Mogroside V Protects Porcine Oocytes From Lipopolysaccharide-Induced Meiotic Defects

Accumulating evidence has demonstrated that lipopolysaccharide (LPS) compromises female reproduction, especially oocyte maturation and competence. However, methods to protect oocyte quality from LPS-induced deterioration remain largely unexplored. We previously found that mogroside V (MV) can promote oocyte maturation and embryonic development. However, whether MV can alleviate the adverse effects of LPS exposure on oocyte maturation is unclear. Thus, in this study, we used porcine oocytes as a model to explore the effects of MV administration on LPS-induced oocyte meiotic defects. Our findings show that supplementation with MV protected oocytes from the LPS-mediated reduction in the meiotic maturation rate and the subsequent blastocyst formation rate. In addition, MV alleviated the abnormalities in spindle formation and chromosome alignment, decrease in α-tubulin acetylation levels, the disruption of actin polymerization, and the reductions in mitochondrial contents and lipid droplet contents caused by LPS exposure. Meanwhile, LPS reduced m6A levels in oocytes, but MV restored these epigenetic modifications. Furthermore, MV reduced reactive oxygen species (ROS) levels and early apoptosis in oocytes exposed to LPS. In summary, our study demonstrates that MV can protect oocytes from LPS-induced meiotic defects in part by reducing oxidative stress and maintaining m6A levels.

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Mogroside V Alleviates Oocyte Meiotic Defects and Quality Deterioration in Benzo(a)pyrene-Exposed Mice

Frontiers in Pharmacology ◽

10.3389/fphar.2021.722779 ◽

2021 ◽

Vol 12 ◽

Author(s):

Lumin Sui ◽

Ke Yan ◽

Huiting Zhang ◽

Junyu Nie ◽

Xiaogan Yang ◽

...

Keyword(s):

Actin Polymerization ◽

Formation Rate ◽

Female Reproduction ◽

Acetylated Tubulin ◽

Quality Deterioration ◽

Bioactive Component ◽

Intracellular Ros ◽

Chromosome Alignment ◽

Maturation Rate ◽

Oocyte Meiotic Maturation

Accumulating evidence has demonstrated that benzo(a)pyrene (BaP) exposure adversely affects female reproduction, especially oocyte meiotic maturation and subsequent embryo development. Although we previously found that mogroside V (MV), a major bioactive component of S. grosvenorii, can protect oocytes from quality deterioration caused by certain stresses, whether MV can alleviate BaP exposure-mediated oocyte meiotic defects remains unknown. In this study, female mice were exposed to BaP and treated concomitantly with MV by gavage. We found that BaP exposure reduced the oocyte maturation rate and blastocyst formation rate, which was associated with increased abnormalities in spindle formation and chromosome alignment, reduced acetylated tubulin levels, damaged actin polymerization and reduced Juno levels, indicating that BaP exposure results in oocyte nucleic and cytoplasmic damage. Interestingly, MV treatment significantly alleviated all the BaP exposure-mediated defects mentioned above, indicating that MV can protect oocytes from BaP exposure-mediated nucleic and cytoplasmic damage. Additionally, BaP exposure increased intracellular ROS levels, meanwhile induced DNA damage and early apoptosis in oocytes, but MV treatment ameliorated these defective parameters, therefore it is possible that MV restored BaP-mediated oocyte defects by reducing oxidative stress. In summary, our findings demonstrate that MV might alleviate oocyte meiotic defects and quality deterioration in BaP-exposed mice.

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RBM14 Modulates Tubulin Acetylation and Regulates Spindle Morphology During Meiotic Maturation in Mouse Oocytes

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.635728 ◽

2021 ◽

Vol 9 ◽

Author(s):

Hao Qin ◽

Yi Qu ◽

Yi-Feng Yuan ◽

Yang-Yang Li ◽

Jie Qiao

Keyword(s):

Oocyte Maturation ◽

Mammalian Cells ◽

Rna Binding ◽

Oocyte Quality ◽

Meiotic Maturation ◽

Tubulin Acetylation ◽

Potential Biomarker ◽

Chromosome Alignment ◽

Oocyte Meiotic Maturation ◽

Old Mice

RBM14 is an RNA-binding protein that regulates spindle integrity in mitosis; however, its functions during meiosis are still unclear. In this study, we discovered that RBM14 expression was down-regulated in oocytes from old mice. The RBM14 distribution at different stages of meiosis was explored, while it presents overlapped localization patterns with α-tubulin in MI- and MII-stage oocytes. Treatment of MI-stage oocytes with spindle-perturbing agents revealed that RBM14 was co-localized with microtubules. RBM14 knockdown with RBM14-specific morpholino showed that RBM14-depleted oocytes underwent symmetric division compared to the controls. RBM14 knockdown also resulted in spindle defects and chromosome abnormalities during oocyte maturation, presumably due to α-tubulin hyperacetylation. Co-immunoprecipitation analysis demonstrated that RBM14 is interacted with endogenous α-tubulin in mammalian cells. These findings indicate that RBM14 is an essential modulator of oocyte meiotic maturation by regulating α-tubulin acetylation to affect spindle morphology and chromosome alignment. Consequently, RBM14 represents a potential biomarker of oocyte quality and a novel therapeutic target in women with oocyte maturation failure.

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FKBP25 Regulates Meiotic Apparatus During Mouse Oocyte Maturation

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.625805 ◽

2021 ◽

Vol 9 ◽

Author(s):

Danni Wang ◽

Hongzheng Sun ◽

Jiaqi Zhang ◽

Zhenyue Huang ◽

Congyang Li ◽

...

Keyword(s):

Oocyte Maturation ◽

Ribosome Biogenesis ◽

Phosphorylation Site ◽

Ectopic Expression ◽

Oocyte Quality ◽

Female Reproduction ◽

Microtubule Stability ◽

Microtubule Attachment ◽

Fk506 Binding Proteins ◽

Maternal Aging

FK506 binding proteins 25 (FKBP25) has been shown to function in ribosome biogenesis, chromatin organization, and microtubule stability in mitosis. However, the role of FKBP25 in oocyte maturation has not been investigated. Here, we report that oocytes with FKBP25 depletion display abnormal spindle assembly and chromosomes alignment, with defective kinetochore-microtubule attachment. Consistent with this finding, aneuploidy incidence is also elevated in oocytes depleted of FKBP25. Importantly, FKBP25 protein level in old oocytes is significantly reduced, and ectopic expression of FKBP25 could partly rescue the aging-associated meiotic defects. In addition, by employing site-specific mutagenesis, we identify that serine 163 is a major, if not unique, phosphorylation site modulating the action of FKBP25 on meiotic maturation. In summary, our data indicate that FKBP25 is a pivotal factor for determining oocyte quality, and may mediate the effects of maternal aging on female reproduction.

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307 MATURATION RATE AND GENE EXPRESSION ANALYSIS OF GOAT OOCYTES SELECTED BY FOLLICLE SIZE AND BRILLIANT CRESYL BLUE STAINING

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab307 ◽

2015 ◽

Vol 27 (1) ◽

pp. 242

Author(s):

M. Yang ◽

S. Hu ◽

L. Cox ◽

M. Regouski ◽

H. Rutigliano ◽

...

Keyword(s):

Gene Expression ◽

Oocyte Maturation ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Oocyte Quality ◽

Developmental Competence ◽

Brilliant Cresyl Blue ◽

Relative Expression ◽

Cresyl Blue ◽

Maturation Rate

Oocyte quality plays a critical role in determining the success of embryo development. Studies on cattle and goats indicate that oocytes derived from large follicles (LFO) have greater developmental competence than those derived from small follicles (SFO). Brilliant cresyl blue (BCB) staining determines the activity of glucose-6-phosphate dehydrogenase and is a commonly used noninvasive marker of oocyte competence. Studies in pigs, goats, cows, mice, and dogs showed that the maturation and blastocyst developmental rate of BCB+ oocytes is significantly higher than BCB– oocytes. The aim of this study was to evaluate the maturation rate of goat oocytes selected based on follicular size and BCB staining and compare their relative patterns of gene expression. Maturation rate and gene expression profile were expected to be different in these oocyte groups. Cumulus-oocyte complexes were recovered from abattoir-derived ovaries using a slicing technique. Eleven rounds of oocyte maturation and 4 rounds of BCB staining were carried out. During each replicate, oocytes from large (≥3 mm) and small (<3 mm) follicles were collected separately from the same group of ovaries. Oocyte maturation rates were 54.3 ± 5.4% for LFO (n = 378) and only 33.5 ± 3.7% for SFO (n = 981; P < 0.01). The BCB+ (n = 223) oocytes yielded a significantly higher maturation rate than the BCB– (n = 194) oocytes (56.1 ± 1.8 v. 20.6 ± 3.8%, respectively; P < 0.001). Gene expression analysis was conducted on individual MII oocytes (21 oocytes per group). Specific target amplification was performed on a single oocyte directly by using the CellsDirect One-Step qRT–PCR Kit (Invitrogen). Quantitative real-time PCR was then performed using the 48.48 BioMark platform from Fluidigm. Forty two genes were selected from the following categories: growth factors, transcription factors, metabolism, pluripotency, cell cycle, apoptosis, and oocyte-specific genes. Relative expression values were calculated using the ΔΔCT (fold change) method and analysed by ANOVA. The significance was assigned at P < 0.05. The relative expression of CCNA2, CDK2, CCNB1, POU5F1, SOX2, EGF, FGF2, GDF9, ZP3, BCL2, GJA1, DDR1, PFKFB3, IGF2R, and GRB10 was significantly greater (P < 0.05) in both LFO and BCB+ oocytes compared to SFO and BCB– oocytes, respectively. The proapoptotic gene BAX, the ACSL3 gene involved in fatty acid oxidation, and the growth factor IGF1 were expressed significantly higher (P < 0.05) in SFO compared to LFO. By investigating these differentially expressed transcripts, we will better understand pathways involved in oocyte developmental competence and potentially use them as markers of oocyte quality. We expect that the ability to select oocytes of better quality based on BCB staining will improve outcomes of IVF and SCNT.

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P–434 Effect of let–7a mimic as a new pharmaco-protective agent against chemotherapy-induced ovarian damage on subsequent follicular development and oocyte quality using mouse ovarian transplantation model

Human Reproduction ◽

10.1093/humrep/deab130.433 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

C Alexandri ◽

A Nguyen ◽

G Va. De. Steen ◽

I Demeestere

Keyword(s):

Fertility Preservation ◽

Oocyte Maturation ◽

Follicular Development ◽

Oocyte Quality ◽

Adult Mice ◽

Maturation Rate ◽

Ovarian Damage ◽

The Impact

Abstract Study question What is the long-term impact of let–7a-mimic transfection on oocytes development in new-born mice ovaries exposed to chemotherapy in vitro following transplantation in the kidney? Summary answer The let–7a-mimic restoration protects against chemotherapy-induced ovarian apoptosis and preserves subsequent follicular developmental and acquisition of oocyte maturation competence in mouse. What is known already It is well known that cyclophosphamide and its active metabolites (4-hydroperoxycyclophosphamide, 4-HC) cause irreversible ovarian damage and impair future fertility of cancer survivors. Besides the available fertility preservation options, microRNAs/miRNAs appear to be very attractive and novel targets to prevent theses damage. We showed that miRNAs were dysregulated after exposure to 4-HC in postnatal-day–3 (PND3) ovaries, let–7a being the most downregulated among them. By replacing let–7a function, let–7a-mimic was able to protect mouse follicles against 4-HC in vitro. This previous study suggested that it could preserve the reproductive potential after treatment. However, the impact on subsequent oocytes development is unknown Study design, size, duration PND3 ovaries from C57blxCBAF1 hybrid mice were cultured under 3 conditions: control, chemotherapy for 24h (4-HC/20μΜ/24h), chemotherapy for 24h+let–7a-mimic (4-HC/20μΜ/24h+let–7a-mimic). Nine PND3 ovaries were cultured in the different conditions and then transplanted under the kidney’s capsule of C57blxCBAF1 hybrid adult mice for follicular growth/apoptosis evaluation. Then, 21 ovaries (≥7/condition) were used for oocyte maturation assessment after transplantation and ovarian stimulation. All transplanted mice were observed during 21 days before PND3 ovaries collection. Participants/materials, setting, methods: PND3 ovaries were cultured in vitro using inserts under different conditions. A liposome-based system was used to deliver let–7a-mimic into ovaries and QPCR-assays validated its expression levels after transfection. Apoptosis was evaluated by TUNEL Assay while haematoxylin/eosin staining was used for assessing the follicular morphology, stage and count. The oocyte maturation rate was evaluated at day 21 post-transplantation after gonadotropins injection, mechanical eggs collection and in vitro maturation for 24 hours. Main results and the role of chance The apoptosis assessment confirmed that let–7a-mimic transfection reduced the chemotherapy-induced damage in PND3 ovaries in vitro. The number of primordial follicles was significantly reduced (p < 0.05) compared to control after chemotherapy exposure. However, it was increased in chemo24h+let–7a-mimic compared to chemo24h alone while remaining lower than control (p > 0.05). Accordingly, the number of the transitory follicles reflecting follicular activation was significantly higher in chemo24h compared to control (p < 0.05) and chemo24h+let–7a-mimic but for the last one, the result was not significant. Consequently, chemotherapy induces follicle activation while let–7a restoration tends to slow down this effect. To evaluate the long-term effects of chemotherapy and let–7a-mimic transfection, in vitro exposed PND3 ovaries were transplanted under kidney’s capsule in female adult mice. After 21 days, the ovarian reserve was higher in control, but we observed a slight increase of follicular density in the chemo24+let–7a-mimic compared to chemo24h. Similarly, the percentage of damaged/apoptotic cells was higher in all chemotherapy exposed groups compared to control but the impact was lower after let–7a restoration (12,0% and 28,2% in chemo24+let–7a-mimic and chemo24h, respectively). Importantly, the oocyte maturation rate after transplantation was higher in chemo24h+let–7a-mimic compared to chemo24h (40% versus 18%, respectively), suggesting a preservation of oocytes maturation competence. Limitations, reasons for caution The multiple in vitro/in vivo steps may introduce study bias. Moreover, the oocyte competence and live offspring is currently evaluated. The blastocyst formation and embryo development from oocytes fertilized in vitro, are more relevant parameters for oocyte quality assessment. The birth of healthy animals will confirm let–7a-mimic-transfection safety. Wider implications of the findings: Our previous study demonstrated the anti-apoptotic effect of let–7a restoration in mouse ovaries against chemotherapy. In the current study, we demonstrated a long-term beneficial effect of let–7a restoration strategy on follicular development and oocytes maturation capacity. The results open new perspectives in fertility preservation using pharmacological approach. Trial registration number Not applicable

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Effects of the Oocyte Maturation Rate on Clinical Outcomes of Conventional IVF and ICSI in Female Patients Aged ≥38 Years

10.21203/rs.3.rs-80046/v1 ◽

2020 ◽

Author(s):

shuang liu ◽

Hongjun Yu ◽

Baoshan Li ◽

Chunyi Li ◽

Dongkai Cheng

Keyword(s):

Pregnancy Rate ◽

Oocyte Maturation ◽

Birth Rate ◽

Formation Rate ◽

Live Birth Rate ◽

Fertilization Rate ◽

Blastocyst Formation ◽

Oocyte Maturity ◽

Cleavage Rate ◽

Maturation Rate

Abstract Research Question: The average female reproductive potential peaks at age 25 and then begins to decline. As women are delaying childbearing, the prevalence of infertility has risen, leading to increasing demand for assisted reproductive technology (ART). Oocyte quality remains the most important issue during in vitro fertilization (IVF) cycles. This study investigated the effects of the oocyte maturation rate (OMR) on clinical outcomes of conventional IVF and intracytoplasmic sperm injection (ICSI) in female patients aged ≥38 years. Design: A retrospective analysis of 6562 infertile patients who were treated with IVF/ICSI at the reproductive medicine center of our hospital from January 2011 to December 2017 was performed. According to the oocyte maturity (the ratio of the number of mature oocytes to the number of oocytes) on the day of egg collection, the patients were divided into three groups: group A (oocyte maturity ≤30%, n=422), and group B (oocyte maturity from 30-75%, n=1290), and group C (oocyte maturity ≥75%, n=4850). The patient age, years of infertility years, days of gonadotropin (Gn), Gn dosage, serum luteinizing hormone (LH), estradiol (E2), and progesterone (P) levels on the day of human chorionic Gn (HCG) injection, E2 levels per mature oocyte, E2 levels per oocyte, number of mature oocytes, oocyte recovery rate, fertilization rate, cleavage rate, excellent embryo rate, blastocyst formation rate, pregnancy rate, and live birth rate were compared among the three groups. Results: Factors including age, years of infertility, number of eggs obtained, number of mature eggs, normal fertilization rate, cleavage rate, excellent embryo rate, blastocyst formation rate, pregnancy rate, and live birth rate were all found to be related to oocyte maturity on the day of HCG administration (P<0.05). Conclusions: There is a close relationship between oocyte maturity and embryo quality, and a low OMR may be related to a poor ovarian response or decreased sensitivity to Gn. Therefore, a low OMR may affect the fertilization and embryonic development potential in elderly patients undergoing IVF/ICSI, thus affecting the pregnancy rate.

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In vitro maturation of porcine oocytes with retinoids improves embryonic development

Reproduction Fertility and Development ◽

10.1071/rd07175 ◽

2008 ◽

Vol 20 (4) ◽

pp. 483 ◽

Cited By ~ 19

Author(s):

C. Almiñana ◽

M. A. Gil ◽

C. Cuello ◽

I. Caballero ◽

J. Roca ◽

...

Keyword(s):

Embryo Development ◽

Oocyte Maturation ◽

In Vitro Maturation ◽

Formation Rate ◽

Control Treatment ◽

In Vitro Fertilisation ◽

Blastocyst Formation ◽

Maturation Rate ◽

And Control

In the present study, the effects of retinoid metabolite administration during in vitro maturation (IVM) on oocyte maturation, parameters of in vitro fertilisation (IVF) and embryo development were examined. Varying concentrations of 9-cis retinoic acid (RA; 0, 5, 50 and 500 nm; Experiment 1) and all-trans retinol (ROH; 0, 125, 1250 and 12 500 nm; Experiment 2) were included in the maturation medium. Cumulus–oocyte complexes were matured in vitro and inseminated with frozen–thawed spermatozoa. Presumptive zygotes were cultured for 16 h to assess IVF parameters or for 7 days to assess embryo development and quality. In Experiment 1, the oocyte maturation rate to metaphase II was significantly decreased (P < 0.001), with values below 5%, in the presence of the highest concentration of RA (500 nm). However, 5 and 50 nm RA had no effect compared with control. Treatment with 5 nm RA improved the blastocyst development rate (P < 0.001). In Experiment 2, the oocyte maturation rate did not differ between 125 and 1250 nm ROH treatment groups and control. However, treatment with 12 500 nm ROH was deleterious because no matured oocytes were observed following the treatment. The penetration rate was lower in the group treated with 1250 nm ROH compared with the 125 nm ROH-treated and control groups, but the blastocyst formation rate did not differ among the three groups. In conclusion, 5 nm RA in the IVM medium significantly increased the blastocyst formation rate, suggesting that RA may play an important role during IVM.

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