The effect of FF-MAS on porcine cumulus–oocyte complex maturation, fertilization and pronucleus formation in vitro

Zygote ◽  
2006 ◽  
Vol 14 (3) ◽  
pp. 189-199 ◽  
Author(s):  
Inger Faerge ◽  
Frantisek Strejcek ◽  
Jozef Laurincik ◽  
Detlef Rath ◽  
Heiner Niemann ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Developmental Potential ◽  
Nuclear Maturation ◽  
Cumulus Oocyte Complex ◽  
Constant Ph ◽  
Porcine Oocyte ◽  
Fine Glass ◽  
Epidermal Growth

SummaryFollicular fluid meiosis-activating sterol (FF-MAS) has been isolated from the follicular fluid (FF) of several species including man. FF-MAS increases the quality of in vitro oocyte maturation, and thus the developmental potential of oocytes exposed to FF-MAS during in vitro maturation is improved. The aim of the present study was to investigate the effects of FF-MAS on porcine oocyte maturation and pronucleus formation in vitro. Porcine cumulus–oocyte complexes (COCs) were isolated from abattoir ovaries and in vitro matured for 48 h in NCSU 37 medium supplemented with 1 mg/l cysteine, 10 ng/ml epidermal growth factor and 50 μM 2-mercaptoethanol with or without 10% porcine follicular fluid (pFF). For the first 22 h, 1 mM db-cAMP and 10 I.E PMSG/hCG was added. The medium was supplemented with 1 μM, 3 μM, 10 μM, 30 μM or 100 μM FF-MAS dissolved in ethanol. After maturation the COCs were denuded mechanically using a fine glass pipette under constant pH and in vitro fertilized with fresh semen (5 × 105 spermatozoa/ml). The presumptive zygotes were evaluated 18 h after fertilization. The addition of pFF increased the monospermic as well as the polyspermic penetration of oocytes. In the absence of pFF, the addition of FF-MAS decreased the polyspermic penetration rate, whereas FF-MAS in combination with pFF decreased monospermic and increased polyspermic penetration. The degeneration rate of ova decreased in the presence of FF-MAS irrespective of the presence or absence of pFF. In the absence of pFF, FF-MAS at 3–10 μM increased the number of zygotes with advanced maternal pronuclear stages. In supraphysiological doses, i.e. 30–100 μM, FF-MAS dose-dependently and reversibly inhibited nuclear maturation in the absence of pFF.

Epidermal growth factor enhances porcine oocyte maturation in vitro in the absence of follicular fluid or hormones

1993 ◽  
Vol 39 (1) ◽  
pp. 180 ◽  
Author(s):  
J. Arellano ◽  
M.J. Illera ◽  
P. Lorenzo ◽  
J. Sanchez ◽  
G. Silván ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Epidermal Growth

scholarly journals Oocyte Competence Biomarkers Associated With Oocyte Maturation: A Review

2021 ◽  
Vol 9 ◽  
Author(s):  
Batara Sirait ◽  
Budi Wiweko ◽  
Ahmad Aulia Jusuf ◽  
Dein Iftitah ◽  
R. Muharam
Keyword(s):  
Oocyte Maturation ◽  
Blastocyst Stage ◽  
Current Approach ◽  
Developmental Competence ◽  
Matrix Remodeling ◽  
Oocyte Competence ◽  
Nuclear Maturation ◽  
Oocyte Developmental Competence ◽  
Cumulus Oocyte Complex

Oocyte developmental competence is one of the determining factors that influence the outcomes of an IVF cycle regarding the ability of a female gamete to reach maturation, be fertilized, and uphold an embryonic development up until the blastocyst stage. The current approach of assessing the competency of an oocyte is confined to an ambiguous and subjective oocyte morphological evaluation. Over the years, a myriad of biomarkers in the cumulus-oocyte-complex has been identified that could potentially function as molecular predictors for IVF program prognosis. This review aims to describe the predictive significance of several cumulus-oocyte complex (COC) biomarkers in evaluating oocyte developmental competence. A total of eight acclaimed cumulus biomarkers are examined in the study. RT-PCR and microarray analysis were extensively used to assess the significance of these biomarkers in foreseeing oocyte developmental competence. Notably, these biomarkers regulate vital processes associated with oocyte maturation and were found to be differentially expressed in COC encapsulating oocytes of different maturity. The biomarkers were reviewed according to the respective oocyte maturation events namely: nuclear maturation, apoptosis, and extracellular matrix remodeling, and steroid metabolism. Although substantial in vitro evidence was presented to justify the potential use of cumulus biomarkers in predicting oocyte competency and IVF outcomes, the feasibility of assessing these biomarkers as an add-on prognostic procedure in IVF is still restricted due to study challenges.

208 EFFECT OF GDF8 AND SB-431542 ON PORCINE OOCYTE DURING IN VITRO MATURATION AND SUBSEQUENT EMBRYONIC DEVELOPMENT

2016 ◽  
Vol 28 (2) ◽  
pp. 235
Author(s):  
J. D. Yoon ◽  
E. Lee ◽  
S.-H. Hyun
Keyword(s):  
Treatment Group ◽  
Embryonic Development ◽  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Transforming Growth Factor ◽  
Formation Rate ◽  
Transforming Growth Factor Β ◽  
Nuclear Maturation ◽  
Porcine Oocyte

Growth differentiation factor-8 (GDF8) is a member of the transforming growth factor-β that has been identified as a strong physiological regulator. SB-431542 (SB) is a specific inhibitor of transforming growth factor-β superfamily type I activin receptor-like kinase (ALK) receptors such as ALK4, ALK5, and ALK7. The purpose of this study is investigation of the effects of GDF8 and SB on porcine oocytes during in vitro maturation and subsequent embryonic development. We first performed ELISA to detect GDF8 concentrations in follicular fluid for each size of follicle; sizes were as follows: small (<3 mm), medium (>3 mm and <6 mm), and large (>6 mm) follicle. After detection of the GDF8 concentration in follicular fluid, we investigated the effect of GDF8 and SB treatment during in vitro maturation (IVM) on nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels, and embryonic development after IVF and parthenogenetic activation (PA). Data were analysed by ANOVA followed by Duncan using SPSS (Statistical Package for Social Science, IBM, New York, NY, USA) mean ± SEM. The ELISA result showed different concentrations of GDF8 for each grade of follicular fluid: small, 0.479 ng mL–1; medium, 0.668 ng mL–1; and large, 1.318 ng mL–1. During the IVM process, 1.318 ng mL–1 of GDF8 and 5 ng mL–1 of SB were added to the maturation medium as control, SB, SB+GDF8, and GDF8 treatment groups. After 44 h of IVM, GDF8 group (90.4%) showed a significantly higher nuclear maturation rate than control and SB+GDF8 groups (85.4 and 81.7%). The SB group (78.9%) showed significantly reduced nuclear maturation rate compared with control (P < 0.05). The GDF8 treatment group showed a significant decreased intracellular ROS and increased GSH levels compared with other groups (P < 0.05). The SB+GBF8 treatment group showed a significantly better cytoplasmic maturation than the SB treatment group. In the PA embryonic development analysis, the GDF8 treatment group showed a significantly higher blastocyst formation rate compared with other groups (47.9, 37.2, 46.4, and 58.7% respectively; P < 0.05). In the IVF embryonic development analysis, the GDF8 treatment groups showed significantly higher blastocyst formation rate compared with the SB group (28.2 and 42.2%, respectively; P < 0.05). In conclusion, treatment with GDF8 during porcine oocyte IVM improved the embryonic developmental competence via increased cytoplasmic maturation and led to better oocyte maturation from the ALK receptor inhibition by SB.

scholarly journals Inhibition of in vitro maturation of equine oocytes by interleukin 1 beta via specific IL-1 receptors

Reproduction ◽  
2003 ◽  
pp. 509-515 ◽  
Author(s):  
A Martoriati ◽  
M Caillaud ◽  
G Goudet ◽  
N Gerard
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Interleukin 1 ◽  
Inhibitory Effect ◽  
Interleukin 1 Beta ◽  
Nuclear Maturation ◽  
Epidermal Growth ◽  
Dose Dependent

Interleukin 1 beta (IL-1 beta) inhibits the LH-induced resumption of meiosis of equine oocytes in vitro. The present study was performed to clarify this inhibitory effect of IL-1 beta by testing increasing concentrations of IL-1 beta, and by measuring the effect of addition of IL-1 receptor antagonist (IL-1RA) to the culture medium. The effect of IL-1 beta on epidermal growth factor (EGF)-induced resumption of meiosis was also studied. Cumulus-oocyte complexes (COCs) were collected from subordinate follicles on ovaries obtained from an abattoir. In five distinct experiments, COCs were cultured for 30 h and nuclear maturation of oocytes was evaluated by DNA staining. In Expt 1, seven different media were tested: medium 1 (TCM199+BSA); medium 2 (medium 1+50 ng IL-1 beta ml(-1)); medium 3 (medium 1+eLH); and media 4, 5, 6 and 7 (medium 3 containing 0.1, 1.0, 10.0 and 50.0 ng IL-1 beta ml(-1), respectively). In Expt 2, four different media were tested: medium 1 (TCM199+BSA+eLH); medium 2 (medium 1+50 ng IL-1 beta ml(-1)); and media 3 and 4 (medium 2+IL-1RA at 50 and 100 ng ml(-1), respectively). In Expt 3, three different media were tested: medium 1 (TCM199+BSA+eLH); medium 2 (medium 1+50 ng IL-1RA ml(-1)); and medium 3 (medium 2+50 ng IL-1 beta ml(-1)). In Expt 4, four different media were tested: medium 1 (TCM199+BSA+eLH); and media 2, 3 and 4 (medium 1+IL-1RA at 50, 100 and 150 ng ml(-1), respectively). In Expt 5, three different media were tested: medium 1 (TCM199+BSA+EGF); medium 2 (medium 1+50 ng IL-1 beta ml(-1)); and medium 3 (medium 2+50 ng IL-1RA ml(-1)). In Expt 1, LH alone induced an increase in the rate of in vitro maturation (IVM) of equine oocytes (P<0.05), whereas IL-1 beta alone did not have any effect compared with medium 1. IL-1 beta (50 ng ml(-1)) significantly inhibited the eLH-induced IVM of oocytes (P<0.05) compared with medium 3. A decrease in rate of maturation was observed from a concentration of 10 ng IL-1 beta ml(-1) onwards. In Expt 2, the presence of IL-1RA in the culture medium inhibited the effect of IL-1 beta and restored the rate of oocyte maturation (P<0.05) observed in the presence of LH alone. In Expts 3 and 4 it was demonstrated that IL-1RA alone had no positive effect on the eLH-induced rate of maturation. In Expt 5, IL-1 beta inhibited the EGF-induced resumption of meiosis (P<0.05). The addition of IL-1RA inhibited this effect and restored the rate of oocyte maturation (P<0.05) observed with EGF alone. In conclusion, the present data confirm the inhibitory effect of IL-1 beta on IVM of equine oocytes induced by eLH and demonstrate its inhibitory effect on EGF-induced oocyte maturation. The rate of maturation decreased in a dose-dependent way and the lowest rate of maturation was observed at 50 ng IL-1 beta ml(-1) (P<0.05). The use of IL-1RA inhibited these effects, demonstrating that the action of IL-1 beta is receptor-mediated. Moreover, the results clearly show that, in equine species, IL-1 beta is involved in the physiology of COCs by regulating resumption of meiosis.

scholarly journals Effect of Interleukin-7 on In Vitro Maturation of Porcine Cumulus-Oocyte Complexes and Subsequent Developmental Potential after Parthenogenetic Activation

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 741
Author(s):  
Dongjin Oh ◽  
Joohyeong Lee ◽  
Eunhye Kim ◽  
Seon-Ung Hwang ◽  
Junchul-David Yoon ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Control Group ◽  
Developmental Potential ◽  
Parthenogenetic Activation ◽  
Nuclear Maturation ◽  
Interleukin 7

Interleukin-7 (IL-7) is a cytokine essential for cell development, proliferation and survival. However, its role in oocyte maturation is largely unknown. To investigate the effects of IL-7 on the in vitro maturation (IVM) of porcine oocytes, we analyzed nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, and subsequent embryonic developmental competence after parthenogenetic activation (PA) under several concentrations of IL-7. After IVM, IL-7 treated groups showed significantly higher nuclear maturation and significantly decreased intracellular ROS levels compared with the control group. All IL-7 treatment groups exhibited significantly increased intracellular GSH levels compared with the control group. All oocytes matured with IL-7 treatment during IVM exhibited significantly higher cleavage and blastocyst formation rates after PA than the non-treatment group. Furthermore, significantly higher mRNA expression levels of developmental-related genes (PCNA, Filia, and NPM2) and antioxidant-related genes (GSR and PRDX1) were observed in the IL-7-supplemented oocytes than in the control group. IL-7-supplemented cumulus cells showed significantly higher mRNA expression of the anti-apoptotic gene BCL2L1 and mitochondria-related genes (TFAM and NOX4), and lower transcript levels of the apoptosis related-gene, Caspase3, than the control group. Collectively, the present study suggests that IL-7 supplementation during porcine IVM improves oocyte maturation and the developmental potential of porcine embryos after PA.

scholarly journals Research on miRNAs of exosomes’ role on porcine oocyte development by extracted from follicular fluid

2020 ◽  
Author(s):  
Junhe Hu ◽  
Jinyi Dong ◽  
Zhi Zeng ◽  
Juan Wu ◽  
Xiansheng Tan ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Transgenic Animals ◽  
Oocyte Development ◽  
Clustering Methods ◽  
Sequencing Data ◽  
Genome Database ◽  
Porcine Oocyte ◽  
Follicular Size

Abstract Follicular development is crucial to oocyte maturation in these special surrounds, and the follicular size closely related to oocyte maturation. In order to clarify porcine oocyte maturation molecular mechanism, obtained exosomes miRNAs from porcine follicular fluid (PFF) are sequenced and researched among different follicular size as described in the methods. The results firstly show that PFF exosomes can be successfully isolated, and almost all valid reads of sequencing data of PFF exosomes can be successfully mapped to the porcine genome database. Using hierarchical clustering methods, it secondly finds that significantly expressed miRNAs can also be clustered in A, B C and D groups in heatmap according to different size follicles, which possibly to target mRNAs genes related to porcine oocyte development. Thirdly, through choosing ten significantly expressed miRNAs to predict the targeting genes for further GO analysis, the results show that the expression of neurotransmitter secretion genes changes greatly, and many targeting genes involves in regulation of FSH secretion, which is very closely related to oocyte maturation in growing follicles. Further, taking Pathway analysis for these targeting genes this ten choosing miRNAs, the results show that the pathways mainly related to the biosynthesis pathway of TGF-beta signaling pathway, which is very closely related to reproductive system function. Finally, these miRNAs in PFF EXs may provide a valuable addition about the mechanism of porcine oocyte maturation, which could be chosen as some molecular biomarkers to choose the high-quality oocytes for further porcine embryo production in vitro or transgenic animals research in the future.

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.

207 GANGLIOSIDE IMPROVES THE MEIOTIC MATURATION AND ANTI-APOPTOTIC EFFECTS DURING IN VITRO MATURATION OF PORCINE OOCYTES

2016 ◽  
Vol 28 (2) ◽  
pp. 234
Author(s):  
J.-W. Kim ◽  
H.-J. Park ◽  
S.-K. Chae ◽  
J.-H. Ahn ◽  
G.-Y. Do ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Egf Receptor ◽  
Untreated Group ◽  
Meiotic Maturation ◽  
Signal Molecules ◽  
Developmental Potential ◽  
Protein Levels ◽  
Porcine Oocyte ◽  
Apoptotic Effects

Gangliosides are sialic-acid-glycosphingolipids expressed in almost all vertebrate cells. These play a role in regulating cell differentiation and signal molecules of cellular signaling, including interactions with molecules of signal transduction pathways, such as receptors tyrosine kinases (RTK). Gangliosides perform critical functions through interactions with proteins as receptors in cell‐cell recognition, such as the epidermal growth factor (EGF) receptor (EGFR). Recent studies investigated functional a link between gangliosides-derived RTK signaling and embryo development until blastocyst in mice. However, there is no report on the directly relationship of GM3 or GD1a and oocyte maturation in pigs. Our studies have verified an interaction mechanism between EGF receptor signaling and gangliosides (GM3 and/or GD1a) in porcine oocyte during meiotic maturation and further development. The GM3 expression increased in mouse oocytes that were in vitro-matured (IVM) under apoptotic conditions. To induce the apoptotic conditions of oocytes, we used H2O2. We first confirmed that the expression of GM3 in oocytes of H2O2-treated groups (1 mM) was gradually increased compared with those of the control group (P < 0.05). We investigated the anti-apoptotic effect of GM3 as the inhibition apoptosis in porcine oocyte IVM by treatment of GM3 (5 or 10 μM) after pretreatment with H2O2. Next EGFR protein levels and meiotic maturation were investigated by Western blot analysis and acetic-orcerin staining, respectively, of IVM oocytes. The proportion of germinal vesicle arrested oocytes at 22 h was significantly increased (P < 0.05; 41.6 ± 1.5% v. 25.0 ± 0.0% in the untreated group) in EGF (10 ng mL–1) + GD1a (10 μM) treated group. To confirm the completion of meiotic maturation, we investigated the proportion of metaphase II (MII) oocytes at 44 h. The MII oocytes were increased (P < 0.05; 89.9 ± 3.6% v. 57.4 ± 5.3% in the untreated group) in oocytes cultured with EGF+GD1a. Interestingly, p-EGFR protein levels of matured oocytes were significantly increased (P < 0.05) by the EGF+GD1a treatment in the maturation process. In addition, EGF+GD1a treatment increased the proportion of normal pronucleus formation (2PN of 43.1 ± 5.2%) and increased (P < 0.05; 50.4 ± 6.1% v. 27.2 ± 2.7% in the untreated group) pre-implantation developmental potential until blastocyst. These results suggest that EGF+GD1a treatment improved the developmental competence of embryos via enhanced meiotic maturation of porcine oocytes by inducing EGFR activation. We confirmed that GM3 has anti-apoptotic effects during porcine oocyte maturation periods. Furthermore, these data will be helpful for better understanding the receptor mechanisms during oocyte maturation in pigs.

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