Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Effect of the addition of 4OHE2 and quercetin in culture media on ROS levels and gene expression in mouse blastocysts

Nuria Hernández https://orcid.org/0000-0002-6812-1189 A B * , Soledad Sánchez-Mateos B , Marta López-Morató B C , Francisco M. Sánchez-Margallo B and Ignacio S. Álvarez D E *
+ Author Affiliations
- Author Affiliations

A Criopreservation Facility, Biomedicine Institute, Seville, Spain.

B Assisted Reproduction Unit, Jesús Usón Minimally Invasive Surgery Centre, Cáceres, Spain.á

C Genetics Unit, Hospital HLA Vistahermosa, Alicante, Spain.

D Department of Cell Biology, University of Extremadura, Badajoz, Spain.

E Instituto Extremeño de Reproduccion Asistida (IERA), Badajoz, Spain.


Handling Editor: Ye Yuan

Reproduction, Fertility and Development 34(15) 980-990 https://doi.org/10.1071/RD22041
Published online: 9 September 2022

© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Aims: The main objective of this work is to elucidate whether Quercetin (Qc) and 4-Hidroxistradiol (4OHE2) decrease the level of reactive oxygen species (ROS) in in vitro obtained embryos and to analyse which genes are activated under the treatments that could explain this improvement.

Methods: Oxidative stress was induced during embryo culture by H2O2 treatment and ROS production was measured and compared with embryos treated with Qc or 4OHE2. Gene expression was analysed by Q-PCR in control embryos obtained in utero (IU) or by IVF and compared with the levels found in embryos cultured with Qc or 4OHE2 to determine the effect of these compounds.

Key results: Qc strongly reduces ROS levels in embryos after a treatment of 4 h. On the contrary, 4OHE2 had no effect in reducing ROS levels in embryos. The addition of these molecules to the culture media upregulate several hypoxia-related genes when Qc is added to the culture media, and implantation-related genes when 4OHE2 is used.

Conclusions: Qc is a very strong antioxidant molecule that when used for short periods of time during culture can reduce ROS levels and improve embryo quality by activating antioxidant enzymes. 4OHE2 supplementation, despite having no effects in reducing ROS levels, acts directly in the molecular signalling implicated in the implantation process and could be also considered as a supplement for embryo culture during IVF.

Implications: Proper supplementation of the culture media could greatly improve the quality of embryos cultured in vitro, resulting in better results in IVF clinics.

Keywords: 4-Hidroxistradiol, blastocysts, embryo development, hypoxia, implantation, mouse, oxidative stress, quercetin, ROS.


References

Abdelrazik, H, Sharma, R, Mahfouz, R, and Agarwal, A (2009). L-Carnitine decreases DNA damage and improves the in vitro blastocyst development rate in mouse embryos. Fertility and Sterility 91, 589–596.
L-Carnitine decreases DNA damage and improves the in vitro blastocyst development rate in mouse embryos.Crossref | GoogleScholarGoogle Scholar |

Agarwal, A, Aponte-Mellado, A, Premkumar, BJ, Shaman, A, and Gupta, S (2012). The effects of oxidative stress on female reproduction: a review. Reproductive Biology and Endocrinology 10, 49.
The effects of oxidative stress on female reproduction: a review.Crossref | GoogleScholarGoogle Scholar |

Aherne, SA, and O’Brien, NM (2000). Mechanism of protection by the flavonoids, quercetin and rutin, against tert-butylhydroperoxide- and menadione-induced DNA single strand breaks in Caco-2 cells. Free Radical Biology and Medicine 29, 507–514.
Mechanism of protection by the flavonoids, quercetin and rutin, against tert-butylhydroperoxide- and menadione-induced DNA single strand breaks in Caco-2 cells.Crossref | GoogleScholarGoogle Scholar |

Balasubramanian, S, Son, WJ, Kumar, BM, et al. (2007). Expression pattern of oxygen and stress-responsive gene transcripts at various developmental stages of in vitro and in vivo preimplantation bovine embryos. Theriogenology 68, 265–275.
Expression pattern of oxygen and stress-responsive gene transcripts at various developmental stages of in vitro and in vivo preimplantation bovine embryos.Crossref | GoogleScholarGoogle Scholar |

Bogacz, A, Mikołajczak, PŁ, Wolek, M, et al. (2021). Combined effects of methyldopa and flavonoids on the expression of selected factors related to inflammatory processes and vascular diseases in human placenta cells – an in vitro study. Molecules 26, 1259.
Combined effects of methyldopa and flavonoids on the expression of selected factors related to inflammatory processes and vascular diseases in human placenta cells – an in vitro study.Crossref | GoogleScholarGoogle Scholar |

Bolouki, A, Zal, F, and Alaee, S (2020). Ameliorative effects of quercetin on the preimplantation embryos development in diabetic pregnant mice. The Journal of Obstetrics and Gynaecology Research 46, 736–744.
Ameliorative effects of quercetin on the preimplantation embryos development in diabetic pregnant mice.Crossref | GoogleScholarGoogle Scholar |

Boots, AW, Haenen, GRMM, and Bast, A (2008). Health effects of quercetin: from antioxidant to nutraceutical. European Journal of Pharmacology 585, 325–337.
Health effects of quercetin: from antioxidant to nutraceutical.Crossref | GoogleScholarGoogle Scholar |

Caramelo, C, Peña Deudero, JJ, Castilla, A, Justo, S, De Solis, AJ, Neria, F, Peñate, S, and Gonzalez-Pacheco, FR (2006). Respuesta a la hipoxia. un mecanismo sistémico basado en el control de la expresión génica. Medicina 66, 155–164.

Carroll, VA, and Ashcroft, M (2006). Role of hypoxia-inducible factor (HIF)-1α versus HIF-2α in the regulation of HIF target genes in response to hypoxia, insulin-like growth factor-I, or loss of von hippel-lindau function: implications for targeting the HIF pathway. Cancer Research 66, 6264–6270.
Role of hypoxia-inducible factor (HIF)-1α versus HIF-2α in the regulation of HIF target genes in response to hypoxia, insulin-like growth factor-I, or loss of von hippel-lindau function: implications for targeting the HIF pathway.Crossref | GoogleScholarGoogle Scholar |

Chronopoulou, E, and Harper, JC (2015). IVF culture media: past, present and future. Human Reproduction Update 21, 39–55.
IVF culture media: past, present and future.Crossref | GoogleScholarGoogle Scholar |

De Simoni, S, Linard, D, Hermans, E, Knoops, B, and Goemaere, J (2013). Mitochondrial peroxiredoxin-5 as potential modulator of mitochondria-ER crosstalk in MPP+−induced cell death. Journal of Neurochemistry 125, 473–485.
Mitochondrial peroxiredoxin-5 as potential modulator of mitochondria-ER crosstalk in MPP+−induced cell death.Crossref | GoogleScholarGoogle Scholar |

Dong, GZ, Kim, CY, Cho, WS, Shin, YJ, and Lee, K (2006). Cryoprotective role of the flavonoid quercetin in mammalian cells. Korean Journal of Genetics 28, 269–277.

Dunwoodie, SL (2009). The role of hypoxia in development of the mammalian embryo. Developmental Cell 17, 755–773.
The role of hypoxia in development of the mammalian embryo.Crossref | GoogleScholarGoogle Scholar |

el Hajj, N, and Haaf, T (2013). Epigenetic disturbances in in vitro cultured gametes and embryos: implications for human assisted reproduction. Fertility and Sterility 99, 632–641.
Epigenetic disturbances in in vitro cultured gametes and embryos: implications for human assisted reproduction.Crossref | GoogleScholarGoogle Scholar |

Ferrali, M, Signorini, C, Caciotti, B, Sugherini, L, Ciccoli, L, Giachetti, D, and Comporti, M (1997). Protection against oxidative damage of erythrocyte membrane by the flavonoid quercetin and its relation to iron chelating activity. FEBS Letters 416, 123–129.
Protection against oxidative damage of erythrocyte membrane by the flavonoid quercetin and its relation to iron chelating activity.Crossref | GoogleScholarGoogle Scholar |

Ganguly, A, McKnight, RA, Raychaudhuri, S, Shin, B-C, Ma, Z, Moley, K, and Devaskar, SU (2007). Glucose transporter isoform-3 mutations cause early pregnancy loss and fetal growth restriction. American Journal of Physiology - Endocrinology and Metabolism 292, E1241–E1255.
Glucose transporter isoform-3 mutations cause early pregnancy loss and fetal growth restriction.Crossref | GoogleScholarGoogle Scholar |

Gao, N, Nester, RA, and Sarkar, MA (2004). 4-Hydroxy estradiol but not 2-hydroxy estradiol induces expression of hypoxia-inducible factor 1α and vascular endothelial growth factor A through phosphatidylinositol 3-kinase/Akt/FRAP pathway in OVCAR-3 and A2780-CP70 human ovarian carcinoma cells. Toxicology and Applied Pharmacology 196, 124–135.
4-Hydroxy estradiol but not 2-hydroxy estradiol induces expression of hypoxia-inducible factor 1α and vascular endothelial growth factor A through phosphatidylinositol 3-kinase/Akt/FRAP pathway in OVCAR-3 and A2780-CP70 human ovarian carcinoma cells.Crossref | GoogleScholarGoogle Scholar |

Gardner, DK, and Wale, PL (2013). Analysis of metabolism to select viable human embryos for transfer. Fertility and Sterility 99, 1062–1072.
Analysis of metabolism to select viable human embryos for transfer.Crossref | GoogleScholarGoogle Scholar |

Harvey, AJ (2007). The role of oxygen in ruminant preimplantation embryo development and metabolism. Animal Reproduction Science 98, 113–128.
The role of oxygen in ruminant preimplantation embryo development and metabolism.Crossref | GoogleScholarGoogle Scholar |

Harvey, AJ, Kind, KL, and Thompson, JG (2002). REDOX regulation of early embryo development. Reproduction 123, 479–486.
REDOX regulation of early embryo development.Crossref | GoogleScholarGoogle Scholar |

Heijnen, CGM, Haenen, GRMM, Minou Oostveen, R, Stalpers, EM, and Bast, A (2002). Protection of flavonoids against lipid peroxidation: the structure activity relationship revisited. Free Radical Research 36, 575–581.
Protection of flavonoids against lipid peroxidation: the structure activity relationship revisited.Crossref | GoogleScholarGoogle Scholar |

Hernández, N, López-Morató, M, Perianes, MJ, Sánchez-Mateos, S, Casas-Rua, V, Domínguez-Arroyo, JA, Sánchez-Margallo, FM, and Álvarez, IS (2021). 4-Hydroxyestradiol improves mouse embryo quality, epidermal growth factor-binding capability in vitro and implantation rates. Molecular Human Reproduction 27, gaaa075.
4-Hydroxyestradiol improves mouse embryo quality, epidermal growth factor-binding capability in vitro and implantation rates.Crossref | GoogleScholarGoogle Scholar |

Huang, YT, Hwang, J-J, Lee, P-P, Ke, F-C, Huang, J-H, Huang, C-J, Kandaswami, C, Middleton, E, and Lee, M-T (1999). Effects of luteolin and quercetin, inhibitors of tyrosine kinase, on cell growth and metastasis-associated properties in A431 cells overexpressing epidermal growth factor receptor. British Journal of Pharmacology 128, 999–1010.
Effects of luteolin and quercetin, inhibitors of tyrosine kinase, on cell growth and metastasis-associated properties in A431 cells overexpressing epidermal growth factor receptor.Crossref | GoogleScholarGoogle Scholar |

Jeong, W, Bazer, FW, Song, G, and Kim, J (2016). Expression of hypoxia-inducible factor-1 by trophectoderm cells in response to hypoxia and epidermal growth factor. Biochemical and Biophysical Research Communications 469, 176–182.
Expression of hypoxia-inducible factor-1 by trophectoderm cells in response to hypoxia and epidermal growth factor.Crossref | GoogleScholarGoogle Scholar |

Kaneuchi, M, Sasaki, M, Tanaka, Y, Sakuragi, N, Fujimoto, S, and Dahiya, R (2003). Quercetin regulates growth of Ishikawa cells through the suppression of EGF and cyclin D1. International Journal of Oncology 22, 159–164.
Quercetin regulates growth of Ishikawa cells through the suppression of EGF and cyclin D1.Crossref | GoogleScholarGoogle Scholar |

Kang, J-T, Moon, JH, Choi, J-Y, et al. (2016). Effect of antioxidant flavonoids (quercetin and taxifolin) on in vitro maturation of porcine oocytes. Asian-Australasian Journal of Animal Sciences 29, 352–358.
Effect of antioxidant flavonoids (quercetin and taxifolin) on in vitro maturation of porcine oocytes.Crossref | GoogleScholarGoogle Scholar |

Karagenc, L, Sertkaya, Z, Ciray, N, Ulug, U, and Bahçeci, M (2004). Impact of oxygen concentration on embryonic development of mouse zygotes. Reproductive BioMedicine Online 9, 409–417.
Impact of oxygen concentration on embryonic development of mouse zygotes.Crossref | GoogleScholarGoogle Scholar |

Kazuo, U, Taijiro, M, Takashi, S, Tadashi, H, Machiko, T, Yoichi, K, and Shozo, T (1977). In vitro transformation of hamster embryo cells by quercetin. Toxicology Letters 1, 175–178.
In vitro transformation of hamster embryo cells by quercetin.Crossref | GoogleScholarGoogle Scholar |

Khanduja, KL, Verma, A, and Bhardwaj, A (2001). Impairment of human sperm motility and viability by quercetin is independent of lipid peroxidation. Andrologia 33, 277–281.
Impairment of human sperm motility and viability by quercetin is independent of lipid peroxidation.Crossref | GoogleScholarGoogle Scholar |

Kind, KL, Collett, RA, Harvey, AJ, and Thompson, JG (2005). Oxygen-regulated expression of GLUT-1, GLUT-3, and VEGF in the mouse blastocyst. Molecular Reproduction and Development 70, 37–44.
Oxygen-regulated expression of GLUT-1, GLUT-3, and VEGF in the mouse blastocyst.Crossref | GoogleScholarGoogle Scholar |

Kirby, RS (2018). Assisted reproductive technology and developmental outcomes. Pediatrics 142, e20183072.
Assisted reproductive technology and developmental outcomes.Crossref | GoogleScholarGoogle Scholar |

Krussel, JS, Behr, B, Milki, AA, et al. (2001). Vascular endothelial growth factor (VEGF) mRNA splice variants are differentially expressed in human blastocysts. Molecular Human Reproduction 7, 57–63.
Vascular endothelial growth factor (VEGF) mRNA splice variants are differentially expressed in human blastocysts.Crossref | GoogleScholarGoogle Scholar |

Lee, SK, Oh, KH, Chung, AY, et al. (2015). Protective role of quercetin against cisplatin-induced hair cell damage in zebrafish embryos. Human & Experimental Toxicology 34, 1043–1052.
Protective role of quercetin against cisplatin-induced hair cell damage in zebrafish embryos.Crossref | GoogleScholarGoogle Scholar |

Li, XX, Lee, K-B, Lee, JH, Kim, KJ, Kim, EY, Han, K-W, Park, K-S, Yu, J, and Kim, MK (2014). Glutathione and cysteine enhance porcine preimplantation embryo development in vitro after intracytoplasmic sperm injection. Theriogenology 81, 309–314.
Glutathione and cysteine enhance porcine preimplantation embryo development in vitro after intracytoplasmic sperm injection.Crossref | GoogleScholarGoogle Scholar |

Lim, H, Paria, BC, Das, SK, et al. (1997). Multiple female reproductive failures in cyclooxygenase 2-deficient mice. Cell 91, 197–208.
Multiple female reproductive failures in cyclooxygenase 2-deficient mice.Crossref | GoogleScholarGoogle Scholar |

Lu, H, Forbes, RA, and Verma, A (2002). Hypoxia-inducible factor 1 activation by aerobic glycolysis implicates the Warburg effect in carcinogenesis. Journal of Biological Chemistry 277, 23111–23115.
Hypoxia-inducible factor 1 activation by aerobic glycolysis implicates the Warburg effect in carcinogenesis.Crossref | GoogleScholarGoogle Scholar |

Ma, Y-Y, Chen, H-W, and Tzeng, C-R (2017). Low oxygen tension increases mitochondrial membrane potential and enhances expression of antioxidant genes and implantation protein of mouse blastocyst cultured in vitro. Journal of Ovarian Research 10, 47.
Low oxygen tension increases mitochondrial membrane potential and enhances expression of antioxidant genes and implantation protein of mouse blastocyst cultured in vitro.Crossref | GoogleScholarGoogle Scholar |

Mantikou, E, Bontekoe, S, van Wely, M, Seshadri, S, Repping, S, and Mastenbroek, S (2013a). Low oxygen concentrations for embryo culture in assisted reproductive technologies. Human Reproduction Update 19, 209.
Low oxygen concentrations for embryo culture in assisted reproductive technologies.Crossref | GoogleScholarGoogle Scholar |

Mantikou, E, Youssef, MAFM, van Wely, M, van der Veen, F, Al-Inany, HG, Repping, S, and Mastenbroek, S (2013b). Embryo culture media and IVF/ICSI success rates: a systematic review. Human Reproduction Update 19, 210–220.
Embryo culture media and IVF/ICSI success rates: a systematic review.Crossref | GoogleScholarGoogle Scholar |

Marions, L, and Danielsson, kG (1999). Expression of cyclo-oxygenase in human endometrium during the implantation period. Molecular Human Reproduction 5, 961–965.
Expression of cyclo-oxygenase in human endometrium during the implantation period.Crossref | GoogleScholarGoogle Scholar |

Mlcek, J, Jurikova, T, Skrovankova, S, and Sochor, J (2016). Quercetin and its anti-allergic immune response. Molecules 21, 623.
Quercetin and its anti-allergic immune response.Crossref | GoogleScholarGoogle Scholar |

Nagao, A, Seki, M, and Kobayashi, H (1999). Inhibition of xanthine oxidase by flavonoids. Bioscience, Biotechnology, and Biochemistry 63, 1787–1790.
Inhibition of xanthine oxidase by flavonoids.Crossref | GoogleScholarGoogle Scholar |

Nelissen, ECM, Van Montfoort, APA, Smits, LJM, Menheere, PPCA, Evers, JLH, Coonen, E, Derhaag, JG, Peeters, LL, Coumans, AB, and Dumoulin, JCM (2013). IVF culture medium affects human intrauterine growth as early as the second trimester of pregnancy. Human Reproduction 28, 2067–2074.
IVF culture medium affects human intrauterine growth as early as the second trimester of pregnancy.Crossref | GoogleScholarGoogle Scholar |

Neumann, CA, Cao, J, and Manevich, Y (2009). Peroxiredoxin 1 and its role in cell signaling. Cell Cycle 8, 4072–4078.
Peroxiredoxin 1 and its role in cell signaling.Crossref | GoogleScholarGoogle Scholar |

Paria, BC, Das, SK, and Dey, SK (1997). Embryo implantation requires estrogendirected uterine preparation and catecholestrogen-mediated embyronic activation. Advances in Pharmacology 42, 840–843.
Embryo implantation requires estrogendirected uterine preparation and catecholestrogen-mediated embyronic activation.Crossref | GoogleScholarGoogle Scholar |

Pringle, KG, Kind, KL, Sferruzzi-Perri, AN, Thompson, JG, and Roberts, CT (2010). Beyond oxygen: complex regulation and activity of hypoxia inducible factors in pregnancy. Human Reproduction Update 16, 415–431.
Beyond oxygen: complex regulation and activity of hypoxia inducible factors in pregnancy.Crossref | GoogleScholarGoogle Scholar |

Rabbani, ML, and Rogers, PA (2001). Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats. Reproduction 122, 85–90.
Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats.Crossref | GoogleScholarGoogle Scholar |

Rakhit M, Gokul SR, Agarwal A, du Plessis SS (2013) Antioxidant strategies to overcome OS in IVF-Embryo transfer. In ‘Studies on women’s health.’ pp. 237–262. (Humana Press).
| Crossref |

Rempel, MA, Hester, B, DeHaro, H, Hong, H, Wang, Y, and Schlenk, D (2009). Effects of 17β-estradiol, and its metabolite, 4-hydroxyestradiol on fertilization, embryo development and oxidative DNA damage in sand dollar (Dendraster excentricus) sperm. Science of The Total Environment 407, 2209–2215.
Effects of 17β-estradiol, and its metabolite, 4-hydroxyestradiol on fertilization, embryo development and oxidative DNA damage in sand dollar (Dendraster excentricus) sperm.Crossref | GoogleScholarGoogle Scholar |

Rizzo, A, Roscino, MT, Binetti, F, and Sciorsci, RL (2012). Roles of reactive oxygen species in female reproduction. Reproduction in Domestic Animals 47, 344–352.
Roles of reactive oxygen species in female reproduction.Crossref | GoogleScholarGoogle Scholar |

Salama, SA, Kamel, M, Awad, M, Ben Nasser, A-HB, Al-Hendy, A, Botting, S, and Arrastia, C (2008). Catecholestrogens induce oxidative stress and malignant transformation in human endometrial glandular cells: protective effect of catechol-O-methyltransferase. International Journal of Cancer 123, 1246–1254.
Catecholestrogens induce oxidative stress and malignant transformation in human endometrial glandular cells: protective effect of catechol-O-methyltransferase.Crossref | GoogleScholarGoogle Scholar |

Schmidt, S, Hommel, A, Gawlik, V, et al. (2009). Essential role of glucose transporter GLUT3 for post-implantation embryonic development. Journal of Endocrinology 200, 23–33.
Essential role of glucose transporter GLUT3 for post-implantation embryonic development.Crossref | GoogleScholarGoogle Scholar |

Semenza, GL (2000). HIF-1: mediator of physiological and pathophysiological responses to hypoxia. Journal of Applied Physiology 88, 1474–1480.
HIF-1: mediator of physiological and pathophysiological responses to hypoxia.Crossref | GoogleScholarGoogle Scholar |

Sistani Karampour, N, Arzi, A, Najafzadeh Varzi, H, Mohammadian, B, and Rezaei, M (2014). Quercetin preventive effects on theophylline-induced anomalies in rat embryo. Jundishapur Journal of Natural Pharmaceutical Products 9, 17834.
Quercetin preventive effects on theophylline-induced anomalies in rat embryo.Crossref | GoogleScholarGoogle Scholar |

Takahashi, M (2012). Oxidative stress and redox regulation on in vitro development of mammalian embryos. Journal of Reproduction and Development 58, 1–9.
Oxidative stress and redox regulation on in vitro development of mammalian embryos.Crossref | GoogleScholarGoogle Scholar |

Takeuchi, M, Seki, M, Furukawa, E, Takahashi, A, Saito, K, Kobayashi, M, Ezoe, K, Fukui, E, Yoshizawa, M, and Matsumoto, H (2017). Improvement of implantation potential in mouse blastocysts derived from IVF by combined treatment with prolactin, epidermal growth factor and 4-hydroxyestradiol. Molecular Human Reproduction 23, 557–570.
Improvement of implantation potential in mouse blastocysts derived from IVF by combined treatment with prolactin, epidermal growth factor and 4-hydroxyestradiol.Crossref | GoogleScholarGoogle Scholar |

Truong, T, and Gardner, DK (2017). Antioxidants improve IVF outcome and subsequent embryo development in the mouse. Human Reproduction 32, 2404–2413.
Antioxidants improve IVF outcome and subsequent embryo development in the mouse.Crossref | GoogleScholarGoogle Scholar |

Truong, TT, Soh, YM, and Gardner, DK (2016). Antioxidants improve mouse preimplantation embryo development and viability. Human Reproduction 31, 1445–1454.
Antioxidants improve mouse preimplantation embryo development and viability.Crossref | GoogleScholarGoogle Scholar |

Tsutomu, N, Munetaka, Y, Toshihiko, O, and Shunro, K (1993). Suppression of active oxygen-induced cytotoricity by flavonoids. Biochemical Pharmacology 45, 265–267.
Suppression of active oxygen-induced cytotoricity by flavonoids.Crossref | GoogleScholarGoogle Scholar |

Ullah, O, Li, Z, Ali, I, Xu, L, Liu, H, Shah, SZA, and Fang, N (2019). Pterostilbene alleviates hydrogen peroxide-induced oxidative stress via nuclear factor erythroid 2 like 2 pathway in mouse preimplantation embryos. Journal of Reproduction and Development 65, 73–81.
Pterostilbene alleviates hydrogen peroxide-induced oxidative stress via nuclear factor erythroid 2 like 2 pathway in mouse preimplantation embryos.Crossref | GoogleScholarGoogle Scholar |

Wale, PL, and Gardner, DK (2012). Oxygen regulates amino acid turnover and carbohydrate uptake during the preimplantation period of mouse embryo development. Biology of Reproduction 87, 24.
Oxygen regulates amino acid turnover and carbohydrate uptake during the preimplantation period of mouse embryo development.Crossref | GoogleScholarGoogle Scholar |

Wale, PL, and Gardner, DK (2016). The effects of chemical and physical factors on mammalian embryo culture and their importance for the practice of assisted human reproduction. Human Reproduction Update 22, 2–22.
The effects of chemical and physical factors on mammalian embryo culture and their importance for the practice of assisted human reproduction.Crossref | GoogleScholarGoogle Scholar |

Wang, GL, Jiang, BH, Rue, EA, and Semenza, GL (1995). Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proceedings of the National Academy of Sciences of the United States of America 92, 5510–5514.
Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension.Crossref | GoogleScholarGoogle Scholar |

Wenger, RH (2002). Cellular adaptation to hypoxia: O2 sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression. The FASEB Journal 16, 1151–1162.
Cellular adaptation to hypoxia: O2 sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression.Crossref | GoogleScholarGoogle Scholar |

Yoon, J, Juhn, K-M, Ko, J-K, et al. (2013). Effects of oxygen tension and IGF-I on HIF-1α protein expression in mouse blastocysts. Journal of Assisted Reproduction and Genetics 30, 99–105.
Effects of oxygen tension and IGF-I on HIF-1α protein expression in mouse blastocysts.Crossref | GoogleScholarGoogle Scholar |

Yu, S, Long, H, Lyu, Q-F, Zhang, Q-H, Yan, Z-G, Liang, H-X, Chai, W-R, Yan, Z, Kuang, Y-P, and Qi, C (2014). Protective effect of quercetin on the development of preimplantation mouse embryos against hydrogen peroxide-induced oxidative injury. PLoS ONE 9, e89520.
Protective effect of quercetin on the development of preimplantation mouse embryos against hydrogen peroxide-induced oxidative injury.Crossref | GoogleScholarGoogle Scholar |