scholarly journals mRNA Expression and Role of PPARγ and PPARδ in Bovine Preimplantation Embryos Depending on the Quality and Developmental Stage

Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2358
Author(s):  
Katarzyna Suwik ◽  
Emilia Sinderewicz ◽  
Dorota Boruszewska ◽  
Ilona Kowalczyk-Zięba ◽  
Joanna Staszkiewicz-Chodor ◽  
...  
Keyword(s):  
Early Development ◽  
Embryo Development ◽  
Embryo Quality ◽  
Early Embryo ◽  
Developmental Competence ◽  
Preimplantation Embryos ◽  
Peroxisome Proliferator ◽  
Early Embryo Development ◽  
Quality Markers

Peroxisome proliferator-activated receptors (PPARs), a nuclear receptors for prostacyclin (PGI2) have been recognized as being essential for early embryo development. The objectives of the present study were to determine if the bovine early- and late-cleaved embryos in different stages of early development express PPARγ and PPARδ. Since embryo developmental competence depends on numerous biological factors, we evaluated if the expression of PPARγ and PPARδ correlate with selected embryo quality markers (SOX2, OCT4, PLAC8, IGF1R) in the in vitro produced embryos at different stages of their development. Developmental rates and embryo quality for early- and late-cleaved embryos were provided according to International Embryo Transfer Society (IETS; developmental stages: 2-, 4-, 16-cell embryo, morula, blastocyst (1—early, 2—developing, 3—expanded, 4—hatched); quality stages: A—high quality, B—moderate quality, C—low quality). We found that bovine embryos expressed mRNA of PPARδ and PPARγ at all stages of early development, independently of their quality. In addition, the expression of PPARδ and PPARγ correlated with the expression of quality markers in bovine blastocysts. Positive correlations were stronger and more frequent in the group of early-cleaved embryos, whereas the negative correlations were typical for the group of late-cleaved embryos. Obtained results and available literature reports may indicate the participation of PGI2, via PPARδ and PPARγ, in the processes related to the early embryo development, through the participation of this factor in the modulation of blastocyst hatching, implantation, and post-implantation development.

112 EFFECTS OF GUAIAZULENE ON IN VITRO CULTURE OF BOVINE ZYGOTES, AND ON mRNA TRANSCRIPTS RELATED TO EMBRYO QUALITY

2009 ◽  
Vol 21 (1) ◽  
pp. 156
Author(s):  
E. Dovolou ◽  
M. Clemente ◽  
G. S. Amiridis ◽  
I. Messinis ◽  
A. Kalitsaris ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Early Embryo ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Early Embryo Development ◽  
Mrna Transcripts ◽  
Oviductal Fluid ◽  
Maximum Humidity

We have previously shown that follicular and oviductal fluid provide greater total protection against lipid peroxidation than the respective media used for the in vitro embryo production. Reactive oxygen species (ROS) production has been implicated as a major cause for the reduced in vitro bovine embryo production; it is believed that they participate in meiotic arrest of oocytes, embryonic block and cell death. The aim of this study was to determine whether guaiazulene (G), an exogenous antioxidant, added in the post fertilization culture medium would affect the early embryo development and the quality of the produced blastocysts in terms of mRNA expression of several important genes. In a previous study we had shown that media modified with 0.01 mm of G provided the same antioxidant protection as the respective in vivo environments (i.e. the follicular and the oviductal fluid). Bovine cumulus–oocyte complexes (COC) were aspirated from ovaries derived from slaughtered cows and matured in groups of 50 in 500 μL in TCM199 with 10% fetal calf serum and 10 ng mL–1 Epidermal Growth factor at 39°C in an atmosphere of 5% CO2 in air and maximum humidity. Twenty-four hours later matured oocytes were inseminated with frozen/thawed bull semen and co-incubated in the same conditions as maturation. Presumptive zygotes were divided into 4 groups and cultured in groups of 25 in 25 μL of SOF with 5% FCS (Control–, n = 355), supplemented with 0.01 mm of G (n = 344) or 0.1 mm of G (n = 345) or 0.05% DMSO – the G diluent–(Control+, n = 347) at 39°C in an atmosphere of 5% CO2, 5% O2 and maximum humidity. Blastocyst yield was recorded on Days 6, 7, 8 and 9; Day 7 blastocysts from each group were snap frozen and stored at –80°C for mRNA extraction. Quantification of transcripts for aldose reductase mRNA (AKRIBI), prostaglandin G/H synthase-2 (PGHS-2, COX-2), glyceraldehyde 3-phosphate dehydrogenase (GADPH), facilitated glucose/fructose transporter, member 5 (GLUT-5) genes related to metabolism, glutathione peroxidase 1 (GPX1), glucose-6-phosphate dehydrogenase (G6PD) antioxidant enzymes and placenta-specific 8 (PLAC8) related to implantation was carried out by real-time quantitative RT-PCR. Data for embryo development and on transcript abundance were analyzed by chi square and ANOVA, respectively. Cleavage rate tended to be higher in 0.01 mm group than in Control– (77.87% v. 71.41%, P = 0.07). Barring that, no other differences were detected in cleavage rate (Control+: 71.32%; 0.1 mm: 72.75%) or in the overall blastocyst yield on Day 9 (Control–: 25.50%; Control+: 26.71%; 0.1 mm: 25.75%; 0.01 mm: 29.58%). The relative abundance of genes studied varied among groups, but these differences were not significant. We infer that under the current culture conditions, G as an antioxidant has no serious direct effect on early embryo development or on embryo quality at least on the mRNA transcripts studied. Further studies using the same antioxidant in different atmospheric conditions are planed. ED and GSA were sponsored by COST (FAO702) and OECD fellowships, respectively.

scholarly journals The Effects of Aquilaria Malaccensis Leaves Aqueous Extract on The Sperm of Sprague Dawley Rats Towards Early Embryogenesis

2018 ◽  
Vol 17 (1) ◽  
Author(s):  
Faridah Ismail ◽  
Azantee Yazmie Abdul Wahab ◽  
Muhammad Lokman Md Isa ◽  
Redzuan Nul Hakim Abdul Razak ◽  
Raja Arif Shah Raja Ismail
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Treated Group ◽  
Early Embryo ◽  
Male Rat ◽  
Sprague Dawley ◽  
Early Embryo Development ◽  
Aquilaria Malaccensis ◽  
Sprague Dawley Rats

Introduction: Oxidative stress induced by excessive and unopposed levels of reactive oxygen species in male reproductive system results in impaired sperm quality, fertilization capacity and poor embryo development. Our goal is to assess the potential effects of Aquilaria malaccensis (AM) leaves, a plant with strong antioxidant property on early embryo development in vitro and embryo quality following fertilization with cyclophosphamide (CP) exposed rat sperm. Materials and Methods: Forty eight male Sprague Dawley rats were allocated into eight groups of six rats (n = 6): control, CP only (200 mg/kg), AM only (100 mg/kg, 300 mg/kg and 500 mg/kg) and CP + AM (100 mg/kg, 300 mg/kg and 500 mg/kg). Animals were sacrificed after 63 days of treatment and sperm from caudal epididymis were taken for in vitro fertilization (IVF) with oocytes from untreated female. Fertilization, embryo division and embryo morphology were examined after 24 hours post fertilization and compared between groups. Statistical evaluations were performed using Chi-Square test and Fisher’s exact test and pvalue < 0.05 was considered significant. Results: Co-administration of AM leaves extract at dose 100 mg/kg/day to CP-treated group has significantly increased (p < 0.05) the fertilization rate, early cleavage rate and embryo quality compared to CP only treated group. Conclusion: Overall, the present results indicate the potential of AM leaves extract supplementation to improve the fertility and early embryo development in male rat exposed to CP by inhibiting the oxidative processes and scavenging free radicals.

scholarly journals The Effects of Aquilaria malaccensis Leaves Aqueous Extract on Sperm of Sprague Dawley Rats towards Early Embryogenesis

2020 ◽  
Vol 18 (2) ◽  
Author(s):  
Faridah Ismail ◽  
Azantee Yazmie Abdul Wahab ◽  
Muhammad Lokman Md Isa ◽  
Hussin Muhammad ◽  
Raja Arif Shah Raja Ismail ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Early Embryo ◽  
Male Rat ◽  
Embryo Morphology ◽  
Sprague Dawley ◽  
Early Embryo Development ◽  
Aquilaria Malaccensis ◽  
Sprague Dawley Rats

Introduction: Oxidative stress induced by excessive and unopposed levels of reactive oxygen species in male reproductive system results in impaired sperm quality, fertilization capacity and poor embryo development. Our goal is to assess the potential effects of Aquilaria malaccensis (AM) leaves, a plant with strong antioxidant property on early embryo development in vitro and embryo quality following fertilization with cyclophosphamide (CP) exposed rat sperm. Materials and Methods: Twenty four male Sprague Dawley rats were allocated into eight groups of three rats (n = 3): control, CP only (200 mg/kg), AM only (100 mg/kg, 300 mg/kg and 500 mg/kg) and CP + AM (100 mg/kg, 300 mg/kg and 500 mg/kg). Animals were sacrificed after 63 days of treatment and sperm from caudal epididymis were taken for in vitro fertilization (IVF) with oocytes from untreated female. Fertilization, embryo division and embryo morphology were examined at 8 and 48 hours post insemination and compared between groups. Statistical evaluations were performed using Chi-Square test and Fisher’s exact test and p-value <0.05 was considered significant. Results: Administration of AM leave extract at 100 mg/kg/day to normal rats and CP-exposed rats has significantly increased (p><0.05) the fertilization rate, early cleavage rate and embryo quality when compared to CP only treated group. However, other groups showed no significant differences. Conclusion: Overall, the present results indicate the potential of AM leave extract supplementation to improve the fertility and early embryo development in male rat exposed to CP by inhibiting the oxidative processes and scavenging free radicals.>

The Effect of CRH and Its Inhibitor, Antalarmin, on in Vitro Growth of Preantral Mouse Follicles, Early Embryo Development, and Steroidogenesis

Endocrinology ◽  
2013 ◽  
Vol 154 (1) ◽  
pp. 222-231 ◽  
Author(s):  
V. Dinopoulou ◽  
G. A. Partsinevelos ◽  
D. Mavrogianni ◽  
E. Anagnostou ◽  
P. Drakakis ◽  
...  
Keyword(s):  
Embryo Development ◽  
Early Embryo ◽  
In Vitro Growth ◽  
Early Embryo Development

The effects of brain-derived neurotrophic factor and metformin on in vitro developmental competence of bovine oocytes

Zygote ◽  
2009 ◽  
Vol 17 (3) ◽  
pp. 187-193 ◽  
Author(s):  
So Gun Hong ◽  
Goo Jang ◽  
Hyun Ju Oh ◽  
Ok Jae Koo ◽  
Jung Eun Park ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Embryo Development ◽  
Neurotrophic Factor ◽  
Brain Derived Neurotrophic Factor ◽  
Early Embryo ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Significant Difference

SummaryBrain-derived neurotrophic factor (BDNF) signalling via tyrosine kinase B receptors may play an important role in ovarian development and function. It has been reported that metformin elevates the activity of Tyrosine kinase receptors and may amplify BDNF signalling. The objective of this study was to investigate the effect of BDNF during in vitro maturation (IVM) and/or in vitro culture (IVC) (Experiment 1), and to evaluate the collaborative effect of BDNF and metformin treatment on the developmental competence of bovine in vitro fertilized (IVF) embryos (Experiment 2). In Experiment 1, BDNF, which was added to our previously established IVM systems, significantly increased the proportions of MII oocytes at both 10 ng/ml (86.7%) and 100 ng/ml (85.4%) compared with the control (64.0%). However, there was no statistically significant difference in blastocyst development between the control or BDNF-supplemented groups. In Experiment 2, in order to investigate the effect of BDNF (10 ng/ml) and/or metformin (10−5 M) per se, TCM-199 without serum and hormones was used as the control IVM medium. The BDNF (48.3%) and BDNF plus metformin (56.5%) significantly enhanced the proportions of MII oocytes compared with the control (34.4%). Although, BDNF or metformin alone had no effect in embryo development, BDNF plus metformin significantly improved early embryo development to the 8–16-cell stage compared with the control (16.5 vs. 5.5%). In conclusion, the combination of BDNF and metformin may have a collaborative effect during the IVM period. These results could further contribute to the establishment of a more efficient bovine in vitro embryo production system.

P–181 Morphine regulates BMP4 growth factor and is involved in in-vitro early embryo development and PGCs formation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
I Muñoa ◽  
M Araolaza-Lasa ◽  
I Urizar-Arenaza ◽  
M Gianzo Citores ◽  
N Subiran Ciudad
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Environmental Factors ◽  
Embryo Development ◽  
Early Embryo ◽  
Epigenetic Changes ◽  
Morphine Treatment ◽  
Early Embryo Development

Abstract Study question To elucidate if morphine can alter embryo development. Summary answer Chronic morphine treatment regulates BMP4 growth factor, in terms of gene expression and H3K27me3 enrichment and promotes in-vitro blastocysts development and PGC formation. What is known already BMP4 is a member of the bone morphogenetic protein family, which acts mainly through SMAD dependent pathway, to play an important role in early embryo development. Indeed, BMP4 enhances pluripotency in mouse embryonic stem cells (mESCs) and, specifically, is involved in blastocysts formation and primordial germ cells (PGCs) generation. Although, external morphine influence has been previously reported on the early embryo development, focus on implantation and uterus function, there is a big concern in understanding how environmental factors can cause stable epigenetic changes, which could be maintained during development and lead to health problems. Study design, size, duration First, OCT4-reported mESCs were chronically treated with morphine during 24h, 10–5mM. After morphine removal, mESCs were collected for RNA-seq and H3K27me3 ChIP-seq study. To elucidate the role of morphine in early embryo development, two cell- embryos stage were chronically treated with morphine for 24h and in-vitro cultured up to the blastocyst stage in the absence of morphine. Furthermore, after morphine treatment mESCs were differentiated to PGCs, to elucidate the role of morphine in PGC differentiation. Participants/materials, setting, methods Transcriptomic analyses and H3K27me3 genome wide distribution were carried out by RNA-Sequencing and Chip-Sequencing respectively. Validations were performed by RNA-RT-qPCR and Chip-RT-qPCR. Main results and the role of chance Dynamic transcriptional analyses identified a total of 932 differentially expressed genes (DEGs) after morphine treatment on mESCs, providing strong evidence of a transcriptional epigenetic effect induced by morphine. High-throughput screening approaches showed up Bmp4 as one of the main morphine targets on mESCs. Morphine caused an up-regulation of Bmp4 gene expression together with a decrease of H3K27me3 enrichment at promoter level. However, no significant differences were observed on gene expression and H3K27me3 enrichment on BMP4 signaling pathway components (such as Smad1, Smad4, Smad5, Smad7, Prdm1 and Prmd14) after morphine treatment. On the other hand, the Bmp4 gene expression was also up-regulated in in-vitro morphine treated blastocyst and in-vitro morphine treated PGCs. These results were consistent with the increase in blastocyst rate and PGC transformation rate observed after morphine chronic treatment. Limitations, reasons for caution To perform the in-vitro analysis. Further studies are needed to describe the whole signaling pathways underlying BMP4 epigenetic regulation after morphine treatment. Wider implications of the findings: Our findings confirmed that mESCs and two-cell embryos are able to memorize morphine exposure and promote both blastocyst development and PGCs formation through potentially BMP4 epigenetic regulation. These results provide insights understanding how environmental factors can cause epigenetic changes during the embryo development, leading to alterations and producing health problems/diseases Trial registration number Not applicable

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