3,4-Dihydroxyflavone Acts as an Antioxidant and Antiapoptotic Agent to Support Bovine Embryo Development In Vitro

Lee, Keum Sil; Kim, Eun Young; Jeon, Kilsoo; Cho, Ssang-Goo; Han, Young Joon; Yang, Byoung‐Chul; Lee, Sungsoo; Ko, Mon Suck; Riu, Key Jung; Lee, Hoon Taek; Park, Se Pill

doi:10.1262/jrd.10-029a

Cited by 22 publications

(18 citation statements)

References 27 publications

(25 reference statements)

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“…Generally, it is considered that stimulated levels of the enzyme have evolved to address increased free-radical production during inflammatory episodes. The high MnSOD expression levels found in our results were consistent with those in previous reports that suggested that increased levels of MnSOD might be related to inadequate reprogramming of the donor genome in NT embryos [34][35][36]. However, the high expression levels of MnSOD in Group 3 might have been due to these embryos' increased vitality.…”

Section: Discussionsupporting

confidence: 91%

Effects of Repetitive Ionomycin Treatment on In Vitro Development of Bovine Somatic Cell Nuclear Transfer Embryos

Kim¹,

Lee²,

Jeong³

et al. 2012

Journal of Reproduction and Development

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Abstract. To artificially activate embryos in somatic cell nuclear transfer (SCNT), chemical treatment with ionomycin has been used to induce transient levels of Ca 2+ and initiate reprogramming of embryos. Ca 2+ oscillation occurs naturally several times after fertilization (several times with 15-to 30-min intervals). This indicates how essential additional Ca 2+ influx is for successful reprogramming of embryos. Hence, in this report, the experimental design was aimed at improving the developmental efficiency of cloned embryos by repetitive Ca 2+ transients rather than the commonly used ionomycin treatment (4 min). To determine optimal Ca 2+ inflow conditions, we performed three different repetitive ionomycin (10 μM) treatments in reconstructed embryos: Group 1 (4-min ionomycin treatment, once), Group 2 (30-sec treatment, 4 times, 15-min intervals) and Group 3 (1-min treatment, 4 times, 15-min intervals). Pronuclear formation rates were checked to assess the effects of repetitive ionomycin treatment on reprogramming of cloned embryos. Cleavage rates were investigated on day 2, and the formation rates of blastocysts (BLs) were examined on day 7 to demonstrate the positive effect of repeated ionomycin treatment. In Group 3, a significant increase in BL formation was observed [47/200 (23.50%), 44/197 (22.33%) and 69/195 (35.38%) in Groups 1, 2 and 3, respectively]. Culturing embryos with different ionomycin treatments caused no significant difference among the groups in terms of the total cell number of BLs (164.3, 158.5 and 145.1, respectively). Additionally, expression of the anti-apoptotic Bcl-2 gene and MnSOD increased significantly in Group 3, whereas the expression of the pro-apoptotic Bax decreased statistically. In conclusion, the present study demonstrated that repeated ionomycin treatment is an improved activation method that can increase the developmental competence of SCNT embryos by decreasing the incidence of apoptosis. Key words: Apoptosis, Bovine embryos, Ca 2+ oscillation, SCNT (J. Reprod. Dev. 58: [132][133][134][135][136][137][138][139] 2012) C a 2+ oscillation is a universal signaling cue that activates numerous cellular responses. Ca 2+ signaling is ubiquitous in some somatic cells and germ lines. In particular, in early embryo development, the fluctuating Ca 2+ concentration is a factor that brings about serial developmental mechanisms [1]. After fertilization, matured oocytes arrested in the metaphase stage of the second meiotic division (MII) turn into zygotes with subsequent signaling events such as depolarization, Ca 2+ oscillation, cortical reaction and pronucleus formation [2,3]. The exit from MII resumption and the further processes for forthcoming development do not occur before the concentration of intracellular Ca 2+ is increased by sperm penetration. Sperm is the initial signal for Ca 2+ release mediated by IP 3 (inositol trisphosphate), protein tyrosine kinases and PLC gamma (PLC-ζ in mammals), followed by reorganization of Ca 2+ -releasing machinery to trigger subsequent s...

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Section: Discussionsupporting

confidence: 91%

Effects of Repetitive Ionomycin Treatment on In Vitro Development of Bovine Somatic Cell Nuclear Transfer Embryos

Kim¹,

Lee²,

Jeong³

et al. 2012

Journal of Reproduction and Development

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“…Also supplementation of polyphenols promotes development of pig [122] and mouse [123] embryos as well as cow embryos [121,124]. Addition of flavonoids also affected the expressions of Mn-SOD, survivin, Bax-I, Glut-5 and IFN-τ genes in cow embryos as well as improving development [121].…”

Section: Development Of Embryosmentioning

confidence: 97%

“…They are characterized by the presence of several groups involved in phenolic structures and include the flavonoids and phenolic acids. Flavonoids promoted development of bovine embryos culture both in high (20%) and low (5%) O 2 tension with reducing the intracellular ROS [121]. Also supplementation of polyphenols promotes development of pig [122] and mouse [123] embryos as well as cow embryos [121,124].…”

Section: Development Of Embryosmentioning

confidence: 99%

Oxidative Stress and Redox Regulation on In Vitro Development of Mammalian Embryos

2012

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Abstract. Many factors affect development of mammalian preimplantation embryos in vitro. It is well known that in vitro development of bovine embryos is highly affected by culture condition including energy source, growth factors, pH or gas environment. Many efforts have been made towards the suitable environments which can successfully support embryo development in vitro. For a rapid growth and differentiation, embryo requires energy by utilizing ATP, NADPH with oxygen molecules. These energy substrates are produced from the electron transport chain in the mitochondria. In addition to energy production, reactive oxygen species (ROS) are also generated as by-product of such energy production system. ROS production is sensitively controlled by the balance of oxidizing and reducing status and affected by several antioxidant enzymes such as superoxide dismutase (SOD), Catalase, glutathione peroxidase (GPx) or low molecular weight thiols such as glutathione (GSH). Imbalance of oxidation and reduction causes production of excess ROS, which causes the developmental arrest, physical DNA damage, apoptosis induction or lipid peroxidation. Environmental oxygen condition during embryo culture also highly affects embryo development as well as intracellular redox balance. Several studies have revealed that regulation of intra-and extra-cellular reducing environment by reducing excess ROS by using antioxidants, reducing oxygen concentration are effective for improving embryo development. Also, recent studies have demonstrated the difference in gene expression affected by oxidative stress. This review briefly summarizes the effects of ROS and the role of redox balance on preimplantation embryos for improving the efficiency of in vitro production of mammalian embryos. Key words: Embryo development, Glutathione, Intracellular redox state, Reactive oxygen species (J. Reprod. Dev. 58: 1-9, 2012) H andling of oocytes and embryos in vitro is an effective technique for obtaining offspring by embryo transfer, investigating the mechanisms of early embryonic development and differentiation, gene functions by transgenic approach and research of somatic cell clones in mammals. To improve the pregnancy rates in in vitro produced (IVP) embryos, it is important to evaluate and improve the quality of embryos suitable for transfer. Therefore, increasing the number of good quality embryos as well as selection of superior quality embryos is one of the most practical and clinically useful approaches in embryo transfer of mammalian preimplantation embryos. It is well known that in vitro development of bovine embryos is highly affected by culture condition. Therefore, much effort has been made towards the effective culture methods that support embryo development in vitro. The detrimental effect of oxidative stress caused by a high oxygen concentration under culture condition on embryo development has been studied with the comparison of oxygen concentration in in vivo condition.It was found that embryo development is promoted under a low O 2 concent...

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“…It inhibits glutamate-triggered apoptosis induced by glutathione (GSH) depletion and ROS production and has antioxidant activity in neurons by acting as a selective tyrosine kinase receptor B agonist [27, 28]. In addition, 3,4-dihydroxyflavone (3,4-DHF) supports bovine embryo development in vitro as an antioxidant and anti-apoptotic agent [29], and 7,8-DHF appeared to have protective effect against oxidative stress [30]. However, the effects of 7,8-DHF for porcine oocytes and embryos have not been well investigated.…”

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confidence: 99%

Effect of 7,8-Dihydroxyflavone as an Antioxidant on In Vitro Maturation of Oocytes and Development of Parthenogenetic Embryos in Pigs

et al. 2013

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One of the factors that impairs in vitro produced porcine embryos is the oxidative stress that is mainly caused by the imbalance between reactive oxygen species (ROS) generation and antioxidants activity, especially that of glutathione (GSH). Here, we examined the effect of 7,8-dihydroxyflavone (7,8-DHF), a kind of flavonoid antioxidant, on porcine oocyte maturation and its developmental competence. Porcine oocytes were cultured in media supplemented with 0, 1, 5 and 10 μM 7,8-DHF during both in vitro maturation (IVM) and in vitro culture (IVC) after parthenogenetic activation. Maturation of oocytes was evaluated based on first polar body (PB) extrusion and intracellular GSH level, and developmental competence was assessed through observing cleavage and blastocyst formation. In each step, the levels of intracellular GSH and ROS were assessed by fluorescence intensity, and the apoptosis-related gene expression was examined using semiquantitative RT-PCR. The group treated with 1 μM 7,8-DHF during IVM and IVC showed increased cytoplasmic maturation and reached the blastocysts stage (36.1%) at a higher rate than the other groups (24.7, 16.0 and 10.3% for 0, 5 and 10 μM, P<0.05). In that group, the intracellular GSH level was significantly increased while ROS generation was significantly decreased after IVM and IVC (P<0.05). Moreover, it showed high expression of an anti-apoptotic gene (BCL2L1) and low expression of a pro-apoptotic gene (BAK1) (P<0.05). In conclusion, treatment with 1 μM 7,8-DHF during IVM and IVC showed an anti-apoptotic effect by increasing intracellular GSH synthesis and scavenging ROS and therefore improved the developmental competence of porcine embryos.

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3,4-Dihydroxyflavone Acts as an Antioxidant and Antiapoptotic Agent to Support Bovine Embryo Development In Vitro

Cited by 22 publications

References 27 publications

Effects of Repetitive Ionomycin Treatment on <i>In Vitro</i> Development of Bovine Somatic Cell Nuclear Transfer Embryos

Effects of Repetitive Ionomycin Treatment on <i>In Vitro</i> Development of Bovine Somatic Cell Nuclear Transfer Embryos

Oxidative Stress and Redox Regulation on <i>In Vitro</i> Development of Mammalian Embryos

Effect of 7,8-Dihydroxyflavone as an Antioxidant on <i>In Vitro</i> Maturation of Oocytes and Development of Parthenogenetic Embryos in Pigs

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