scholarly journals Tributyltin Oxide Exposure During in vitro Maturation Disrupts Oocyte Maturation and Subsequent Embryonic Developmental Competence in Pigs

2021 ◽  
Vol 9 ◽  
Author(s):  
Yue Xiao ◽  
Bao Yuan ◽  
Weiyi Hu ◽  
Jiajia Qi ◽  
Hao Jiang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Iron Homeostasis ◽  
Polar Body ◽  
Developmental Competence ◽  
Limited Information ◽  
Cellular Iron ◽  
Ros Accumulation ◽  
Porcine Oocyte ◽  
Cellular Iron Homeostasis

Tributyltin oxide (TBTO), an organotin compound, has been demonstrated to have toxic effects on several cell types. Previous research has shown that TBTO impairs mouse denuded oocyte maturation. However, limited information is available on the effects of TBTO exposure on livestock reproductive systems, especially on porcine oocytes in the presence of dense cumulus cells. In the present research, we evaluated the effects of TBTO exposure on porcine oocyte maturation and the possible underlying mechanisms. Porcine cumulus-oocyte complexes were cultured in maturation medium with or without TBTO for 42 h. We found that TBTO exposure during oocyte maturation prevented polar body extrusion, inhibited cumulus expansion and impaired subsequent blastocyst formation after parthenogenetic activation. Further analysis revealed that TBTO exposure not only induced intracellular reactive oxygen species (ROS) accumulation but also caused a loss of mitochondrial membrane potential and reduced intracellular ATP generation. In addition, TBTO exposure impaired porcine oocyte quality by disrupting cellular iron homeostasis. Taken together, these results demonstrate that TBTO exposure impairs the porcine oocyte maturation process by inducing intracellular ROS accumulation, causing mitochondrial dysfunction, and disrupting cellular iron homeostasis, thus decreasing the quality and impairing the subsequent embryonic developmental competence of porcine oocytes.

Treatment of allicin improves maturation of immature oocytes and subsequent developmental ability of preimplantation embryos

Zygote ◽  
2017 ◽  
Vol 25 (4) ◽  
pp. 480-488 ◽  
Author(s):  
Sang-Gi Jeong ◽  
Seung-Eun Lee ◽  
Yun-Gwi Park ◽  
Yeo-Jin Son ◽  
Min-Young Shin ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Treated Group ◽  
Developmental Competence ◽  
Mitogen Activated Protein Kinase ◽  
Preimplantation Embryos ◽  
Maturation Medium ◽  
Mitogen Activated Protein ◽  
Porcine Oocyte

SummaryAllicin (AL) regulates the cellular redox, proliferation, viability, and cell cycle of different cells against extracellular-derived stress. This study investigated the effects of allicin treatment on porcine oocyte maturation and developmental competence. Porcine oocytes were cultured in medium supplemented with 0 (control), 0.01, 0.1, 1, 10 or 100 μM AL, respectively, during in vitro maturation (IVM). The rate of polar body emission was higher in the 0.1 AL-treated group (74.5% ± 2.3%) than in the control (68.0% ± 2.6%) (P < 0.1). After parthenogenetic activation, the rates of cleavage and blastocyst formation were significantly higher in the 0.1 AL-treated group than in the control (P < 0.05). The reactive oxygen species level at metaphase II did not significantly differ among all groups. In matured oocytes, the expression of both BAK and CASP3, and BIRC5 was significantly lower and higher, respectively, in the 0.1 AL-treated group than in the control. Similarly, the expression of BMP15 and CCNB1, and the activity of phospho-p44/42 mitogen-activated protein kinase (MAPK), significantly increased. These results indicate that supplementation of oocyte maturation medium with allicin during IVM improves the maturation of oocytes and the subsequent developmental competence of porcine oocytes.

167 DEVELOPMENTAL COMPETENCE OF PORCINE OOCYTES DERIVED FROM SMALL- OR MEDIUM-SIZED FOLLICLES AND DENUDED OF CUMULUS CELLS BEFORE AND DURING IN VITRO MATURATION

2017 ◽  
Vol 29 (1) ◽  
pp. 192
Author(s):  
P. Ferré ◽  
K. X. Nguyen ◽  
T. Wakai ◽  
H. Funahashi
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Blastocyst Formation ◽  
First Polar Body

This experiment was undertaken to assess the meiotic and developmental competences of oocytes derived from different sized follicles and denuded of cumulus cells 0, 20, and 44 h after the start of culture for in vitro maturation (IVM). Groups of 60 oocyte-cumulus complexes from small- (SF; <3 mm) and medium-sized follicles (MF; 3–6 mm) were cultured for IVM in porcine oocyte medium with 50 μM β-mercaptoethanol supplemented with 1 mM dibutyryl-cyclic adenosine monophosphate, 10 IU mL−1 of eCG, and 10 IU mL−1 of hCG for 20 h at 39°C and 5% CO2 in air. Then, after washing, they continued culture in fresh β-mercaptoethanol without dibutyryl-cyclic adenosine monophosphate and gonadotropins under the same conditions for another 24 h. At 0, 20, and 44 h of IVM, cumulus cells were removed with 0.1% (wt/vol) hyaluronidase and the denuded oocytes continued IVM culture following the protocol. Mature oocytes with the first polar body were selected, parthenogenetically activated with a single electrical pulse (DC: 1.2 kV/cm, 30 µs), incubated with 4% (wt/vol) BSA and 5 μM cytochalasin B for 4 h, and cultured in porcine zygote medium for 5 days. Cleavage and blastocyst formation rates were observed on Day 2 and 5, respectively. Blastocysts were stained with 4’,6-diamidino-2-phenylindole for cell count assessment. The experiment was replicated 5 times and analysed with a 1- or 2-way ANOVA. If P < 0.05 in ANOVA, a Tukey multiple comparisons test was performed. Regardless of the time of cumulus cell removal, oocytes from MF had significantly higher in rates of maturation, cleavage, and blastocyst rates, as compared with those from SF, whereas there were no significant differences in the cell number of blastocysts between SF and MF (32 v. 34 cells, respectively). When oocytes were denuded before IVM culture, rates of oocyte maturation (37.6% in SF and 50.8% in MF), and blastocyst formation (2.7% in SF and 27.3% in MF) were significantly lower than controls (51.2% in SF and 76% in MF; 25.8% in SF and 48.5% in MF, respectively). When oocytes were denuded 20 h after the start of IVM, oocyte maturation rates were significantly increased (64.1% in SF and 82.5% in MF) as compared with controls, whereas no significant differences were observed in cleavage and blastocyst formation rates in comparison with controls. These results conclude that removing cumulus cells from oocyte-cumulus complexes 20 h after the start of IVM improves the meiotic competence of oocytes derived from both SF and MF, without any reduction of developmental competence of the oocytes following parthenogenetical activation.

263 REGULATION OF SEVERAL TRANSCRIPTS DURING IN VITRO MATURATION IN PORCINE OOCYTES COLLECTED FROM DIFFERENT SIZE FOLLICLES

2011 ◽  
Vol 23 (1) ◽  
pp. 229
Author(s):  
M. J. Izquierdo-Rico ◽  
R. Romar ◽  
C. Kohata ◽  
H. Funahashi
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Germinal Vesicle ◽  
Developmental Competence ◽  
Medium Size ◽  
Total Rna ◽  
Follicle Diameter ◽  
First Polar Body

Oocyte-specific transcripts play important roles in oocyte maturation, fertilization, and early embryonic development. Currently, oocytes from medium-size follicles have been used for different assisted reproductive techniques after in vitro maturation (IVM). The aim of this study was to compare the mRNA expression level in porcine oocytes collected from medium (3–6 mm) and small (<2 mm) size follicles. Genes were selected based on their described maternal effect (NALP9, HSF1), their identification as markers of oocyte maturation (AURK-A, AURK-B, MOS, and C-mos), their involvement in fertilization (ZP3, ZP4), and anti-apoptotic effect (Bcl-2). All transcripts were studied in oocytes just after collection [germinal vesicle (GV) stage] and after in vitro maturation (IVM; metaphase II stage). To ensure nuclear stage of immature oocytes, oocytes were mechanically denuded just after collection, centrifuged (10 000 rpm, 5 min, RT), and observed under the microscope (60×). Those oocytes with clear nucleolus and evident nuclear membrane were selected and stored (n = 10) until study. For metaphase II oocytes, only those exhibiting the extrusion of first polar body after IVM (n = 10) were selected. Total RNA was extracted from the pool of 10 immature and mature oocytes. One picogram of luciferase mRNA per oocyte was added as an exogenous standard. Total RNA was extracted from oocytes and cDNA was obtained and used as a template for quantitative PCR to analyse the level of different transcripts. The whole process was replicated 4 times. Data were normalized to the luciferase RNA and analysed by one-way ANOVA with maturational stage (GV or metaphase II) and follicle size (small or medium) as fixed factors. Results show that all transcripts were significantly decreased during IVM (P < 0.05). Therefore, after IVM, NALP9, AURK-A, MOS, C-mos, ZP3, ZP4, and Bcl-2 transcripts were significantly reduced in matured oocytes compared with immature ones irrespective of follicle diameter. Transcripts of AURKAB and HSF1 decreased after IVM in oocytes from medium follicles or small follicles, respectively. A significant effect of follicular size was only detected in MOS transcripts in GV-stage oocytes because those collected from middle follicles had a higher amount than the ones from small follicles (Table 1). These results suggest that the variations in the maternal store of RNA during IVM are not related with follicle diameter for the studied genes. Further investigations are necessary to determinate the developmental competence of oocytes that came from different types of follicles (small and medium follicles). Table 1.Variation of transcripts during in vitro maturation in porcine oocytes collected from small and medium follicles This study was supported by Okayama University. R. Romar was given a grant by JSPS (Ref. S-09210).

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 Effect of Trans-^|^epsilon;-Viniferin on In Vitro Porcine Oocyte Maturation and Subsequent Developmental Competence in Preimplantation Embryos

2013 ◽  
Vol 75 (10) ◽  
pp. 1277-1286 ◽  
Author(s):  
Yubyeol JEON ◽  
Seong-Sung KWAK ◽  
Seung-A CHEONG ◽  
Yeon Hee SEONG ◽  
Sang-Hwan HYUN
Keyword(s):  
Oocyte Maturation ◽  
Developmental Competence ◽  
Preimplantation Embryos ◽  
Porcine Oocyte

scholarly journals Iron Overload-Induced Ferroptosis Impairs Porcine Oocyte Maturation and Subsequent Embryonic Developmental Competence in vitro

2021 ◽  
Vol 9 ◽  
Author(s):  
Weiyi Hu ◽  
Yan Zhang ◽  
Dali Wang ◽  
Tingting Yang ◽  
Jiajia Qi ◽  
...  
Keyword(s):  
Cell Death ◽  
Iron Overload ◽  
Mitochondrial Dysfunction ◽  
Oocyte Maturation ◽  
Oocyte Quality ◽  
Developmental Competence ◽  
Ammonium Citrate ◽  
Regulated Cell Death ◽  
Porcine Oocyte

Accumulating evidence indicates that ferroptosis is an iron-dependent form of regulated cell death. This type of iron-dependent programmed cell death is different from traditional forms of regulated cell death, such as apoptosis and autophagy. However, the role of ferroptosis in porcine oocyte maturation and the associated mechanism remain unclear. In the present research, we investigated the effects of ferric ammonium citrate (FAC), a specific ferroptosis inducer, on porcine oocyte meiotic maturation and quality and subsequent embryonic developmental competence. FAC treatment caused obvious accumulation of intracellular ferrous ions in porcine oocytes. At the end of the in vitro maturation (IVM) period, there was a significant decrease in the polar body (PB) extrusion rate and an increase in the percentage of abnormal oocytes in the FAC treatment groups, indicating that iron overload-induced ferroptosis may suppress the meiotic process during porcine oocyte maturation. We also found that after FAC treatment, the subsequent two-cell rate, four-cell rate and blastocyst formation rate were significantly decreased in porcine parthenogenetic activation (PA) embryos, indicating that iron overload-induced ferroptosis decreased porcine oocyte quality. Further analysis revealed that FAC treatment not only enhanced intracellular reactive oxygen species (ROS) generation, decreased intracellular free thiol levels and induced mitochondrial dysfunction but also triggered autophagy in porcine oocytes. Taken together, these findings suggest that iron overload-induced ferroptosis impairs porcine oocyte meiosis and decreases porcine oocyte quality, possibly by increasing oxidative stress, inducing mitochondrial dysfunction and triggering autophagy.

scholarly journals 55 NUCLEUS CHANGES AND DEVELOPMENT OF PORCINE RECONSTRUCTED (NT) AND PARTHOGENETICALLY ACTIVATED (PA) EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 177
Author(s):  
N.R. Mtango ◽  
T. Kono
Keyword(s):  
Polar Body ◽  
Cumulus Cells ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Normal Embryo ◽  
Basic Medium ◽  
Cell Nuclei ◽  
First Polar Body ◽  
Porcine Oocyte

Nuclear reprogramming is characterized by functional modification(s) of the transferred nucleus that allows it to direct normal embryo development with the potential to grow to term. The aim of our study was to investigate the process of nuclear changes in reconstructed and activated embryos as well as their developmental competence. All chemicals used were from Sigma Chemicals (St. Louis, MO, USA). Cumulus-oocyte complexes were aspirated from slaughterhouse ovaries of prepurbetal gilts and matured for 42 h in vitro. The cumulus cells were removed by adding in 1 mg mL −1 hyaluronidase in TLP-HEPES. For the NT experiment, oocytes with first polar body were cultured in 0.4 μg mL−1 demecolcine for 1 h. A protruding membrane was removed by micromanipulator and a single donor nucleus from fetal fibroblast was injected subzonally. Fusion was conducted immediately after transfer in 0.3 M mannitol, 0.5 mM HEPES, 0.1% PVA, and 0.1 mM MgCl2 in a fusion chamber with parallel electrodes set 1 mm apart using a singe DC pulse of 125 V mm−1 for 80 s. Activation was done 2–4 h after fusion in the same medium as fusion but with 0.1 mM CaCl2 added; embryos were cultured in 5 μg mL−1 cytochalasin B and 10 μg mL−1 cyclohexamide for 6 h. The embryos were cultured in glucose-free NCSU-37 containing 4 mg mL−1 BSA as basic medium supplemented with 0.17 mM sodium pyruvate and 2.73 mM sodium lactate from Days 0 to 2, and then in basic medium with 5.55 mM D-glucose from Days 2–6 (Kikuchi K et al. 2002 Biol. Reprod. 66, 1033–1041). Non-manipulated oocytes (PA) were electrically activated as stated above. For observing the changes of donor cells, some reconstructed oocytes were fixed 2 h after fusion, prior to activation, and some 12 h after activation in acetic acid:ethanol (1:3) and stained in 1% orcein. The activated oocytes were fixed at 12 h and stained as stated above. There were 47.5% (38/80) of reconstructed oocytes with premature chromosome condensation (PCC), and 23.7% (19/80) with nuclear swelling two hours after fusion. Pronuclear like formation 12 h after activation was 45% (27/60) and 83.3% (50/60) in NT and PA, respectively. The blastocyst rate was 8.3% (5/60) and 46% (69/150) for NT and PA, respectively. The results suggest that porcine oocyte cytoplasm can successfully reprogram somatic cell nuclei and support the development of NT embryos to the blastocyst stage.

scholarly journals Rosmarinic acid treatment during porcine oocyte maturation attenuates oxidative stress and improves subsequent embryo development in vitro

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6930
Author(s):  
Yan Zhang ◽  
Jing Guo ◽  
Xiao Wei Nie ◽  
Zi Yue Li ◽  
Yu Meng Wang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oocyte Maturation ◽  
Rosmarinic Acid ◽  
Reactive Oxygen Species Generation ◽  
Subsequent Development ◽  
Developmental Competence ◽  
Control Group ◽  
Porcine Oocyte ◽  
Number Of Cells

Background In vitro maturation (IVM) of oocytes has been widely used in the field of assisted reproductive technology. However, oocytes can be injured by oxidative stress during the process of IVM. Methods The present study was designed to evaluate the influences of rosmarinic acid (RA) on the IVM of porcine oocytes and the subsequent development of early-stage embryos as well as its underlying mechanisms. Various concentrations of RA (5 µM, 10 µM, and 25 µM) were treated with porcine oocyte maturation medium during the period of IVM. Results and Discussion The results showed that 5 µM RA treatment during the period of porcine oocyte IVM improves blastocyst quality and hatching ability after parthenogenetic activation. Furthermore, the presence of RA during the period of IVM dramatically improved the total number of cells after somatic cell nuclear transfer compared to the number of cells in the control group. Notably, RA treatment during the period of porcine oocyte IVM decreased intracellular reactive oxygen species generation not only in oocytes but also in cumulus cells. Further analysis showed that the intracellular free thiols levels in the oocytes were enhanced by treatment with RA during the period of porcine oocyte IVM compared to the free thiols levels in the control groups. These results indicate that RA improves the developmental competence of porcine oocytes during the IVM period by attenuating oxidative stress.

scholarly journals Iron Overload Is Associated With Accelerated Progression of Osteoarthritis: The Role of DMT1 Mediated Iron Homeostasis

2021 ◽  
Vol 8 ◽  
Author(s):  
Xingzhi Jing ◽  
Jiamin Lin ◽  
Ting Du ◽  
Zhensong Jiang ◽  
Tao Li ◽  
...  
Keyword(s):  
Iron Overload ◽  
Inflammatory Cytokines ◽  
Iron Homeostasis ◽  
Cartilage Destruction ◽  
Mice Model ◽  
Cellular Iron ◽  
Cellular Iron Homeostasis ◽  
Pro Inflammatory Cytokines

Objective: Iron overload is common in elderly people which is associated with an increased prevalence of osteoarthritis (OA), but the exact role of iron in the development of OA has not been established. The aim of the present study is to elucidate the connection between iron overload and OA using an iron overloaded mice model, as well as to explore the role of iron homeostasis, iron transporters dependent iron influx in OA pathogenesis.Methods: The iron overloaded mice model was established and OA was surgically induced. OA progression was assessed at 8 weeks after surgery. Next, primary chondrocytes were treated with pro-inflammatory cytokines and iron regulators mediated iron homeostasis were evaluated. Involvement of iron transporters was analyzed using chondrocytes mimicking an osteoarthritis-related phenotype in vitro.Results: Iron overloaded mice exhibited greater cartilage destruction and elevated ADAMTS5 as well as MMP13 expression along with increased iron accumulation and dysregulated iron regulators. Pro-inflammatory cytokines could disturb cellular iron homeostasis via upregulating iron import proteins, TFR1 and DMT1, downregulating iron efflux protein FPN, thus result in cellular iron overload. Among iron transporters, DMT1 was found to play pivotal roles in iron overload induced OA progress. Inhibition of DMT1 suppressed IL-1β induced inflammatory response and ECM degradation via blockade of MAPK and PI3K/AKT/NF-κB pathways.Conclusions: Our results suggest that iron takes parts in the development of OA and cutting iron influx via inhibiting DMT1 activity could be an attractive new target for OA treatment.

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