Specific imprinted genes demethylation in association with oocyte donor’s age and culture conditions in bovine embryos assessed at day 7 and 12 post insemination

Lafontaine, Simon; Labrecque, Rémi; Palomino, J. M.; Blondin, Patrick; Sirard, Marc‐André

doi:10.1016/j.theriogenology.2020.09.027

Cited by 9 publications

(13 citation statements)

References 53 publications

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“…At least 20 clones for each sample were picked and sequenced. Validation of the imprinted status of each DMR was performed as previously described by Lafontaine et al (2020), by assessing methylation of sperm DNA (expected methylation >90% or <10%) and fibroblast cell DNA (40-60% expected methylation).…”

Section: Gene Specific Bisulfite Sequencingmentioning

confidence: 99%

Dysregulated Gene Expression of Imprinted and X-Linked Genes: A Link to Poor Development of Bovine Haploid Androgenetic Embryos

Águila

Suzuki

Hill

et al. 2021

Front. Cell Dev. Biol.

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Mammalian uniparental embryos are efficient models for genome imprinting research and allow studies on the contribution of the paternal and maternal genomes to early embryonic development. In this study, we analyzed different methods for production of bovine haploid androgenetic embryos (hAE) to elucidate the causes behind their poor developmental potential. Results indicate that hAE can be efficiently generated by using intracytoplasmic sperm injection and oocyte enucleation at telophase II. Although androgenetic haploidy does not disturb early development up to around the 8-cell stage, androgenetic development is disturbed after the time of zygote genome activation and hAE that reach the morula stage are less capable to reach the blastocyst stage of development. Karyotypic comparisons to parthenogenetic- and ICSI-derived embryos excluded chromosomal segregation errors as causes of the developmental constraints of hAE. However, analysis of gene expression indicated abnormal levels of transcripts for key long non-coding RNAs involved in X chromosome inactivation and genomic imprinting of the KCNQ1 locus, suggesting an association with X chromosome and some imprinted loci. Moreover, transcript levels of methyltransferase 3B were significantly downregulated, suggesting potential anomalies in hAE establishing de novo methylation. Finally, the methylation status of imprinted control regions for XIST and KCNQ1OT1 genes remained hypomethylated in hAE at the morula and blastocyst stages, confirming their origin from spermatozoa. Thus, our results exclude micromanipulation and chromosomal abnormalities as major factors disturbing the normal development of bovine haploid androgenotes. In addition, although the cause of the arrest remains unclear, we have shown that the inefficient development of haploid androgenetic bovine embryos to develop to the blastocyst stage is associated with abnormal expression of key factors involved in X chromosome activity and genomic imprinting.

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Section: Gene Specific Bisulfite Sequencingmentioning

confidence: 99%

Dysregulated Gene Expression of Imprinted and X-Linked Genes: A Link to Poor Development of Bovine Haploid Androgenetic Embryos

Águila

Suzuki

Hill

et al. 2021

Front. Cell Dev. Biol.

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“…Moreover, molecular evidence highlighted transcriptomic (Morin-Doré et al, 2017) and epigenetic (Morin-Doré et al, 2020) differences between embryos derived from mature and immature oocyte donors. In a previous study, we also observed that the donor's age coupled with ART affected maternally imprinted genes in embryonic and trophoblastic cells of Days 7 and 12 embryos (Lafontaine et al, 2020).…”

Section: Introductionmentioning

confidence: 69%

IGF2R, KCNQ1, PLAGL1, and SNRPN DNA methylation is completed in bovine by the early antral follicle stage

Lafontaine

Sirard

2022

Molecular Reproduction Devel

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Imprinted genes are inherited with different DNA methylation patterns depending on the maternal or paternal origin of the allele. In cattle (Bos taurus), abnormal methylation of these genes is linked to the large offspring syndrome, a neonatal overgrowth phenotype analogous to the human Beckwith–Wiedemann syndrome. We hypothesized that in bovine oocytes, some of the methylation patterns on maternally imprinted genes are acquired in the last phase of folliculogenesis. The pyrosequencing analysis of IGF2R, KCNQ1, PLAGL1, and SNRPN imprinted genes showed no clear progression of methylation in oocytes from follicles 1–2 mm (late pre antral/early antral) and up. Instead, these oocytes displayed complete methylation at the imprinted differentially methylated regions (>80%). Other mechanisms related to imprint maintenance should be investigated to explain the hypomethylation at IGF2R, KCNQ1, PLAGL1, and SNRPN maternally imprinted sites observed in some bovine embryos.

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“…Genes expression is often regulated by DNA cytosine methylation, catalyzed by DNA methyltransferases, and it is often altered during in vitro culture (Lafontaine et al, 2020). Owing to the dysregulated transcript expression observed in hAE, specifically in genes from the KCNQ1 locus and X chromosome, we analyzed the expression of enzymes related to maintenance (DNMT1) and de novo (DNMT3B) DNA methylation as well as demethylation (TET1) in haploid (parthenogenetic and androgenetic) and biparental (ICSI) morula stage embryos.…”

Section: Resultsmentioning

confidence: 99%

“…16 clones for each sample were picked and sequenced. Validation of the imprinted status of each DMR was performed as previously described by Lafontaine et al (2020), by assessing methylation of sperm DNA (expected methylation > 90% or <10%) and fibroblast cell DNA (40-60% expected methylation).…”

Section: Methodsmentioning

confidence: 99%

Dysregulated gene expression of imprinted and X-linked genes: a link to poor development of bovine haploid androgenetic embryos

Águila

Therrien

Suzuki

et al. 2020

Preprint

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Mammalian uniparental embryos are efficient models for genome imprinting research and allow studies on the contribution of the paternal and maternal genome to early embryonic development. In this study, we analyzed different methodologies for production of bovine haploid androgenetic embryos (hAE) to elucidate the causes behind their poor developmental potential. The results showed that hAE can be efficiently generated by using intracytoplasmic sperm injection and oocyte enucleation at telophase II. Although haploidy does not disturb early development up to around the 3rd mitotic division, androgenetic development is disturbed after the time of zygote genome activation those that reach the morula stage are less capable to become a blastocyst. Analysis of gene expression indicated abnormal levels of methyltransferase 3B and key long non-coding RNAs involved in X-chromosome inactivation and genomic imprinting of the KCNQ1 locus, which is associated to the methylation status of imprinted control regions of XIST and KCNQ1OT1. Thus, our results seem to exclude micromanipulation consequences and chromosomal abnormalities as major factors in developmental restriction, suggesting that their early developmental constraint is regulated at an epigenetic level.

show abstract

Specific imprinted genes demethylation in association with oocyte donor’s age and culture conditions in bovine embryos assessed at day 7 and 12 post insemination

Cited by 9 publications

References 53 publications

Dysregulated Gene Expression of Imprinted and X-Linked Genes: A Link to Poor Development of Bovine Haploid Androgenetic Embryos

Dysregulated Gene Expression of Imprinted and X-Linked Genes: A Link to Poor Development of Bovine Haploid Androgenetic Embryos

IGF2R, KCNQ1, PLAGL1, and SNRPN DNA methylation is completed in bovine by the early antral follicle stage

Dysregulated gene expression of imprinted and X-linked genes: a link to poor development of bovine haploid androgenetic embryos

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