scholarly journals The Effects of Polyadenylation Status on MPFs During In Vitro Porcine Oocyte Maturation

2016 ◽  
Vol 39 (5) ◽  
pp. 1735-1745 ◽  
Author(s):  
Huiyu Liu ◽  
Yan Gao ◽  
Bo Zhai ◽  
Hao Jiang ◽  
Yu Ding ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Cyclin B1 ◽  
Spindle Formation ◽  
Large Follicle ◽  
Maturation In Vitro ◽  
First Polar Body ◽  
Chromosome Alignment ◽  
Porcine Oocyte ◽  
Maternal Mrnas

Aims: This study aims to clarify the effects of polyadenylation status on M-phase promoting factors (MPFs) during in vitro porcine oocyte maturation. Methods: In this study, porcine follicular oocytes from large follicles (> 5 millimeter (mm)) and small follicles (< 3 mm) were examined at different follicular developmental stages. The polyadenylation of maternal mRNAs was inhibited by the addition of 3'-deoxyadenosine (3'-da) during the germinal vesicle (GV)(0 h), GV breakdown (GVBD)(18 h), metaphase I (MI)(28 h), and metaphase II (MII) (44 h) stages. In addition, the expression levels and poly-(A) tail lengths of the maternal mRNAs Cyclin B1 and cell division cycle 2 (Cdc2) were determined by real-time quantitative PCR. Immunofluorescence was used to assess spindle formation and chromosome alignment in the examined oocytes. Results: In large-follicle oocytes, the effects of inhibiting polyadenylation caused the percentage of mature to be significantly lower for the treated group than for the untreated group (p < 0.01). 3'-da can significantly improve the rate of small oocyte maturation in vitro and inhibits Cdc2 polyadenylation. Cyclin B1 plays a significant role in promoting the maturation of large-follicle oocytes. Polyadenylation contributes to the formation of dominant follicles and facilitates the selection of dominant follicles. However, the inhibition of adenylation affected spindle formation-related propulsion and chromosome alignment in both large- and small-follicle oocytes. The first polar body could not be extruded in certain large follicles. Conclusions: 3'-da can significantly improve the rate of small oocyte maturation in vitro, but it can also affect spindle formation-related propulsion and chromosome alignment.

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.

Sonic hedgehog promotes porcine oocyte maturation and early embryo development

2009 ◽  
Vol 21 (6) ◽  
pp. 805 ◽  
Author(s):  
Ngoc Tan Nguyen ◽  
David Pei-Cheng Lin ◽  
Shih-Ying Yen ◽  
Jung-Kai Tseng ◽  
Jui-Fen Chuang ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sonic Hedgehog ◽  
Oocyte Maturation ◽  
Calcium Release ◽  
Cyclin B1 ◽  
Early Embryo ◽  
Control Group ◽  
Early Embryo Development ◽  
Porcine Oocyte

In the present study, we investigated the effects of the Sonic hedgehog (Shh) protein on porcine oocyte maturation and early embryo development. Immunohistochemistry showed activation of Shh signalling in cumulus–oocyte complexes (COCs), as reflected by Patched (Ptc), Smoothened (Smo) and Gli1 expression in oocytes, cumulus cells and granulosa cells, particularly those of small follicles (<2 mm in diameter). Western blot analysis showed Smo expression in COCs and in denuded oocytes derived from small and medium (3–7 mm)-sized follicles. Small follicles contained the highest concentration of Shh in follicular fluid compared with medium-sized and large (>7 mm in diameter) follicles. Supplementation with Shh (0.5 or 1 μg mL–1) enhanced oocyte maturation compared with the control group (92.4% and 90.4% v. 81.9%, respectively; P < 0.05). This effect was reversed by the simultaneous addition of cyclopamine (1–2 μm), an Shh inhibitor. Similar to intact COCs, denuded COCs showed enhanced maturation following Shh supplementation. Furthermore, cyclin B1 content, extracellular signal-regulated kinase 1/2 phosphorylation, intracellular calcium release, blastocyst rate and total cell numbers were greater (P < 0.05) in oocytes matured in the presence of 0.5 and 1 μg mL–1 Shh compared with control oocytes. The findings of the present study provide the first evidence that the Shh signalling pathway is active, or at least partially activated, in the porcine ovary and is likely to promote oocyte cytoplasmic and nuclear maturation, as well as subsequent in vitro development, although the underlying mechanisms remain to be elucidated.

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.

Toxic effects and possible mechanisms of hydrogen sulfide and/or ammonia on porcine oocyte maturation in vitro

2018 ◽  
Vol 285 ◽  
pp. 20-26 ◽  
Author(s):  
Lei-Lei Yang ◽  
Yong Zhao ◽  
Shi-Ming Luo ◽  
Jun-Yu Ma ◽  
Zhao-Jia Ge ◽  
...  
Keyword(s):  
Hydrogen Sulfide ◽  
Oocyte Maturation ◽  
Toxic Effects ◽  
Maturation In Vitro ◽  
Porcine Oocyte

GDF8 activates p38 MAPK signaling during porcine oocyte maturation in vitro

2017 ◽  
Vol 101 ◽  
pp. 123-134 ◽  
Author(s):  
Junchul David Yoon ◽  
Seon-Ung Hwang ◽  
Eunhye Kim ◽  
Minghui Jin ◽  
Soochong Kim ◽  
...  
Keyword(s):  
P38 Mapk ◽  
Oocyte Maturation ◽  
Mapk Signaling ◽  
Maturation In Vitro ◽  
Porcine Oocyte

Porcine oocyte maturation in vitro under different conditions

2006 ◽  
Vol 3 (2) ◽  
pp. 95-100
Author(s):  
Hu Jun-He ◽  
Yang Chun-Rong ◽  
Dou Zhong-Ying
Keyword(s):  
Oocyte Maturation ◽  
Culture Medium ◽  
Follicle Stimulating Hormone ◽  
Human Chorionic Gonadotrophin ◽  
Serum Free Medium ◽  
Free Medium ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Pregnant Mare Serum Gonadotrophin

AbstractThe effects of hormone additions at various stages and different basic media, with or without serum, on porcine oocyte maturation in vitro were studied. The results showed that the rate of maturation was not significantly different with three different stages of hormone supplement; the rate of maturation on modified TCM199 medium (54.01%) was higher than that on TCM199 (46.16%) and (47.14%), but these differences were not significant; and the rate of maturation on serum-free medium (67.10%) was significantly higher than that on medium plus serum (52.22%). Therefore, modifed tissue culture medium 199 (mTCM199)+10I U/ml pregnant mare serum gonadotrophin (PMSG)+10I U/ml human chorionic gonadotrophin (hCG)+2.5 IU/ml follicle stimulating hormone (FSH) was a suitable medium for culture of porcine oocytes in vitro, and the rate of maturation was 67.10%.

scholarly journals Progesterone influences cytoplasmic maturation in porcine oocytes developingin vitro

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2454 ◽  
Author(s):  
Bao Yuan ◽  
Shuang Liang ◽  
Yong-Xun Jin ◽  
Jeong-Woo Kwon ◽  
Jia-Bao Zhang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Specific Inhibitor ◽  
Blastocyst Stage ◽  
Female Reproduction ◽  
Nuclear Maturation ◽  
First Polar Body ◽  
Porcine Oocyte ◽  
Cytoplasmic Maturation

Progesterone (P4), an ovarian steroid hormone, is an important regulator of female reproduction. In this study, we explored the influence of progesterone on porcine oocyte nuclear maturation and cytoplasmic maturation and developmentin vitro. We found that the presence of P4 during oocyte maturation did not inhibit polar body extrusions but significantly increased glutathione and decreased reactive oxygen species (ROS) levels relative to that in control groups. The incidence of parthenogenetically activated oocytes that could develop to the blastocyst stage was higher (p< 0.05) when oocytes were exposed to P4 as compared to that in the controls. Cell numbers were increased in the P4-treated groups. Further, the P4-specific inhibitor mifepristone (RU486) prevented porcine oocyte maturation, as represented by the reduced incidence (p< 0.05) of oocyte first polar body extrusions. RU486 affected maturation promoting factor (MPF) activity and maternal mRNA polyadenylation status. In general, these data show that P4 influences the cytoplasmic maturation of porcine oocytes, at least partially, by decreasing their polyadenylation, thereby altering maternal gene expression.

scholarly journals 313IN VITRO MATURATION AND FERTILIZATION OF OOCYTES COLLECTED FROM UNSTIMULATED MACACA NEMISTRINA OVARIES

2004 ◽  
Vol 16 (2) ◽  
pp. 276
Author(s):  
E.S. Hayes ◽  
E.C. Curnow
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Pantothenic Acid ◽  
Macaca Nemestrina ◽  
Chi Square ◽  
Maturation In Vitro ◽  
First Polar Body ◽  
Chi Square Analysis ◽  
Polar Body Extrusion

Reports describing the IVM of Macaca nemestrina (Mn) oocytes are limited (Cranfield MR et al. 1989 Zoo. Biol. (Supp. 1), 33). The use of gonadotrophins (Gnt) for IVM of non-human primate (NHP) oocytes is common but the concentrations used are often high (8–40IUmL−1) and the species of origin and biological activity of Gnt varies (Schramm RD and Paprocki AM, 2000 Hum. Reprod. 15, 2411). We have compared two different IVM systems with human Gnt on maturation and fertilization of oocytes collected from unstimulated Mn ovaries (n=6–10 animals). Oocytes were subjected to IVM in modified (minus PVA and pantothenic acid, plus 20 amino acids) HECM−10+15% FCS (Zheng P et al., 2001 Mol. Reprod. Dev. 58, 348) for a) 36h in the presence (mHECM+36, n=322) or absence (mHECM−36, n=99) of FSH and LH applied sequentially (FSH 1IUmL−1 0–24h; 10IUmL−1 FSH and LH 24–36h) or b) 24h in the presence (mHECM+24, n=119) or absence (mHECM−24, n=56) of static concentrations of Gnt (FSH and LH 1IUmL−1 0–24h; no Gnt 24–30h). Oocytes exhibiting first polar body extrusion at 36 and 30h were recorded as mature (MII) and subjected to IVF in HTF+BSA (3mg mL−1) with Mn sperm pretreated with 1.0mM caffeine and 0.1mM dbcAMP. Fertilized oocytes (pronuclei and/or 2nd polar body extrusion) were cultured in sequential culture medium for 48h, assessed for cleavage and either fixed or frozen. Proportional data (mature/total, fertilized/mature or cleaved/fertilized) were compared by chi-square analysis and are reported as percentages. Oocytes cultured in mHECM+36 and mHECM−36 exhibited similar rates of GVBD (58.7% v. 53.5%) but the percentage of MII oocytes was significantly higher (P=0.0244) in mHECM+36 (41.3%) v. mHECM−36 (28.3%). Fertilization rates were comparable between mHECM+36 (61.5%) and mHECM−36 (60.9%), whereas cleavage rates were significantly higher (P=0.0004) in mHECM+36 (74.6%) v. HECM−36 (21.4%). Oocytes cultured in mHECM+24 and mHECM−24 exhibited similar rates of GVBD (76.5% v. 62.5%) but the proportion of MII oocytes was significantly higher (P=0.0159) in mHECM+24 (55.5%) v. mHECM−24 (35.7%). Fertilization and cleavage rates were comparable between mHECM+24 (58.8% v. 63.3%) and mHECM−24 (50.0% v. 42.8%). A comparison between mHECM+36 and mHECM+24 indicated a significantly lower (P=0.0005) percentage of GV oocytes and a significantly higher (P=0.0096) percentage of MII oocytes in mHECM+24 (23.5% v. 55.5%) compared to mHECM+36 (41.3% v. 41.3%). Fertilization and cleavage rates were not significantly different between mHECM+36 and mHECM+24. Oocyte maturation and fertilization and embryo cleavage were not different for mHECM−36 and mHECM−24 (P=0.3138–0.8202). Mn oocytes exhibit high rates of Gnt-independent GVBD (52.5%–53.5%) and maturation (28.3%–35.7%) in vitro, and maturation rates were improved in Gnt supplemented maturation medium. However, reduced exposure to lower concentrations of FSH and increased exposure to lower concentrations of LH was associated with higher rates of oocyte maturation in vitro. The use of lower concentrations of FSH and LH for reduced periods may improve IVM of NHP oocytes. This work was supported by the Tissue Distribution Program of the WaNPRC (NIH grant # R00166).

scholarly journals Reduction of Mitochondrial Derived Superoxide by Mito-TEMPO Improves Porcine Oocyte Maturation In Vitro

2019 ◽  
Vol 34 (1) ◽  
pp. 10-19 ◽  
Author(s):  
Seul-Gi Yang ◽  
Hyo-Jin Park ◽  
Sang-Min Lee ◽  
Jin-Woo Kim ◽  
Min-Ji Kim ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Maturation In Vitro ◽  
Porcine Oocyte
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