scholarly journals Exposure of Triclosan in Porcine Oocyte Leads to Superoxide Production and Mitochondrial-Mediated Apoptosis during In Vitro Maturation

2020 ◽  
Vol 21 (9) ◽  
pp. 3050 ◽  
Author(s):  
Hyo-Jin Park ◽  
Bong-Seok Song ◽  
Jin-Woo Kim ◽  
Seul-Gi Yang ◽  
Sun-Uk Kim ◽  
...  
Keyword(s):  
Antioxidant Enzymes ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Superoxide Production ◽  
Mrna Levels ◽  
Female Reproduction ◽  
Mitochondrial Superoxide ◽  
Porcine Oocyte

While triclosan (TCS) exerts detrimental effects on female reproduction, the effect of TCS-derived toxins on porcine oocytes during in vitro maturation (IVM) is unclear. This study investigated the effects of TCS on mitochondrion-derived reactive oxygen species (ROS) production and apoptosis pathways during porcine oocyte maturation. Porcine oocytes were treated with TCS (1, 10, and 100 μM) and triphenylphosphonium chloride (Mito-TEMPO; 0.1 μM), and matured cumulus oocyte complexes (COCs) were stained with orcein, dichlorofluorescein diacetate (DCF-DA), and Mito-SOX. Proteins and mRNA levels of factors related to cumulus expansion and mitochondrion-mediated apoptosis and antioxidant enzymes were analyzed by western blotting and reverse-transcription polymerase chain reaction (RT-PCR), respectively. Meiotic maturation and cumulus cell expansion significantly decreased for COCs after TCS treatment along with an increase in mitochondrial superoxide levels at 44 h of IVM. Further, mitochondrion-related antioxidant enzymes and apoptosis markers were significantly elevated in porcine COCs following TCS-mediated oxidative damage. The protective effect of Mito-TEMPO as a specific superoxide scavenger from TCS toxin improved the maturation capacity of porcine COCs. Mito-TEMPO downregulated the mitochondrial apoptosis of TCS-exposed porcine COCs by reducing superoxide level. In conclusion, our data demonstrate that TCS mediates toxicity during porcine oocyte maturation through superoxide production and mitochondrion-mediated apoptosis.

scholarly journals Melatonin Improves Oocyte Maturation and Mitochondrial Functions by Reducing Bisphenol A-Derived Superoxide in Porcine Oocytes In Vitro

2018 ◽  
Vol 19 (11) ◽  
pp. 3422 ◽  
Author(s):  
Hyo-Jin Park ◽  
Soo-Yong Park ◽  
Jin-Woo Kim ◽  
Seul-Gi Yang ◽  
Min-Ji Kim ◽  
...  
Keyword(s):  
Bisphenol A ◽  
Oocyte Maturation ◽  
Cell Damage ◽  
Cumulus Cell ◽  
Superoxide Production ◽  
Positive Effects ◽  
Mitochondrial Functions ◽  
Porcine Oocyte ◽  
Antioxidative Effects

Bisphenol A (BPA) is synthetic organic compound that exhibits estrogen-like properties and it induces mitochondrial superoxide production. Melatonin (Mela) protects against BPA-mediated cell damage and apoptosis. However, the antioxidative effects of Mela against BPA-induced superoxide production in porcine oocytes are still not known. In this study, we investigated the antioxidative effects of Mela against BPA-derived superoxide on oocyte maturation in pigs. To investigate the effects of the superoxide specific scavenger, Mito-TEMPO, on porcine oocyte maturation in response to BPA exposure apoptosis proteins, we treated the oocytes with Mito-TEMPO (0.1 µM) after pre-treating them with BPA (75 µM) for 22 h. As expected, the reduction in meiotic maturation and cumulus cell expansion of cumulus-oocyte-complexes (COCs) in the BPA (75 µM) treated group was recovered (p < 0.01) by treatment with Mito-TEMPO (0.1 µM). An increase in the levels of mitochondrial apoptotic proteins (AIF, cleaved Cas 3 and cleaved Parp1) in response to BPA-induced damage was also reduced by Mito-TEMPO treatment in porcine COCs. Interestingly, we confirmed the positive effects of Mela with respect to superoxide production upon BPA exposure during oocyte maturation and also confirmed the reduction in mitochondrial apoptosis in Mela (0.1 µM)-treated porcine COCs. These results provide evidence for the first time that antioxidative effects of Mela on BPA-derived superoxide improve porcine oocyte maturation.

265 HYALURONAN SYNTHESIS ABILITY OF PORCINE CUMULUS - OOCYTE COMPLEXES DERIVED FROM SMALL FOLLICLES

2013 ◽  
Vol 25 (1) ◽  
pp. 280
Author(s):  
M. Nakakoji ◽  
H. Funahashi
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Cumulus Expansion ◽  
Elisa Kit ◽  
Porcine Oocyte ◽  
Post Hoc ◽  
Metaphase Ii Oocytes

The degree of cumulus expansion, an important step in oocyte maturation, of porcine cumulus–oocyte complexes (COC) derived from small follicles (SF: 1 to 2 mm in diameter) is known to be lower than those derived from middle follicles (MF: 3 to 6 mm in diameter). The objective of this study was to compare the abilities of hyaluronan (HA) synthesis of COC from SF and MF. Furthermore, the effect of oestradiol during pre-incubation of COC on proliferation of the cumulus cells was examined. Cumulus–oocyte complexes from SF and MF of porcine ovaries were cultured for in vitro maturation [IVM, in modified porcine oocyte medium (Yoshioka et al. 2008 J. Reprod. Dev. 54, 208–213) supplemented with 50 µM β-mercaptoethanol, 10 IU mL–1 of eCG, 10 IU mL–1 of hCG, and 1 mM dbcAMP for 20 h and then in the fresh medium without those supplements for another 24 h]. Hyaluronan production was quantified at 20 h after the start of IVM with a commercial HA-ELISA kit (20 COC/tube × 4 times). The number of cumulus cells was assessed 0 and 20 h after the start of IVM (50 COC × 4 times). Furthermore, proliferation of cumulus cells was examined after pre-culture of COC (n = 40 COC × 5 times) in modified porcine oocyte medium with various concentrations of oestradiol (0, 0.1, 1, and 10 ng mL–1) for 6 h. Statistical analyses of results from 4 to 5 replicated trials were performed by ANOVA with a Bonferroni-Dunn post-hoc test (significance, P < 0.05). The degree of cumulus expansion of COC from MF (n = 152) was higher than that of COC from SF (n = 156). The incidence of metaphase-II oocytes was significantly lower in COC from SF (n = 133; 48.9%) than in COC from MF (n = 148; 74.7%). The HA content of COC was higher in those from MF (20.8 µg/COC) than in those from SF (10.8 µg/COC), whereas the content per cumulus cell was not different because the numbers of cumulus cells at 0 and 20 h were also higher in COC (n = 200 in each group) from MF (3.0 × 103 and 3.3 × 103 cells, respectively) than from SF (2.0 × 103 and 2.5 × 103 cells, respectively). Cumulus cells proliferated significantly in the presence of oestradiol, regardless of the concentration, during pre-incubation for 6 h (2.5 to 2.8 × 103 cells), as compared with the oestradiol-free controls (2.2 × 103 cells). These results demonstrate that the different abilities of cumulus expansion between COC (n = 200 in each group) from SF and MF may be due to the number of cumulus cells per COC. Pre-incubation in the presence of oestradiol stimulates the proliferation of cumulus cells and may improve the oocyte maturation of COC derived from SF.

254 9-cis RETINOIC ACID INHIBITS CUMULUS CELL APOPTOSIS DURING IN VITRO MATURATION OF BOVINE OOCYTES THROUGH INHIBITION OF AP-1 PATHWAY

2011 ◽  
Vol 23 (1) ◽  
pp. 225
Author(s):  
G. K. Deb ◽  
S. R. Dey ◽  
J. I. Bang ◽  
S. J. Cho ◽  
T. H. Kwon ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Tumor Necrosis Factor ◽  
Oocyte Maturation ◽  
Tumor Necrosis ◽  
In Vitro Maturation ◽  
Tumor Necrosis Factor Α ◽  
Cumulus Cell ◽  
Factor Α ◽  
Necrosis Factor

Cumulus cells (CC) play a critical role in oocyte maturation and fertilization via gap junctions. The oocyte itself maintains CC health to favour oocyte maturation via the secretion of paracrine growth factors. However, the antiapoptotic effects of oocyte-secreted factors follow a gradient from the site of the oocytes. Moreover, degrees of CC apoptosis are inversely related to the in vitro embryo development. Therefore, inhibition of CC apoptosis is important for efficient in vitro embryo development. The beneficial effects of retinoic acid (RA) during in vitro embryo production are well known in different species. However, the effect of RA on CC apoptosis is yet to be elucidated. All-trans RA and 9-cis RA are the natural components of retinoids, and all-trans RA are metabolized to 9-cis RA for physiological function. Therefore, the objective of the present study was to evaluate the effect of 9-cis RA on the mechanism for inhibition of apoptosis in CC. Slaughterhouse cumulus–oocyte complexes (COC) were matured in vitro in TCM-199-based in vitro maturation medium containing 0 or 5 mM 9-cis RA for 23 to 24 h (15 COC/100 μL droplet) at 38.5°C and 5% CO2 in air with maximum humidity. Following in vitro maturation, COC of a droplet were fixed in 4% paraformaldehyde for TUNEL staining using In Situ Cell Death Detection Kit (Roche, Budapest, Hungary). The proportion of apoptotic cells was estimated using Olympus Soft Imaging Solutions GmBH (Olympus, Münster, Germany). The COC of the remaining droplet were denuded. The CC were frozen and stored at –80°C. The CC of 3 different cultures were pooled, and total RNA was extracted using RNeasy Mini Kit (Qiagen, Valencia, CA, USA). Total RNA was reverse transcribed into cDNA using Omniscript Reverse Transcription kit (Qiagen). Relative expression of candidate genes was quantified using SYBER green real-time PCR with ΔΔ CT method. The expression was normalized against β-actin, glyceraldehyde 3-phosphate dehydrogenase, and 18s rRNA genes expression. The PCR efficiencies were calculated using relative calibration curves following 10-fold dilution series at 5 measuring points. Data were analysed for one-way ANOVA. The proportion of apoptotic cells was low in the 9-cis RA group (1.3 v. 3.3% of total CC; P < 0.05). Expression of tumor necrosis factor-α (11.1 v. 1.0; P < 0.001), caspase9 (2.0 v. 1.0; P < 0.01), and caspase3 (2.1 v. 1.0; P < 0.001) genes was down-regulated in the 9-cis RA group, whereas expression of Bcl2 gene was increased (1.0 v. 2.6 fold; P < 0.05). Moreover, the expression of c-fos gene of AP-1 pathway was down-regulated (1.9 v. 1 fold; P < 0.05) in the 9-cis RA group. Retinoic acid suppressed the expression of NF-kB, which in turn inhibits tumor necrosis factor-α-mediated caspase activity. However, the expression of NF-kB in CC was not affected by 9-cis RA (1.1 v. 1.0; P > 0.05). In conclusion, the present study indicated that 9-cis RA may inhibit cumulus cell apoptosis through suppression of AP-1 pathway. This work was partly supported by a scholarship from the BK21 program, the KRF (KRF-2008-211-F00011), the IPET (108068-03-1-SB010), and the KOSEF (10525010001-05N2501-00110).

188 IMPROVEMENT OF IN VITRO CANINE OOCYTE MATURATION BY OVIDUCTAL SECRETOME

2017 ◽  
Vol 29 (1) ◽  
pp. 202 ◽  
Author(s):  
A. Lange-Consiglio ◽  
C. Perrini ◽  
P. Esposti ◽  
F. Cremonesi
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Follicular Development ◽  
Cumulus Cells ◽  
Chi Square ◽  
Cell Layers ◽  
Hoechst Staining

The in vitro maturation of canine oocyte is problematic because it is difficult to reproduce the oviducal microenvironment where the in vivo maturation occurs. Because cells are able to communicate with each other by paracrine action, oviducal cells could be in vitro cultivated to obtain the conditioned medium (CM) consisting of soluble factors and microvesicles (MV), which represent a carrier for nonsoluble molecules including microRNA. The aim of the present work was to investigate the effect of the addition of CM or MV, secreted by oviducal cells, to the canine in vitro maturation medium. To generate CM, cells from oviducts of 3 animals in late oestrus were cultured for 5 days at 38.5°C in a humidified atmosphere of 5% CO2. Supernatants were collected, pooled, centrifuged at 2500 × g, and stored at −80°C. Microvesicles were obtained by ultracentrifugation of CM at 100,000 × g for 1 h at 4°C and measured for concentration and size by a Nanosight instrument. Ovaries were obtained from 50 healthy domestic bitches (1–4 years old) of different breeds that underwent ovariectomy regardless of the oestrous cycle. Cumulus-oocyte complexes were released by slicing the ovarian cortex with a scalpel blade, and only Grade 1 cumulus-oocyte complexes (darkly granulated cytoplasm and surrounded by 3 or more compact cumulus cell layers) 110 to 120 µm in diameter were selected for culture. Maturation was performed at 38.5°C in a humidified atmosphere of 5% CO2 and 5% of O2 in bi-phasic systems: 24 h in SOF with 5.0 μg mL−1 of LH followed by 48 h in SOF supplemented with 10% of oestrous bitch serum and 10% CM or 50, 75, 100, or 150 × 106 MV mL−1 labelled with PKH-26. Control was the same medium without CM or MV. Oocytes were observed under a fluorescent microscope to detect metaphase II (MII), by Hoechst staining, and the incorporation of MV. Statistical analysis was performed by chi-square test. Results show that canine oviducal cells secreted MV of 234 ± 23 nm in size, underling that these MV fall within the shedding vesicles category. The incorporation of labelled MV occurred at first in cumulus cells, at 48 h of maturation, and then, at 72 h, in oocyte cytoplasm. These MV had a positive effect on maturation rate (MII) at the concentration of 75 and 100 × 106 MV mL−1 compared with CM and control (20.34 and 21.82 v. 9.09 and 3.95%, respectively). The concentration of 150 × 106 MV mL−1 provided only 9.26% of MII. To understand the role of MV, we assessed the expression of 3 microRNA (miRNA-30b, miR-375, and miR-503) that are involved in some key pathways (WNT, MAPK, ERbB, and TGFβ) regulating follicular development and meiotic resumption. The lower rate of MII with the higher concentration of MV is possibly due to the high level of miR-375, which recent literature shows to suppress the TGFβ pathway, leading to impaired oocyte maturation. In conclusion, the oviducal MV, or specific microRNA, are involved in cellular trafficking during oocyte maturation, and their possible use in vitro could facilitate the exploitation of canine reproductive biotechnologies.

scholarly journals Butylparaben Is Toxic to Porcine Oocyte Maturation and Subsequent Embryonic Development Following In Vitro Fertilization

2020 ◽  
Vol 21 (10) ◽  
pp. 3692 ◽  
Author(s):  
Pil-Soo Jeong ◽  
Sanghoon Lee ◽  
Soo-Hyun Park ◽  
Min Ju Kim ◽  
Hyo-Gu Kang ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Oocyte Maturation ◽  
Mitochondrial Function ◽  
Cumulus Cell ◽  
Annexin V ◽  
Ros Generation ◽  
Vitro Fertilization ◽  
Porcine Oocyte ◽  
Mitochondrial Distribution

Parabens are widely used in personal care products due to their antimicrobial effects. Although the toxicity of parabens has been reported, little information is available on the toxicity of butylparaben (BP) on oocyte maturation. Therefore, we investigated the effects of various concentrations of BP (0 μM, 100 μM, 200 μM, 300 μM, 400 μM, and 500 μM) on the in vitro maturation of porcine oocytes. BP supplementation at a concentration greater than 300 μM significantly reduced the proportion of complete cumulus cell expansion and metaphase II oocytes compared to the control. The 300 μM BP significantly decreased fertilization, cleavage, and blastocyst formation rates with lower total cell numbers and a higher rate of apoptosis in blastocysts compared to the control. The BP-treated oocytes showed significantly higher reactive oxygen species (ROS) levels, and lower glutathione (GSH) levels than the control. BP significantly increased the aberrant mitochondrial distribution and decreased mitochondrial function compared to the control. BP-treated oocytes exhibited significantly higher percentage of γ-H2AX, annexin V-positive oocytes and expression of LC3 than the control. In conclusion, we demonstrated that BP impaired oocyte maturation and subsequent embryonic development, by inducing ROS generation and reducing GSH levels. Furthermore, BP disrupted mitochondrial function and triggered DNA damage, early apoptosis, and autophagy in oocytes.

272 A NEW APPROACH TO IN VITRO MATURATION (IVM) AND EMBRYO IN VITRO PRODUCTION: INDUCED IVM SUBSTANTIALLY IMPROVES EMBRYO YIELD AND PREGNANCY OUTCOMES

2010 ◽  
Vol 22 (1) ◽  
pp. 293
Author(s):  
R. B. Gilchrist ◽  
F. K. Albuz ◽  
J. G. Thompson
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
In Vitro Production ◽  
New Approach ◽  
Fetal Survival ◽  
Wide Range ◽  
Vitro Production

Oocyte in vitro maturation (IVM) is the rate-limiting step in the in vitro production (IVP) of embryos. Oocyte maturation in vivo is a highly orchestrated, induced process, whereby cAMP-mediated meiotic arrest is overridden by the gonadotrophin surge prior to ovulation. However, aspirated oocytes resume maturation spontaneously compromising developmental competence. Hence, we hypothesized that establishing an induced system in vitro would synchronize oocyte-somatic cell communication leading to improved oocyte quality. Abattoir-collected bovine or 129/Sv mouse oocytes were treated for the first 1 to 2 h in vitro (pre-IVM) with the adenylate cyclase activator forskolin (100 μM, 50 μM, respectively) and a nonspecific phosphodiesterase (PDE) inhibitor, IBMX (500 μM, 50 μM), which substantially increased cumulus-oocyte complex (COC) cAMP (bovine, 180 v. 2 fmol/COC, treated v. control; P < 0.001). To maintain oocyte cAMP levels and prevent precocious oocyte maturation, IVM media (VitroMat + BSA) contained an oocyte-specific (type 3) PDE inhibitor, cilostamide (20 μM, 0.1 μM), plus FSH to induce maturation. The net effect of this system (induced-IVM) was to increase oocyte-cumulus cell gap-junctional communication (bovine: 1000 ± 148 v. 340 ± 73 unit, treated v. control; P < 0.05) and to slow meiotic progression through prophase I to metaphase II, extending the normal IVM interval (bovine: 30 v. 24 h, mouse: 22 v. 18 h; treated v. control). FSH was required to complete maturation and FSH-induced maturation was prevented by an epidermal growth factor receptor inhibitor, AG1478 (2.5 μM), demonstrating induced oocyte maturation functions via secondary autocrine signaling within the cumulus cell compartment. These effects on COC functions had profound consequences for oocyte developmental potential. In completely serum-free bovine IVP, induced-IVM more than doubled blastocyst yield (69 v. 27%, treated v. control; P < 0.05) and improved blastocyst quality (186 v. 132 blastomeres). To achieve these rates, the pre-IVM phase, the modified IVM conditions, and delayed IVF were all required. Adapting the system to the mouse, induced-IVM increased blastocyst rate (86 v. 55%, treated v. control; P < 0.05), implantation rate (51 v. 25%; P < 0.01), fetal survival rate (29 v. 5%; P < 0.01) and fetal weight (0.9 v. 0.5 g; P < 0.01). All these embryonic and fetal outcomes in mice were equivalent (P > 0.05) using induced-IVM to levels obtained from in vivo-matured control oocytes (conventional IVF). Data were analyzed by ANOVA. In conclusion, induced-IVM mimics some of the characteristics of oocyte maturation in vivo and substantially improves oocyte developmental outcomes in 2 disparate mammalian species. Adaption of this new approach to clinical/field conditions should lead to new opportunities for a wide range of reproductive biotechnologies. Such a notable increase in IVM efficiency could see IVP as the preferred embryo production technology in future livestock artificial breeding programs. Funded by an Australian Research Council Linkage Grant and Cook Australia. Thanks to M. Sasseville, M. Lane, and D. T. Armstrong.

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 Nucleus, Cytoskeleton, and Mitogen-Activated Protein Kinase p38 Dynamics during In Vitro Maturation of Porcine Oocytes

Animals ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 163
Author(s):  
Payungsuk Intawicha ◽  
Li-Kuang Tsai ◽  
Shih-Ying Yen ◽  
Neng-Wen Lo ◽  
Jyh-Cherng Ju
Keyword(s):  
Oocyte Maturation ◽  
Mammalian Cells ◽  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Mitogen Activated Protein Kinase ◽  
Meiotic Maturation ◽  
Germinal Vesicle Breakdown ◽  
Mitogen Activated Protein ◽  
Porcine Oocyte

The mitogen-activated kinase (MAPK) p38, a member of the MAPK subfamily, is conserved in all mammalian cells and plays important roles in response to various physiologic cues, including mitogens and heat shock. In the present study, MAPK p38 protein expression in porcine oocytes was analyzed during in vitro maturation (IVM) by Western blotting and immunocytochemistry. The levels of p-p38 or activated p38 and p38 expression were at the lowest in the germinal vesicle (GV) stage oocyte, gradually rising at the germinal vesicle breakdown (GVBD) and then reaching a plateau throughout the IVM culture (p < 0.05). Similarly, the expression level of total p38 was also lower in the GV oocyte than in the oocyte of other meiotic stages and uprising after GVBD and remained high until the metaphase III (MII) stage (p < 0.05). In the GV stage, phosphorylated p38 (p-p38) was initially detectable in the ooplasm and subsequently became clear around the nucleus and localized in the ooplasm at GVBD (18 h post-culture). During the metaphase I (MI) and metaphase II (MII) stages, p-p38 was evenly distributed throughout the ooplasm after IVM for 30 or 42 h. We found that the subcellular localization increased in p-p38 expression throughout oocyte maturation (p < 0.05) and that dynamic reorganization of the cytoskeleton, including microfilaments and microtubules, was progressively changed during the course of meiotic maturation which was likely to be associated with the activation or networking of p38 with other proteins in supporting oocyte development. In conclusion, the alteration of p38 activation is essential for the regulation of porcine oocyte maturation, accompanied by the progressive reorganization and redistribution of the cytoskeleton and MAPK p38, respectively, in the ooplasm.

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.

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