Decrease in DNA methylase activity during preimplantation development in the mouse

Development ◽  
1991 ◽  
Vol 112 (1) ◽  
pp. 189-192 ◽  
Author(s):  
M. Monk ◽  
R.L. Adams ◽  
A. Rinaldi
Keyword(s):  
Dna Methylation ◽  
Dna Methyltransferase ◽  
Large Scale ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Mouse Development ◽  
Dna Methylase ◽  
Early Mouse ◽  
Absolute Decrease ◽  
Methylase Activity

During early mouse development, there are large-scale changes in DNA methylation. These changes may be due to the availability or stability of the enzyme, DNA methyltransferase (methylase), which is responsible for maintenance of DNA methylation. A microassay for methylase activity in preimplantation embryos shows that the level of maternally inherited enzyme is extremely high in the egg and that this activity is stable for the first three cleavage divisions. However, from the 8-cell to the blastocyst stage, there is a marked and absolute decrease in enzyme activity.

Targets and dynamics of promoter DNA methylation during early mouse development

2010 ◽  
Vol 42 (12) ◽  
pp. 1093-1100 ◽  
Author(s):  
Julie Borgel ◽  
Sylvain Guibert ◽  
Yufeng Li ◽  
Hatsune Chiba ◽  
Dirk Schübeler ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Mouse Development ◽  
Promoter Dna Methylation ◽  
Promoter Dna ◽  
Early Mouse

scholarly journals Dnmt1 binds and represses genomic retroelements via DNA methylation in mouse early embryos

2020 ◽  
Vol 48 (15) ◽  
pp. 8431-8444 ◽  
Author(s):  
Byungkuk Min ◽  
Jung Sun Park ◽  
Young Sun Jeong ◽  
Kyuheum Jeon ◽  
Yong-Kook Kang
Keyword(s):  
Dna Methylation ◽  
Cpg Islands ◽  
Endogenous Retrovirus ◽  
Dna Demethylation ◽  
Mouse Development ◽  
Genome Wide ◽  
Early Embryos ◽  
Dna Adenine Methylase ◽  
Early Mouse ◽  
Early Developmental

Abstract Genome-wide passive DNA demethylation in cleavage-stage mouse embryos is related to the cytoplasmic localization of the maintenance methyltransferase DNMT1. However, recent studies provided evidences of the nuclear localization of DNMT1 and its contribution to the maintenance of methylation levels of imprinted regions and other genomic loci in early embryos. Using the DNA adenine methylase identification method, we identified Dnmt1-binding regions in four- and eight-cell embryos. The unbiased distribution of Dnmt1 peaks in the genic regions (promoters and CpG islands) as well as the absence of a correlation between the Dnmt1 peaks and the expression levels of the peak-associated genes refutes the active participation of Dnmt1 in the transcriptional regulation of genes in the early developmental period. Instead, Dnmt1 was found to associate with genomic retroelements in a greatly biased fashion, particularly with the LINE1 (long interspersed nuclear elements) and ERVK (endogenous retrovirus type K) sequences. Transcriptomic analysis revealed that the transcripts of the Dnmt1-enriched retroelements were overrepresented in Dnmt1 knockdown embryos. Finally, methyl-CpG-binding domain sequencing proved that the Dnmt1-enriched retroelements, which were densely methylated in wild-type embryos, became demethylated in the Dnmt1-depleted embryos. Our results indicate that Dnmt1 is involved in the repression of retroelements through DNA methylation in early mouse development.

scholarly journals Repeats as global DNA methylation marker in bovine preimplantation embryos

2017 ◽  
Vol 62 (No. 2) ◽  
pp. 43-50 ◽  
Author(s):  
W. Li ◽  
A. Van Soom ◽  
L. Peelman
Keyword(s):  
Dna Methylation ◽  
Mutation Rate ◽  
Methylation Level ◽  
Methylation Status ◽  
Repetitive Sequences ◽  
Cell Types ◽  
Blastocyst Stage ◽  
Repeat Sequence ◽  
Preimplantation Embryos ◽  
Global Dna Methylation

DNA methylation undergoes dynamic changes and is a crucial part of the epigenetic regulation during mammalian early development. To determine the DNA methylation levels in bovine embryos, we applied a bisulfite sequencing based method aimed at repetitive sequences including three retrotransposons (L1_BT, BovB, and ERV1-1-I_BT) and Satellite I. A more accurate estimate of the global DNA methylation level compared to previous methods using only one repeat sequence, like Alu, could be made by calculation of the weighted arithmetic mean of multiple repetitive sequences, considering the copy number of each repetitive sequence. Satellite I and L1_BT showed significant methylation reduction at the blastocyst stage, while BovB and ERV1-1-I_BT showed no difference. The mean methylation level of the repetitive sequences during preimplantation development was the lowest at the blastocyst stage. No methylation difference was found between embryos cultured in 5% and 20% O<sub>2</sub>. Because mutations of CpGs negatively influence the calculation accuracy, we checked the mutation rate of the sequenced CpG sites. Satellite I and L1_BT showed a relatively low mutation rate (1.92 and 3.72% respectively) while that of ERV1-1-I_BT and BovB was higher (11.95 and 24% respectively). Therefore we suggest using a combination of repeats with low mutation rate, taking into account the proportion of each sequence, as a relatively quick marker for the global DNA methylation status of preimplantation stages and possibly also for other cell types.

Expression of SRG3, a chromatin-remodelling factor, in the mouse oocyte and early preimplantation embryos

Zygote ◽  
2007 ◽  
Vol 15 (2) ◽  
pp. 129-138 ◽  
Author(s):  
F. Sun ◽  
F. Tang ◽  
A Y. Yan ◽  
H. Y. Fang ◽  
H. Z. Sheng
Keyword(s):  
Essential Role ◽  
Structure And Function ◽  
Chromatin Remodelling ◽  
Preimplantation Embryos ◽  
Protein Synthesis Inhibitor ◽  
Mouse Development ◽  
Synthesis Inhibitor ◽  
Female Pronucleus ◽  
Early Mouse ◽  
And Function

SummarySRG3 (Smarcc1) is a core subunit of the SWI/SNF complex. In the absence of SRG3, embryonic development ceases during peri-implantation stages, indicating that SRG3, as well as the chromatin-remodelling process, plays an essential role in early mouse development. To gain a better understanding of chromatin remodelling during the early stages of development, we examined SRG3 expression during oogenesis and preimplantation stages using immunofluorescence and western blot assays. SRG3 was detected in nuclei of oocytes during growth and maturation. Following fertilization, SRG3 was detected in pronuclei shortly after their formation. Nuclear concentrations of SRG3 increased in a time-dependent fashion and were found to be greater in the male pronucleus than in the female pronucleus. The increase in nuclear SRG3 was partially inhibited by a protein synthesis inhibitor, but not by a transcriptional inhibitor. Expression of SRG3 is accompanied by expression of Brg1 and Ini1, two other core subunits of the SWI/SNF complex. The expression of these three remodelling factors parallels that of SP1 and TBP, both spatially and temporally, in the mouse embryo, suggesting a role for remodelling factors in chromatin structure and function during early development.

scholarly journals G9a regulates temporal preimplantation developmental program and lineage segregation in blastocyst

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Jan J Zylicz ◽  
Maud Borensztein ◽  
Frederick CK Wong ◽  
Yun Huang ◽  
Caroline Lee ◽  
...  
Keyword(s):  
Cell Fate ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Vital Role ◽  
Mouse Development ◽  
Cell Stage ◽  
Developmental Progression ◽  
Early Mouse ◽  
The Impact

Early mouse development is regulated and accompanied by dynamic changes in chromatin modifications, including G9a-mediated histone H3 lysine 9 dimethylation (H3K9me2). Previously, we provided insights into its role in post-implantation development (Zylicz et al., 2015). Here we explore the impact of depleting the maternally inherited G9a in oocytes on development shortly after fertilisation. We show that G9a accumulates typically at 4 to 8 cell stage to promote timely repression of a subset of 4 cell stage-specific genes. Loss of maternal inheritance of G9a disrupts the gene regulatory network resulting in developmental delay and destabilisation of inner cell mass lineages by the late blastocyst stage. Our results indicate a vital role of this maternally inherited epigenetic regulator in creating conducive conditions for developmental progression and on cell fate choices.

scholarly journals Developmental and molecular correlates of bovine preimplantation embryos

Reproduction ◽  
2006 ◽  
Vol 131 (5) ◽  
pp. 895-904 ◽  
Author(s):  
Hakan Sagirkaya ◽  
Muge Misirlioglu ◽  
Abdullah Kaya ◽  
Neal L First ◽  
John J Parrish ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methyltransferase ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Preimplantation Embryos ◽  
Developmental Potential

Expression of embryonic genes is altered in different culture conditions, which influence developmental potential both during preimplantation and fetal development. The objective of this study was to define the effects of culture conditions on: bovine embryonic development to blastocyst stage, blastocyst cell number, apoptosis and expression patterns of a panel of developmentally important genes. Bovine embryos were culturedin vitroin three culture media containing amino acids, namely potassium simplex optimization medium (KSOMaa), Charles Rosenkrans 1 (CR1aa) and synthetic oviductal fluid (SOFaa). Apoptosis in blastocysts was determined by TUNEL assay and expression profiles of developmentally important genes were assayed by real-time PCR.In vivo-produced bovine blastocysts were used as controls for experiments determining gene expression patterns. While the cleavage rates did not differ, embryos cultured in SOFaa had higher rates of development to blastocyst stage (P< 0.05). Mean cell numbers and percentages of apoptotic cells per blastocyst did not differ among the groups. Expression of the heat shock protein 70 (Hsp70) gene was significantly up-regulated in both CR1aa and KSOMaa when compared with SOFaa (P< 0.001). DNA methyltransferase 3a (Dnmt3a) expression was higher in embryos cultured in CR1aa than in those cultured in SOFaa (P< 0.001). Expression of interferon tau (IF-τ) and insulin-like growth factor II receptor (Igf-2r) genes was significantly up-regulated in KSOMaa when compared with CR1aa (P< 0.001). Gene expression did not differ betweenin vivo-derived blastocysts and theirin vitro-derived counterparts. In conclusion, SOFaa supports higher development to blastocyst stage than KSOMaa and CR1aa, and the culture conditions influence gene expression.

scholarly journals H3K27me3 at Pericentromeric Heterochromatin is a Defining Feature of the Early Mouse Blastocyst

2022 ◽  
Author(s):  
Mélanie Pailles ◽  
Mélanie Hirlemann ◽  
Vincent Brochard ◽  
Martine Chebrout ◽  
Jean-François Oudin ◽  
...  
Keyword(s):  
Blastocyst Stage ◽  
Pericentromeric Heterochromatin ◽  
Embryonic Cells ◽  
Mouse Development ◽  
Mouse Blastocyst ◽  
Major Satellite ◽  
Post Translational Modifications ◽  
Early Mouse ◽  
Diffuse Distribution

Abstract Early mouse development is characterized by structural and epigenetic changes at the chromatin level while cells progress towards differentiation. At blastocyst stage, the segregation of the three primordial lineages is accompanied by establishment of differential patterns of DNA methylation and post-translational modifications of histones, such as H3K27me3. In this study, we have analysed the dynamics of H3K27me3 at pericentromeric heterochromatin (PCH) during development of the mouse blastocyst, in comparison with cultured embryonic cells. We show that this histone modification is first enriched at PCH in the whole embryo and evolves into a diffuse distribution in epiblast during its specification and maturation. Concomitantly, the level of transcription from major satellite decreases. Stem cells derived from blastocyst (naïve ESCs and TSCs) do not fully maintain the H3K27me3 enrichment at PCH. Moreover, the dynamic of H3K27me3 at PCH during in vitro conversion from naïve to primed pluripotent state and during ESCs derivation suggests that the mechanisms underlying the control of this histone mark at PCH are different in embryo and in vitro. We also conclude that the non-canonical presence of H3K27me3 at PCH is a defining feature of embryonic cells in the young blastocyst before epiblast segregation.

36 EFFECT OF DNA METHYLTRANSFERASE INHIBITOR, RG108, ON IN VITRO DEVELOPMENT AND ntES ESTABLISHMENT RATE IN CLONED MOUSE EMBRYOS

2012 ◽  
Vol 24 (1) ◽  
pp. 130 ◽  
Author(s):  
C. Li ◽  
Y. Terashita ◽  
M. Tokoro ◽  
S. Wakayama ◽  
T. Wakayama
Keyword(s):  
Dna Methylation ◽  
Histone Acetylation ◽  
Cell Lines ◽  
Dna Methyltransferase ◽  
Histone Deacetylase Inhibitors ◽  
Trichostatin A ◽  
Blastocyst Stage ◽  
Control Group ◽  
Cloning Efficiency ◽  
Dna Methyltransferase Inhibitor

Somatic cell nuclear transfer technique increased expectations among many for its potential to advance the regenerative therapy field. Cloned embryos, however, exhibit several epigenetic abnormalities, such as low histone acetylation or high DNA methylation levels compared with normal fertilized embryos. Therefore, increasing histone acetylation or reducing DNA methylation levels in cloned embryos using chemical treatments may improve cloning efficiency. We recently succeeded in improving the success rate of mouse cloning by using class IIb histone deacetylase inhibitors, such as trichostatin A (TSA), scriptaid and suberoylanilide hydroxamic acid. It has also been reported that 5-aza-2′-deoxycytidine, a DNA methyltransferase inhibitor that is a chemical analogue of cytidine, inhibits the potential of embryos to develop into blastocysts and later to fetuses. In the present study, another DNA methyltransferase inhibitor RG108, which is thought to strongly interact with the DNMT1 active site to inhibit DNMT1 activity, was used to examine whether it could improve cloning efficiency. To determine the effects of RG108, cloned embryos were treated with 100 to 500 μM RG108. When cloned embryos were treated at the 1-cell stage (from artificial activation to 10 h, n = 219), the cloning efficiency was similar to the control group (8.2 vs 10.8%). On the other hand, when 500 μM RG108 was added to the culture medium from the 2-cell to morula/blastocyst stage (n = 113), although the developmental rate to blastocyst stage did not change significantly (79.6% vs 72.3%), higher Oct3/4 expression and more ICM cells were observed compared with non-treated, control cloned embryos. Moreover, we tried to establish ES cell lines from those cloned embryos and 11 ntES lines were generated from 21 blastocysts, which was higher than that of control (6 ntES cell lines from 20 blastocysts). All ntES lines showed AP staining positively. This finding showed that the quality of cloned mouse blastocysts increased when treated with a DNA methyltransferase inhibitor, suggesting a possible means for improving cloning efficiency in the future.

Exposure to mono-n-butyl phthalate disrupts the development of preimplantation embryos

2013 ◽  
Vol 25 (8) ◽  
pp. 1174 ◽  
Author(s):  
Da-Peng Chu ◽  
Shi Tian ◽  
Da-Guang Sun ◽  
Chan-Juan Hao ◽  
Hong-Fei Xia ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Dna Methylation ◽  
Dibutyl Phthalate ◽  
Preimplantation Embryo ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Oxygen Species ◽  
Preimplantation Embryo Development ◽  
Immunofluorescent Analysis ◽  
Butyl Phthalate

Dibutyl phthalate (DBP), a widely used phthalate, is known to cause many serious diseases, especially in the reproductive system. However, little is known about the effects of its metabolite, mono-n-butyl phthalate (MBP), on preimplantation embryo development. In the present study, we found that treatment of embryos with 10–3 M MBP impaired developmental competency, whereas exposure to 10–4 M MBP delayed the progression of preimplantation embryos to the blastocyst stage. Furthermore, reactive oxygen species (ROS) levels in embryos were significantly increased following treatment with 10–3 M MBP. In addition, 10–3 M MBP increased apoptosis via the release of cytochrome c, whereas immunofluorescent analysis revealed that exposure of preimplantation embryos to MBP concentration-dependently (10–5, 10–4 and 10–3 M) decreased DNA methylation. Together, the results indicate a possible relationship between MBP exposure and developmental failure in preimplantation embryos.

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