Oxidative stress induced by methomyl exposure reduces the quality of early embryo development in mice

Zygote ◽  
2021 ◽  
pp. 1-8
Author(s):  
Daohong He ◽  
Guobo Han ◽  
Xiaomeng Zhang ◽  
Jingyu Sun ◽  
Yongnan Xu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Embryonic Development ◽  
Embryo Development ◽  
Early Embryo ◽  
Early Embryonic Development ◽  
Control Group ◽  
Blastocyst Formation ◽  
Early Embryo Development ◽  
Early Embryos

Summary Methomyl is a widely used carbamate insecticide and environmental oestrogen that has adverse effects on the reproductive system. However, there have been no reports on the effect of methomyl on early embryos in mammals. In this study, we explored the effect of methomyl exposure on the quality of early embryonic development in mice and the possible mechanisms. During in vitro culture, different concentrations of methomyl (10, 20, 30 and 35 μM) were added to mouse zygote medium. The results showed that methomyl had an adverse effect on early embryonic development. Compared with the control group, the addition of 30 μM methomyl significantly reduced the rate of early embryo blastocyst formation. Methomyl exposure can increase oxidative stress and impair mitochondrial function, which may be the cause of blastocyst formation. In addition, we found that methomyl exposure promoted apoptosis and autophagy in mouse blastocysts. The toxic effect of methomyl on early embryos may be the result of oxidative stress induction. Taken together, our results indicate that methomyl can cause embryonic development defects in mice, thereby reducing the quality of early embryo development.

Calcium-sensing receptor (CASR) is involved in porcine in vitro fertilisation and early embryo development

2018 ◽  
Vol 30 (2) ◽  
pp. 391 ◽  
Author(s):  
C. Liu ◽  
Y. Liu ◽  
K. Larsen ◽  
Y. P. Hou ◽  
H. Callesen
Keyword(s):  
Embryo Development ◽  
Early Embryo ◽  
Extracellular Calcium ◽  
In Vitro Fertilisation ◽  
Control Group ◽  
Blastocyst Formation ◽  
Calcium Sensing Receptor ◽  
Early Embryo Development ◽  
Calcium Sensing

It has been demonstrated that extracellular calcium is necessary in fertilisation and embryo development but the mechanism is still not well understood. The present study mainly focussed on the extracellular calcium effector called the calcium-sensing receptor (CASR) and examined its expression in porcine gametes and embryos and its function during fertilisation and early embryo development. By using reverse transcription polymerase chain reaction, CASR was found to be expressed in porcine oocytes, spermatozoa and embryos at different developmental stages. Functionally, medium supplementation with a CASR agonist or an antagonist during in vitro fertilisation (IVF) and in vitro culture (IVC) was tested. During fertilisation, the presence of a CASR agonist increased sperm penetration rate and decreased polyspermy rate leading to an increased normal fertilisation rate. During embryo development, for the IVF embryos, agonist treatment during IVC significantly increased cleavage rate and blastocyst formation rate compared with the control group. Furthermore, parthenogenetically activated embryos showed similar results with lower cleavage and blastocyst formation rates in the antagonist group than in the other groups. It was concluded that CASR, as the effector of extracellular calcium, modulates porcine fertilisation and early embryo development.

Nutritional Status Impacts Epigenetic Regulation in Early Embryo Development: A Scoping Review

2021 ◽  
Author(s):  
Shuang Cai ◽  
Shuang Quan ◽  
Guangxin Yang ◽  
Meixia Chen ◽  
Qianhong Ye ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Embryo Development ◽  
Epigenetic Regulation ◽  
Scoping Review ◽  
Epigenetic Modification ◽  
Reproductive Technologies ◽  
Early Embryo ◽  
Epigenetic Modifications ◽  
Nutrition Status ◽  
Early Embryo Development

ABSTRACTWith the increasing maternal age and the use of assisted reproductive technology in various countries worldwide, the influence of epigenetic modification on embryonic development is increasingly notable and prominent. Epigenetic modification disorders caused by various nutritional imbalance would cause embryonic development abnormalities and even have an indelible impact on health in adulthood. In this scoping review, we summarize the main epigenetic modifications in mammals and the synergies among different epigenetic modifications, especially DNA methylation, histone acetylation, and histone methylation. We performed an in-depth analysis of the regulation of various epigenetic modifications on mammals from zygote formation to cleavage stage and blastocyst stage, and reviewed the modifications of key sites and their potential molecular mechanisms. In addition, we discuss the effects of nutrition (protein, lipids, and one-carbon metabolism) on epigenetic modification in embryos and emphasize the importance of various nutrients in embryonic development and epigenetics during pregnancy. Failures in epigenetic regulation have been implicated in mammalian and human early embryo loss and disease. With the use of reproductive technologies, it is becoming even more important to establish developmentally competent embryos. Therefore, it is essential to evaluate the extent to which embryos are sensitive to these epigenetic modifications and nutrition status. Understanding the epigenetic regulation of early embryo development will help us make better use of reproductive technologies and nutrition regulation to improve reproductive health in mammals.

64 Culture of bovine oocytes and embryos with metabolic hormones concentrations associated with equine metabolic syndrome

2019 ◽  
Vol 31 (1) ◽  
pp. 157
Author(s):  
D. Bresnahan ◽  
E. Carnevale
Keyword(s):  
Gene Expression ◽  
Metabolic Syndrome ◽  
Embryo Development ◽  
Early Embryo ◽  
Blastocyst Formation ◽  
Early Embryo Development ◽  
Metabolic Hormones ◽  
Equine Metabolic Syndrome ◽  
And Control ◽  
The Impact

Maternal metabolic status could affect fertility and early embryo development due to altered concentrations of metabolic hormones. Equine metabolic syndrome (EMS) is a condition in horses associated with obesity and insulin resistance. Equine metabolic syndrome is accompanied by increased concentrations of insulin and leptin and decreased concentrations of adiponectin, in ovarian follicular fluid (FF) and in systemic circulation (SYST). We sought to determine how altered concentrations of insulin, leptin, and adiponectin (ILA), consistent with those in mares with EMS (EMS) or normal mares (normal), would affect blastocyst formation rates, blastocyst gene expression for metabolism and inner cell mass formation (OCT4, SOX2, COX2, DNMT3a1, HK2, LDH, PDH, and GLUT1), and metabolite uptake from culture media. Because equine oocytes are not available for large-scale study, a bovine model was used in this preliminary study to determine the impact of altered ILA on oocytes and embryos. Bovine ovaries were obtained from an abattoir and embryos produced as previously described using chemically defined media (CDM; Barcelo-Fimbres and Seidel 2007Mol. Reprod. Dev. 74, 1406-1418). Briefly, oocytes were cultured in in vitro maturation medium (IVM), fertilized in FCDM, presumptive zygotes were placed into CDM-1 for ~56h. Cleavage rates were assessed, and embryos were moved to CDM-2 for ~122 additional hours. Treatments consisted of 5 groups: (1) standard oocyte IVM, FCDM and embryo production (EP) system (control), (2) IVM with normal FF ILA and control FCDM and EP, (3) IVM with normal FF ILA and FCDM and EP with normal SYST ILA, (4) IVM with EMS FF ILA and control FCDM and EP, and (5) IVM with EMS FF ILA and FCDM and EP with EMS SYST ILA. Seven days after fertilization, blastocysts were pooled in groups of 5 and placed into 50mL of CDM-2 for 24h. Embryos were removed, and medium was frozen and stored at −80°C to determine metabolite usage via gas chromatography mass spectroscopy. Pooled embryos were washed and placed into RNA lysis solution for relative quantitative PCR. Statistical comparisons were performed using ANOVA with a post-hoc Tukey test. Blastocyst formation rates and gene expression of viability markers were not significantly different among groups. However, aspartate was lower (P=0.02) in spent media from Group 3 (normal FF and SYST ILA) and tended (P=0.09) to be lower in media from Group 5 (EMS FF and SYST ILA) when compared with controls (Group 1). The ILA during early embryo development but not oocyte maturation appeared to be associated with increased uptake of aspartate, a nonessential amino acid, thought to be involved in osmoregulation, cellular signalling, and in mouse embryos, facilitate the metabolism of lactate. In conclusion, the addition of ILA in concentrations observed in normal horses and EMS horses did not affect blastocyst formation rates or markers of embryo viability, although embryo metabolism could have been altered.

Dynamics of paternal contributions to early embryo development in large animals

2020 ◽  
Author(s):  
Bradford W Daigneault
Keyword(s):  
Embryonic Development ◽  
Embryo Development ◽  
Chromatin Structure ◽  
Current Knowledge ◽  
Domestic Animals ◽  
Experimental Models ◽  
Early Embryo ◽  
Early Embryo Development ◽  
Genomic Studies ◽  
Large Animals

Abstract This review focuses on current knowledge of paternal contributions to preimplantation embryonic development with particular emphasis on large animals. Specifically, the included content aims to summarize genomic and epigenomic contributions of paternally expressed genes, their regulation, and chromatin structure that are indispensable for early embryo development. The accumulation of current knowledge will summarize conserved allelic function among species to include functional molecular and genomic studies across large domestic animals in context with reference to founding experimental models.

228 EFFECT OF INSULIN ON BOVINE OOCYTE MATURATION, CUMULUS CELL APOPTOSIS, AND EARLY EMBRYO DEVELOPMENT IN VITRO

2015 ◽  
Vol 27 (1) ◽  
pp. 203
Author(s):  
I. Lindgren ◽  
P. Humblot ◽  
D. Laskowski ◽  
Y. Sjunnesson
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Insulin Treatment ◽  
Cumulus Cells ◽  
Early Embryo ◽  
Blastocyst Formation ◽  
Early Embryo Development ◽  
Maturation In Vitro

Dairy cow fertility has decreased during the last decades, and much evidence indicates that metabolic disorders are an important part of this decline. Insulin is a key factor in the metabolic challenge during the transition period that coincides with the oocyte maturation and may therefore have an impact on the early embryo development. The aim of this study was to test the effect of insulin during oocyte maturation on early embryo development by adding insulin during the oocyte maturation in vitro. In this study, abattoir-derived bovine ovaries were used and cumulus-oocyte complexes (n = 991) were in vitro matured for 22 h according to standard protocols. Insulin was added during maturation in vitro as follows: H (10 µg mL–1 of insulin), L (0.1 µg mL–1 of insulin), or Z (0 µg mL–1 of insulin). After maturation, oocytes were removed and fixed in paraformaldehyde before staining. Click-it TUNEL assay (Invitrogen, Stockholm, Sweden) was used for apoptotic staining and DRAQ5 (BioNordika, Stockholm, Sweden) for nuclear staining (n = 132). Cumulus-oocyte complexes were evaluated using laser scanning confocal microscope (Zeiss LSM 510, Zeiss, Oberkochen, Germany). Five levels of scans were used to assess oocyte maturation (MII stage) and apoptosis. Because of incomplete penetration of the TUNEL stain (3–5 layers of cumulus cells), only the outer 2 layers of the cumulus complex were investigated regarding apoptosis. Apoptotic index was calculated as apoptotic cells/total cells visualised. Remaining oocytes were fertilized and cultured in vitro until Day 8. Day 7 and Day 8 blastocyst formation was assessed as well as blastocyst stage and grade. Effect of insulin treatment on variables was analysed by ANOVA following arc sin √p transformation. Post-ANOVA comparisons between H+L group v. Z were performed by using the contrast option under GLM (Scheffé test). Results are presented as least squares means ± s.e. P-values ≤ 0.05 were considered as statistically significant. Insulin treatment during oocyte maturation in vitro had no significant effect on oocyte nuclear maturation or apoptotic index of the cumulus cells (Z: 0.052 ± 0.025, L: 0.039 ± 0.016, H: 0.077 ± 0.044, P > 0.05). No effect was seen on cleavage rates (Z: 0.85 ± 0.02, L: 0.85 ± 0.02, H: 0.89 ± 0.03, P > 0.05), but insulin treatment significantly decreased Day 7 rates from fertilized oocytes (Z: 0.19 ± 0.02, L: 0.14 ± 0.02, H: 0.12 ± 0.02, P < 0.05). This study also showed a significantly retarded developmental stage and decreased grade of blastocysts in insulin-treated groups taken together when compared with the control group (P < 0.05). In this study, no effect of insulin supplementation during in vitro maturation was seen on bovine oocyte maturation and apoptosis of cumulus cells, but blastocyst formation and development were negatively affected. Further studies are needed for understanding the relationship between the addition of insulin during maturation in vitro and impaired blastocyst formation. Insulin is a common supplement in the first phase of the first in vitro maturation medium for pig oocytes and is believed to have a beneficial effect on this species.Funding was received from Stiftelsen Nils Lagerlöfs Fond H12–0051-NLA.

181 Equine androgenic embryos: ability of the equine sperm to develop in a heterospecific ooplasm

2019 ◽  
Vol 31 (1) ◽  
pp. 215
Author(s):  
M. B. Rodriguez ◽  
A. Gambini ◽  
D. F. Salamone
Keyword(s):  
Embryonic Development ◽  
Embryo Development ◽  
Uv Light ◽  
Early Embryo ◽  
Control Treatment ◽  
Cleavage Rate ◽  
Early Embryo Development ◽  
Sperm Dna ◽  
Haploid Embryos ◽  
Androgenic Embryos

Androgenic haploid embryos were originally produced for the study of certain aspects of early embryo development. The generation of androgenic haploid embryos allows us to better understand the complementary parental contribution to embryonic development, and to examine the effects of haploid development on gene expression. Because mare oocytes for research are scarce, the generation of heterospecific androgenic embryos could be useful to study aspects of the biology of early embryo development, or to identify genes and their variations or mutations that are responsible for reproduction-related problems in mares and stallions, which is of interest for the breeding industry. Therefore, the aim of this work was to study the capability of equine sperm to induce embryonic development after injection into an enucleated oocyte from a different species. Porcine cumulus-oocyte complexes (COC) were obtained from abattoir ovaries and placed in 100-µL drops in vitro maturation (IVM) medium for 42h. Cumulus cells were removed with hyaluronidase and vortexing. Then, mature oocytes were subjected to intracytoplasmic sperm injection (ICSI) with stallion frozen-thawed semen (according to Rodriguez et al. 2015). Immediately after the last injection, the zona pellucida of injected oocytes was removed with protease treatment, the oocytes were treated with cytochalasin B, and the metaphase II enucleated with a 20-µm micropipette. Finally, embryos were placed in culture medium (SOF) in plates with the well-of-the-well (WOW) system. As control treatment, non-enucleated pig oocytes were injected with stallion (CE) and boar (CC) semen. At Day 4, embryos were evaluated for cleavage and number of blastomeres, and stained with Hoechst 33342 to verify the presence of DNA in each blastomere under the UV light. Embryos were stored for future PCR studies to validate the presence of equine DNA. Data were analysed by chi-squared test to compare the cleavage of both controls with the androgenic embryos. From a total of 53 androgenic haploid embryos, the cleavage rate was 62% (33/53). Embryos were cleaved in 2 to 4 cells in 72.7%, 5 to 8 cells in 18.2%, and 9+ cells in 9.1% at Day 4. Presence of DNA in all blastomeres was observed in 60.6% (20/33) of the androgenic haploid embryos, while 21.2% (7/33) of the embryos had 10 to 50% of blastomeres with DNA, and 18.6% (6/33) of the embryos did not have DNA in their blastomeres. The ICSI control embryos cleaved in 45.3% (34/75) and 64.9% (98/151) for groups CC and CE, respectively. Cleavage rates in control CE were significantly higher than those in control CC (P&lt;0.004). No statistical difference was observed in the control groups versus androgenic embryos. This preliminary results showed that a heterospecific ooplasm can be successfully used to allow an equine sperm DNA to decondense and to develop, even in absence of the female counterpart. Using this method, copies of a single sperm DNA can be produced to potentially evaluate individual aspects of early embryo development concerning the male contribution. This is the first report of successful androgenic embryos using a heterospecific oocyte to create copies of a horse sperm DNA.

scholarly journals Effect of Triclosan Exposure on Developmental Competence in Parthenogenetic Porcine Embryo during Preimplantation

2020 ◽  
Vol 21 (16) ◽  
pp. 5790
Author(s):  
Min Ju Kim ◽  
Hyo-Jin Park ◽  
Sanghoon Lee ◽  
Hyo-Gu Kang ◽  
Pil-Soo Jeong ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Embryonic Development ◽  
Mitochondrial Dysfunction ◽  
Formation Rate ◽  
Developmental Competence ◽  
Early Embryonic Development ◽  
Toxic Effects ◽  
Blastocyst Formation ◽  
Parthenogenetic Embryos

Triclosan (TCS) is included in various healthcare products because of its antimicrobial activity; therefore, many humans are exposed to TCS daily. While detrimental effects of TCS exposure have been reported in various species and cell types, the effects of TCS exposure on early embryonic development are largely unknown. The aim of this study was to determine if TCS exerts toxic effects during early embryonic development using porcine parthenogenetic embryos in vitro. Porcine parthenogenetic embryos were cultured in in vitro culture medium with 50 or 100 µM TCS for 6 days. Developmental parameters including cleavage and blastocyst formation rates, developmental kinetics, and the number of blastomeres were assessed. To determine the toxic effects of TCS, apoptosis, oxidative stress, and mitochondrial dysfunction were assessed. TCS exposure resulted in a significant decrease in 2-cell rate and blastocyst formation rate, as well as number of blastomeres, but not in the cleavage rate. TCS also increased the number of apoptotic blastomeres and the production of reactive oxygen species. Finally, TCS treatment resulted in a diffuse distribution of mitochondria and decreased the mitochondrial membrane potential. Our results showed that TCS exposure impaired porcine early embryonic development by inducing DNA damage, oxidative stress, and mitochondrial dysfunction.

Quantitative analysis of sperm mRNA in the pig: relationship with early embryo development and capacitation

2013 ◽  
Vol 25 (5) ◽  
pp. 807 ◽  
Author(s):  
Jae Yeon Hwang ◽  
Brendan P. Mulligan ◽  
Hyung-Min Kim ◽  
Byoung-Chul Yang ◽  
Chang-Kyu Lee
Keyword(s):  
Embryo Development ◽  
Early Embryo ◽  
Mrna Abundance ◽  
Mrna Levels ◽  
Cleavage Rate ◽  
Early Embryo Development ◽  
Specific Mrnas ◽  
Porcine Spermatozoa ◽  
Mammalian Spermatozoa

Although it is well known that mRNA is present in mammalian spermatozoa, the relevance of mRNA to capacitation and early embryo development in the pig remains unclear. In the present study, we investigated differences in the abundance of selected mRNAs coding for MYC, CYP19, ADAM2, PRM1 and PRM2 in purified porcine spermatozoa depending on embryo cleavage rate and capacitation (n = 20 semen samples). Semen samples were used in IVF procedures, with subsequent embryo development classified into one of two groups based on cleavage rate (i.e. high (>75%) and low (<75%) cleavage groups) and mRNA abundance in purified spermatozoa compared between these two groups. In addition, mRNA abundance was compared between capacitated and non-capacitated spermatozoa. Comparison of mRNA levels between porcine spermatozoa revealed that the abundance of MYC, CYP19, ADAM2, PRM1 and PRM2 mRNA was significantly greater in the high cleavage group (n = 10 high cleavage group semen samples) than in the low cleavage group (n = 10; P < 0.05). Significant downregulation of MYC mRNA was observed in capacitated spermatozoa (n = 12; P < 0.05). The results of the present study suggest that the amount of specific mRNAs could be used for estimating the quality of spermatozoa in the pig.

Superovulation alters embryonic poly(A)-binding protein (Epab) and poly(A)-binding protein, cytoplasmic 1 (Pabpc1) gene expression in mouse oocytes and early embryos

2016 ◽  
Vol 28 (3) ◽  
pp. 375 ◽  
Author(s):  
Saffet Ozturk ◽  
Aylin Yaba-Ucar ◽  
Berna Sozen ◽  
Derya Mutlu ◽  
Necdet Demir
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Binding Protein ◽  
Early Embryo ◽  
High Dose ◽  
Mrna Levels ◽  
Early Embryo Development ◽  
Female Mice ◽  
Early Embryos

Embryonic poly(A)-binding protein (EPAB) and poly(A)-binding protein, cytoplasmic 1 (PABPC1) play critical roles in translational regulation of stored maternal mRNAs required for proper oocyte maturation and early embryo development in mammals. Superovulation is a commonly used technique to obtain a great number of oocytes in the same developmental stages in assisted reproductive technology (ART) and in clinical or experimental animal studies. Previous studies have convincingly indicated that superovulation alone can cause impaired oocyte maturation, delayed embryo development, decreased implantation rate and increased postimplantation loss. Although how superovulation results in these disturbances has not been clearly addressed yet, putative changes in genes related to oocyte and early embryo development seem to be potential risk factors. Thus, the aim of the present study was to determine the effect of superovulation on Epab and Pabpc1 gene expression. To this end, low- (5 IU) and high-dose (10 IU) pregnant mare’s serum gonadotropin (PMSG) and human chorionic gonadotrophin (hCG) were administered to female mice to induce superovulation, with naturally cycling female mice serving as controls. Epab and Pabpc1 gene expression in germinal vesicle (GV) stage oocytes, MII oocytes and 1- and 2-cell embryos collected from each group were quantified using quantitative reverse transcription–polymerase chain reaction. Superovulation with low or high doses of gonadotropins significantly altered Epab and Pabpc1 mRNA levels in GV oocytes, MII oocytes and 1- and 2-cell embryos compared with their respective controls (P < 0.05). These changes most likely lead to variations in expression of EPAB- and PABPC1-regulated genes, which may adversely influence the quality of oocytes and early embryos retrieved using superovulation.

Sonic hedgehog promotes porcine oocyte maturation and early embryo development

2009 ◽  
Vol 21 (6) ◽  
pp. 805 ◽  
Author(s):  
Ngoc Tan Nguyen ◽  
David Pei-Cheng Lin ◽  
Shih-Ying Yen ◽  
Jung-Kai Tseng ◽  
Jui-Fen Chuang ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sonic Hedgehog ◽  
Oocyte Maturation ◽  
Calcium Release ◽  
Cyclin B1 ◽  
Early Embryo ◽  
Control Group ◽  
Early Embryo Development ◽  
Porcine Oocyte

In the present study, we investigated the effects of the Sonic hedgehog (Shh) protein on porcine oocyte maturation and early embryo development. Immunohistochemistry showed activation of Shh signalling in cumulus–oocyte complexes (COCs), as reflected by Patched (Ptc), Smoothened (Smo) and Gli1 expression in oocytes, cumulus cells and granulosa cells, particularly those of small follicles (<2 mm in diameter). Western blot analysis showed Smo expression in COCs and in denuded oocytes derived from small and medium (3–7 mm)-sized follicles. Small follicles contained the highest concentration of Shh in follicular fluid compared with medium-sized and large (>7 mm in diameter) follicles. Supplementation with Shh (0.5 or 1 μg mL–1) enhanced oocyte maturation compared with the control group (92.4% and 90.4% v. 81.9%, respectively; P < 0.05). This effect was reversed by the simultaneous addition of cyclopamine (1–2 μm), an Shh inhibitor. Similar to intact COCs, denuded COCs showed enhanced maturation following Shh supplementation. Furthermore, cyclin B1 content, extracellular signal-regulated kinase 1/2 phosphorylation, intracellular calcium release, blastocyst rate and total cell numbers were greater (P < 0.05) in oocytes matured in the presence of 0.5 and 1 μg mL–1 Shh compared with control oocytes. The findings of the present study provide the first evidence that the Shh signalling pathway is active, or at least partially activated, in the porcine ovary and is likely to promote oocyte cytoplasmic and nuclear maturation, as well as subsequent in vitro development, although the underlying mechanisms remain to be elucidated.

Sign in / Sign up

Export Citation Format

Share Document