Y, Liu G, Dai Y (2021) Effect of exogenous glutathione supplementation on the in vitro developmental competence of ovine oocytes. Theriogenology 173:144–155. https://doi.org/10.1016/j.theriogenology.2021.07.025
Sciorio R, Smith G (2019) Embryo culture at a reduced oxygen concentration of 5 %: a mini review. Zygote 26:1–7. https://doi.org/10.1017/S0967199419000522
Shi W, Qin C, Yang Y, Yang X, Fang Y, Zhang B, Wang D, Feng W, Shi D (2025) Urolithin A Protects Porcine Oocytes from Artificially Induced Oxidative Stress Damage to Enhance Oocyte Maturation and Subsequent Embryo Development. Int J Mol Sci 26:3037. https://doi.org/https://doi.org/10.3390/ijms26073037
Shirazi A, Golestanfar A, Bashiri M, Ahmadi E (2018) Male Pronuclear Formation and Embryo Development Following Intracytoplasmic Injection of Ovine Pretreated Sperm. Avicenna J Med Biotechnol 10:41–48
Souza-Fabjan JMG, Leal GR, Ana C, Monteiro S, Ivan R, Pereira T, Barbosa NO, José V, Freitas F (2023) In vitro embryo production in small ruminants : what is still missing ? Anim Reprod 20:1–24. https://doi.org/https://doi.org/10.1590/1984-3143-AR2023-0055
Sun W, Pang Y, Liu Y, Hao H, Zhao X, Qin T, Zhu H, Du W (2015) Exogenous glutathione supplementation in culture medium improves the bovine embryo development after in vitro fertilization. Theriogenology 84:716–723. https://doi.org/10.1016/j.theriogenology.2015.05.001
Suzuki K, Eriksson B, Shimizu H, Nagai T (2000) Effect of hyaluronan on monospermic penetration of porcine oocytes fertilized in vitro. Int J Androl 23:13–21. https://doi.org/https://doi.org/10.1046/j.1365-2605.2000.t01-1-00198.x
Takeo T, Nakagata N (2011) Reduced Glutathione Enhances Fertility of Frozen / Thawed C57BL / 6 Mouse Sperm after Exposure to Methyl-Beta-Cyclodextrin 1. Biol Reprod 85:1066–1072. https://doi.org/10.1095/biolreprod.111.092536
Takeshima T, Usui K, Mori K, Asai T, Yasuda K, Kuroda S, Yumura Y (2021) Oxidative stress and male infertility. Reprod Med Biol 20:41–52. https://doi.org/10.1002/rmb2.12353
Ufer C, Wang CC (2011) The roles of glutathione peroxidases during embryo development. Front Mol Neurosci 4:1–15. https://doi.org/10.3389/fnmol.2011.00012
Unnikrishnan V, Kastelic J, Thundathil J (2021) Intracytoplasmic sperm injection in cattle. Genes (Basel) 12:1–18. https://doi.org/10.3390/genes12020198
Villalpando-Rodriguez GE, Gibson SB (2021) Reactive Oxygen Species (ROS) Regulates Different Types of Cell Death by Acting as a Rheostat. Oxid Med Cell Longev 2021:1–17. https://doi.org/10.1155/2021/9912436
Wang Y, Wang Q, Ji Q, An P (2025) Supplementation with N-Acetyl-L-cysteine during in vitro maturation improves goat oocyte developmental competence by regulating oxidative stress. Theriogenology 235:221–230. https://doi.org/10.1016/j.theriogenology.2025.01.016
Wei X, Manandhar L, Kim H, Chhetri A, Hwang J, Jang G, Park C, Park R (2025) Pexophagy and Oxidative Stress: Focus on Peroxisomal Proteins and Reactive Oxygen Species (ROS) Signaling Pathways. Antioxidants 14:126. https://doi.org/10.3390/antiox14020126
Yang S, Lian G (2020) ROS and diseases: role in metabolism and energy supply. Mol Cell Biochem 467:1–12. https://doi.org/10.1007/s11010-019-03667-9
Ye ZW, Zhang J, Townsend DM, Tew KD (2015) Oxidative stress, redox regulation and diseases of cellular differentiation. Biochim Biophys Acta 1850:1607–1621. https://doi.org/10.1016/j.bbagen.2014.11.010
Yusuf AM (2024) Promise of In vitro Embryo Production Technology for Improvement of Cattle Reproductive Potential. EAS J Vet Med Sci 6:44–58. https://doi.org/10.36349/easjvms.2024.v06i02.003
