173 MELATONIN ALLEVIATES THE ENDOPLASMIC RETICULUM STRESS THROUGH THE REGULATING OF UNFOLDING PROTEIN RESPONSE SIGNALING DURING PORCINE OOCYTE MATURATION IN VITRO

2017 ◽  
Vol 29 (1) ◽  
pp. 195 ◽  
Author(s):  
J.-Y. Park ◽  
H.-J. Park ◽  
J.-W. Kim ◽  
S.-Y. Park ◽  
S.-G. Yang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Er Stress ◽  
Oocyte Maturation ◽  
Caspase 3 ◽  
Oxygen Species ◽  
Melatonin Treatment ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Protein Response

Unfolding protein response (UPR) is a defence mechanism during endoplasmic reticulum (ER) stress in mammalian cells. Especially, UPR genes and regulation of reactive oxygen species is involved in ER stress response on porcine oocyte maturation in vitro. Some studies have shown that melatonin treatment results in reducing oxidative stress, a protective function of free radical damage in oocyte maturation and embryo development. Also, melatonin has an important role in reducing reactive oxygen species and ER stress. However, it is unknown how the changes of UPR genes expression levels are affected the porcine oocyte maturation. In addition, there are no reports about ER stress recovery mechanism by melatonin during porcine oocyte maturation. Here, we investigated the UPR signal genes (Bip/Grp78, Atf4, p90/p50Atf6, and Xbp1) and ER-stress mediated apoptosis factors (Chop and Cleaved caspase 3) in porcine oocyte maturation in vitro. Expression of Chop and Cleaved caspase 3 mRNA levels were significantly increased (P < 0.01) in matured oocytes (metaphase II; 44 h) in vitro. Porcine oocytes were cultured in maturation medium with ER stress inducer, tunicamycin (Tm), and supplemented with various concentrations (1, 5, and 10 μg mL−1) of Tm for 0 to 44 h. Our results indicated that the proportion of matured oocytes was significantly decreased in Tm-treated groups in a dose-dependent manner (60.1 ± 1.3, 46.5 ± 2.1, and 38.9 ± 5.1% at 1, 5, and 10 μg mL−1 of Tm) compared with the control group (76.6 ± 1.9%). Likewise, mRNA expression of UPR regulator genes (Grp78/Bip, Aft4, Xbp1, Chop, and Cleaved caspase 3) was decreased by melatonin treatment (0.1 μM, 22–44 h) after pretreatment of Tm (5 μg mL−1, 0–22 h) during oocyte maturation. Our results demonstrated that the roles of melatonin as UPR signaling regulator for reducing ER stress are essential for promotion of porcine oocyte maturation and cumulus cell expansion of cumulus-oocyte complex. Moreover, the current study was initiated to confirm a functional link between effect of melatonin and regulating of UPR signaling in porcine oocytes maturation. These results suggest that melatonin improve the oocyte maturation and cumulus cells expansion by regulating of UPR signal genes against the ER stress during the porcine in vitro maturation process. This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01117604) and the Bio-industry Technology Development Program (316037–04–1-HD020) through the Rural Development Administration, the Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

scholarly journals Oxymatrine Ameliorates Memory Impairment in Diabetic Rats by Regulating Oxidative Stress and Apoptosis: Involvement of NOX2/NOX4

2020 ◽  
Vol 2020 ◽  
pp. 1-15
Author(s):  
Yongpan Huang ◽  
Xinliang Li ◽  
Xi Zhang ◽  
Jiayu Tang
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Brain Injury ◽  
Western Blotting ◽  
Caspase 3 ◽  
Memory Function ◽  
Diabetic Rats ◽  
Oxygen Species ◽  
Expression Levels

Oxymatrine (OMT) is the major quinolizidine alkaloid extracted from the root of Sophora flavescens Ait and has been shown to exhibit a diverse range of pharmacological properties. The aim of the present study was to investigate the role of OMT in diabetic brain injury in vivo and in vitro. Diabetic rats were induced by intraperitoneal injection of a single dose of 65 mg/kg streptozotocin (STZ) and fed a high-fat and high-cholesterol diet. Memory function was assessed using a Morris water maze test. A SH-SY5Y cell injury model was induced by incubation with glucose (30 mM/l) to simulate damage in vitro. The serum fasting blood glucose, insulin, serum S100B, malondialdehyde (MDA), and superoxide dismutase (SOD) levels were analyzed using commercial kits. Morphological changes were observed using Nissl staining and electron microscopy. Cell apoptosis was assessed using Hoechst staining and TUNEL staining. NADPH oxidase (NOX) and caspase-3 activities were determined. The effects of NOX2 and NOX4 knockdown were assessed using small interfering RNA. The expression levels of NOX1, NOX2, and NOX4 were detected using reverse transcription-quantitative PCR and western blotting, and the levels of caspase-3 were detected using western blotting. The diabetic rats exhibited significantly increased plasma glucose, insulin, reactive oxygen species (ROS), S-100B, and MDA levels and decreased SOD levels. Memory function was determined by assessing the percentage of time spent in the target quadrant, the number of times the platform was crossed, escape latency, and mean path length and was found to be significantly reduced in the diabetic rats. Hyperglycemia resulted in notable brain injury, including histological changes and apoptosis in the cortex and hippocampus. The expression levels of NOX2 and NOX4 were significantly upregulated at the protein and mRNA levels, and NOX1 expression was not altered in the diabetic rats. NOX and caspase-3 activities were increased, and caspase-3 expression was upregulated in the brain tissue of diabetic rats. OMT treatment dose-dependently reversed behavioral, biochemical, and molecular changes in the diabetic rats. In vitro, high glucose resulted in increases in reactive oxygen species (ROS), MDA levels, apoptosis, and the expressions of NOX2, NOX4, and caspase-3. siRNA-mediated knockdown of NOX2 and NOX4 decreased NOX2 and NOX4 expression levels, respectively, and reduced ROS levels and apoptosis. The results of the present study suggest that OMT alleviates diabetes-associated cognitive decline, oxidative stress, and apoptosis via NOX2 and NOX4 inhibition.

Supplementation of kaempferol to in vitro maturation medium regulates oxidative stress and enhances subsequent embryonic development in vitro

Zygote ◽  
2019 ◽  
Vol 28 (1) ◽  
pp. 59-64
Author(s):  
Yuhan Zhao ◽  
Yongnan Xu ◽  
Yinghua Li ◽  
Qingguo Jin ◽  
Jingyu Sun ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Treated Group ◽  
Control Group ◽  
Oxygen Species

SummaryKaempferol (KAE) is one of the most common dietary flavonols possessing biological activities such as anticancer, anti-inflammatory and antioxidant effects. Although previous studies have reported the biological activity of KAE on a variety of cells, it is not clear whether KAE plays a similar role in oocyte and embryo in vitro culture systems. This study investigated the effect of KAE addition to in vitro maturation on the antioxidant capacity of embryos in porcine oocytes after parthenogenetic activation. The effects of kaempferol on oocyte quality in porcine oocytes were studied based on the expression of related genes, reactive oxygen species, glutathione and mitochondrial membrane potential as criteria. The rate of blastocyst formation was significantly higher in oocytes treated with 0.1 µm KAE than in control oocytes. The mRNA level of the apoptosis-related gene Caspase-3 was significantly lower in the blastocysts derived from KAE-treated oocytes than in the control group and the mRNA expression of the embryo development-related genes COX2 and SOX2 was significantly increased in the KAE-treated group compared with that in the control group. Furthermore, the level of intracellular reactive oxygen species was significantly decreased and that of glutathione was significantly increased after KAE treatment. Mitochondrial membrane potential (ΔΨm) was increased and the activity of Caspase-3 was significantly decreased in the KAE-treated group compared with that in the control group. Taken together, these results suggested that KAE is beneficial for the improvement of embryo development by inhibiting oxidative stress in porcine oocytes.

Melatonin delivery by nanocapsules during in vitro bovine oocyte maturation decreased the reactive oxygen species of oocytes and embryos

2016 ◽  
Vol 63 ◽  
pp. 70-81 ◽  
Author(s):  
Mariana Härter Remião ◽  
Caroline Gomes Lucas ◽  
William Borges Domingues ◽  
Tony Silveira ◽  
Nathaniele Nebel Barther ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Oocyte Maturation ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Bovine Oocyte

Tretinoin-loaded lipid-core nanocapsules decrease reactive oxygen species levels and improve bovine embryonic development during in vitro oocyte maturation

2015 ◽  
Vol 58 ◽  
pp. 131-139 ◽  
Author(s):  
Caroline Gomes Lucas ◽  
Mariana Härter Remião ◽  
Eliza Rossi Komninou ◽  
William Borges Domingues ◽  
Cristina Haas ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Embryonic Development ◽  
Oocyte Maturation ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Lipid Core ◽  
In Vitro Oocyte Maturation

scholarly journals Elevation of MPF and MAPK gene expression, GSH content and mitochondrial distribution quality induced by melatonin promotes porcine oocyte maturation and development in vitro

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9913
Author(s):  
Zimo Zhao ◽  
Ling Yang ◽  
Dan Zhang ◽  
Zi Zheng ◽  
Ning Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Developmental Potential ◽  
Expression Levels ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Maturation Rate ◽  
Mitochondrial Distribution ◽  
Mapk Gene

The MPF and MAPK genes play crucial roles during oocyte maturation processes. However, the pattern of MPF and MAPK gene expression induced by melatonin (MT) and its correlation to oocyte maturation quality during the process of porcine oocyte maturation in vitro remains unexplored. To unravel it, in this study, we cultured the porcine oocytes in maturation medium supplemented with 0, 10−6, 10−9, and 10−12 mol/L melatonin. Later, we analyzed the MPF and MAPK gene expression levels by RT-PCR and determined the maturation index (survival and maturation rate of oocytes). The GSH content in the single oocyte, and cytoplasmic mitochondrial maturation distribution after porcine oocyte maturation in vitro was also evaluated. We also assessed the effects of these changes on parthenogenetic embryonic developmental potential. The oocytes cultured with 10−9mol/L melatonin concentration showed higher oocyte maturation rate, and MPF and MAPK genes expression levels along with better mitochondrial distribution than the 0, 10−6, and 10−12 mol/L melatonin concentrations (p < 0.05). No significant difference was observed in the survival rates when the oocytes were cultured with different melatonin concentrations. The expression of the MPF gene in the oocytes cultured with 10−6 mol/L melatonin was higher than with 10−12 and 0 mol/L melatonin, and the expression of the MAPK gene in 10−6 and 10−12 group was higher than the control (p < 0.05). As far as the embryonic developmental potential is concerned, the cleavage and blastocyst rate of oocytes cultured with 10−6 and 10−9 mol/L melatonin was significantly higher than the 10−12 mol/L melatonin and control. In conclusion, 10−9–10−6 mol/L melatonin significantly induced the MPF and MAPK gene expression; besides, it could also be correlated with GSH content of single oocyte, mitochondrial maturation distribution, and the first polar body expulsion. These changes were also found to be associated with parthenogenetic embryo developmental potential in vitro.

172 THE APOPTOTIC EFFECTS OF BISPHENOL A-INDUCED MITOCHONDRIAL-DERIVED REACTIVE OXYGEN SPECIES ON MATURATION OF PORCINE OOCYTES IN VITRO

2017 ◽  
Vol 29 (1) ◽  
pp. 194
Author(s):  
S.-Y. Park ◽  
H.-J. Park ◽  
J.-W. Kim ◽  
J.-Y. Park ◽  
S.-G. Yang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Critical Role ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Meiotic Maturation ◽  
Control Group ◽  
Ros Production ◽  
Maturation In Vitro ◽  
Porcine Oocyte

Bisphenol A (BPA) is well known as oestrogen-like chemical and it is widely used in plastic products. Many studies have reported that BPA exposure has a well-known toxicity effect on reproduction function, such as reducing the number of ovulated oocytes, oocyte quality, and maturation rate. Recently, BPA induced mitochondrial-derived reactive oxygen species (mito-ROS) and disrupted mitochondrial homeostasis by increasing of superoxide anions production. In this study, we investigated how the regulation of mito-ROS production may play a critical role in meiotic maturation and expansion of cumulus cells during the in vitro maturation progression of porcine oocytes. Furthermore, we investigated the toxicity effect of BPA exposure on mitochondrial functions and mito-ROS production during porcine oocyte maturation in vitro. All results were analysed using a 1-way ANOVA followed by Bonferroni’s and Tukey’s Multiple Comparison Test and t-tests. First, porcine oocytes were matured in NCSU-23 medium supplemented with BPA (50, 75, and 100 µM) for 44 h. Our results indicated that the rates of matured oocytes were significantly decreased by BPA exposure in a dose-dependent manner (69.4 ± 5.1, 50.9 ± 6.3, and 29.9 ± 5.8% for BPA treatments of 50, 75, and 100 μM) compared with control group (70.2 ± 7.8%; P < 0.05). Next, we confirmed the secretion functions of oocyte and cumulus cell of cumulus-oocyte complex (COC) and ROS production. Cumulus cell secretion factors (has2, tnfaip6, and cx37) mRNA expression in COC were decreased in the BPA-treated (75 µM) group. In addition, mRNA expressions of mitochondrial-specific antioxidant enzymes (sod2, P < 0.001; prdx3, P < 0.01; prdx5, P < 0.001) and mitochondrial apoptosis genes (bax and caspase-3, P < 0.01) were significantly increased in COC of the BPA-treated (75 µM) group. We measured mitochondrial membrane potential and mito-ROS production using JC-1 analysis and Mito-SOX staining, respectively. The BPA treatment caused a rapid decrease of mitochondrial membrane potential maintenance and increase of mito-ROS production in porcine COC. Moreover, mitochondrial-specific ROS scavenger, Mito-Tempo (0.1 µM) treatment was significantly increased the meiotic maturation of porcine oocytes compared with control group (78.5 ± 3.5 v. 65.8 ± 5.0%; P < 0.05). Based on these results, we first confirmed that BPA exposure reduces the meiotic maturation and cumulus cells expansion of COC by increasing mito-ROS production during porcine oocyte maturation in vitro. Therefore, controlling of mito-ROS for mitochondrial function maintenance and apoptosis plays a critical role in improving porcine oocyte maturation in vitro. This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01117604) and the Bio-industry Technology Development Program (316037–04–1-HD020) through the Rural Development Administration, the Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

scholarly journals PSI-21 The effects of kuromanin supplementation during oocyte maturation on the in vitro production of pig embryos

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 242-243
Author(s):  
Miranda Mentler ◽  
Emma Hicks ◽  
Brian D Whitaker
Keyword(s):  
Reactive Oxygen Species ◽  
Oocyte Maturation ◽  
Reactive Oxygen Species Generation ◽  
Reactive Oxygen ◽  
The Other ◽  
Oxygen Species ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Pronuclear Formation

Abstract The imbalance of reactive oxygen species levels and antioxidants impact oocyte matruation. Elderberries (Sambucus sp.) have been identified as possessing the ability to alleviate oxidative stress. An antioxidant class found in elderberry is anthocyanidin, which includes kuromanin. The objective of this study was to determine the effects of kuromanin supplementation to the media during oocyte maturation. Oocytes (n = 692, r=3) were supplemented with 100 or 200 μM kuromanin during 40-44 h of maturation and then evaluated at the end of maturation for the formation of reactive oxygen species, fertilization characteristics, and rates of embryonic cleavage and blastocyst development at 48 h and 144 h after IVF, respectively. There were no significant differences between no kuromanin supplementation and 100 μM kuromanin supplementation when comparing reactive oxygen species generation at the end of oocyte maturation. Supplementation of 200 μM kuromanin significantly increased (P < 0.05) reactive oxygen species generation in oocytes compared to the other groups. There were no significant differences between no kuromanin supplementation and 100 μM kuromanin supplementation when comparing penetration and polyspermic penetration rates and male pronuclear formation. Supplementation of 200 μM kuromanin significantly decreased (P < 0.05) penetration and polyspermic penetration rates and male pronuclear formation compared to the other groups. There were no significant differences between no kuromanin supplementation and 100 μM kuromanin supplementation when comparing the percentage of cleaved embryos by 48 h after IVF and the percentage of those reaching the blastocyst stage by 144 h after IVF. Supplementation of 200 μM kuromanin significantly decreased (P < 0.05) the cleavage rates by 48 h after IVF and the blastocyst formation rates by 144 h after IVF compared to all other treatment groups. These results indicate that supplementing 200 μM kuromanin is detrimental to oocyte maturation and lower levels (100 μM) do not have a significant effect compared to not supplementing the oocytes when evaluating in vitro fertilization and early embryonic development characteristics.

scholarly journals PSI-22 The effects of cyanidin supplementation during oocyte maturation on the in vitro production of pig embryos

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 243-243
Author(s):  
Emma Hicks ◽  
Miranda Mentler ◽  
Brian D Whitaker
Keyword(s):  
Reactive Oxygen Species ◽  
Oocyte Maturation ◽  
Reactive Oxygen Species Generation ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
In Vitro Production ◽  
Treatment Groups ◽  
The Media ◽  
Pronuclear Formation

Abstract Oxidative stress has a negative effect on embryonic development during the in vitro production of pig embryos. The imbalance of reactive oxygen species and antioxidants impact oocyte maturation. Berries from the elder plant (Sambucus sp.) have been identified as containing high levels of a broad spectrum of antioxidants. One of the predominant antioxidant classes of compounds found in elderberry extract is anthocyanin, which includes the antioxidant cyanidin. Therefore, the objective of this study was to determine the effects of cyanidin supplementation to the media during oocyte maturation. Oocytes (n = 815, r=3) were supplemented with 100 or 200 μM cyanidin during 40–44 h of maturation and then evaluated at the end of maturation for the formation of reactive oxygen species, fertilization characteristics, and rates of embryonic cleavage and blastocyst development at 48 h and 144 h after IVF, respectively. There were no significant differences between no cyanidin supplementation and 200 μM cyanidin supplementation when comparing reactive oxygen species generation at the end of oocyte maturation. Supplementation of 100 μM cyanidin significantly decreased (P < 0.05) reactive oxygen species generation in oocytes compared to the other groups. There were no significant differences between no cyanidin supplementation and 200 μM cyanidin supplementation when comparing penetration and polyspermic penetration rates and male pronuclear formation. Supplementation of 100 μM cyanidin significantly decreased (P < 0.05) polyspermic penetration rates and significantly increased (P < 0.05) male pronuclear formation rates compared to the other groups. There were no significant differences between the treatment groups when comparing the percentage of cleaved embryos by 48 h after IVF. Supplementation of 100 μM cyanidin significantly increased (P < 0.05) the blastocyst formation rates by 144 h after IVF compared to all other treatment groups. These results indicate that supplementing 100 μM cyanidin to the media during oocyte maturation reduces reactive oxygen species formation and improves in vitro fertilization and early embryonic development in pigs.

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