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DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development

BACKGROUND: In mammals, maternal differentially methylated regions (DMRs) acquire DNA methylation during the postnatal growth stage of oogenesis, with paternal DMRs acquiring DNA methylation in the perinatal prospermatagonia. Following fusion of the male and female gametes, it is widely accepted tha...

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Autores principales: O’Doherty, Alan M, Magee, David A, O’Shea, Lynee C, Forde, Niamh, Beltman, Marijke E, Mamo, Solomon, Fair, Trudee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4363183/
https://www.ncbi.nlm.nih.gov/pubmed/25881176
http://dx.doi.org/10.1186/s12861-015-0060-2
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author O’Doherty, Alan M
Magee, David A
O’Shea, Lynee C
Forde, Niamh
Beltman, Marijke E
Mamo, Solomon
Fair, Trudee
author_facet O’Doherty, Alan M
Magee, David A
O’Shea, Lynee C
Forde, Niamh
Beltman, Marijke E
Mamo, Solomon
Fair, Trudee
author_sort O’Doherty, Alan M
collection PubMed
description BACKGROUND: In mammals, maternal differentially methylated regions (DMRs) acquire DNA methylation during the postnatal growth stage of oogenesis, with paternal DMRs acquiring DNA methylation in the perinatal prospermatagonia. Following fusion of the male and female gametes, it is widely accepted that murine DNA methylation marks at the DMRs of imprinted genes are stable through embryogenesis and early development, until they are reprogrammed in primordial germ cells. However, the DNA methylation dynamics at DMRs of bovine imprinted genes during early stages of development remains largely unknown. The objective of this investigation was to analyse the methylation dynamics at imprinted gene DMRs during bovine embryo development, from blastocyst stage until implantation. RESULTS: To this end, pyrosequencing technology was used to quantify DNA methylation at DMR-associated CpG dinucleotides of six imprinted bovine genes (SNRPN, MEST, IGF2R, PLAGL1, PEG10 and H19) using bisulfite-modified genomic DNA isolated from individual blastocysts (Day 7); ovoid embryos (Day 14); filamentous embryos (Day 17) and implanting conceptuses (Day 25). For all genes, the degree of DNA methylation was most variable in Day 7 blastocysts compared to later developmental stages (P < 0.05). Furthermore, mining of RNA-seq transcriptomic data and western blot analysis revealed a specific window of expression of DNA methylation machinery genes (including DNMT3A, DNMT3B, TRIM28/KAP1 and DNMT1) and proteins (DNMT3A, DNMT3A2 and DNMT3B) by bovine embryos coincident with imprint stabilization. CONCLUSION: The findings of this study suggest that the DNA methylation status of bovine DMRs might be variable during the early stages of embryonic development, possibly requiring an active period of imprint stabilization. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12861-015-0060-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-43631832015-03-19 DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development O’Doherty, Alan M Magee, David A O’Shea, Lynee C Forde, Niamh Beltman, Marijke E Mamo, Solomon Fair, Trudee BMC Dev Biol Research Article BACKGROUND: In mammals, maternal differentially methylated regions (DMRs) acquire DNA methylation during the postnatal growth stage of oogenesis, with paternal DMRs acquiring DNA methylation in the perinatal prospermatagonia. Following fusion of the male and female gametes, it is widely accepted that murine DNA methylation marks at the DMRs of imprinted genes are stable through embryogenesis and early development, until they are reprogrammed in primordial germ cells. However, the DNA methylation dynamics at DMRs of bovine imprinted genes during early stages of development remains largely unknown. The objective of this investigation was to analyse the methylation dynamics at imprinted gene DMRs during bovine embryo development, from blastocyst stage until implantation. RESULTS: To this end, pyrosequencing technology was used to quantify DNA methylation at DMR-associated CpG dinucleotides of six imprinted bovine genes (SNRPN, MEST, IGF2R, PLAGL1, PEG10 and H19) using bisulfite-modified genomic DNA isolated from individual blastocysts (Day 7); ovoid embryos (Day 14); filamentous embryos (Day 17) and implanting conceptuses (Day 25). For all genes, the degree of DNA methylation was most variable in Day 7 blastocysts compared to later developmental stages (P < 0.05). Furthermore, mining of RNA-seq transcriptomic data and western blot analysis revealed a specific window of expression of DNA methylation machinery genes (including DNMT3A, DNMT3B, TRIM28/KAP1 and DNMT1) and proteins (DNMT3A, DNMT3A2 and DNMT3B) by bovine embryos coincident with imprint stabilization. CONCLUSION: The findings of this study suggest that the DNA methylation status of bovine DMRs might be variable during the early stages of embryonic development, possibly requiring an active period of imprint stabilization. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12861-015-0060-2) contains supplementary material, which is available to authorized users. BioMed Central 2015-03-10 /pmc/articles/PMC4363183/ /pubmed/25881176 http://dx.doi.org/10.1186/s12861-015-0060-2 Text en © O'Doherty et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
O’Doherty, Alan M
Magee, David A
O’Shea, Lynee C
Forde, Niamh
Beltman, Marijke E
Mamo, Solomon
Fair, Trudee
DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development
title DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development
title_full DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development
title_fullStr DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development
title_full_unstemmed DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development
title_short DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development
title_sort dna methylation dynamics at imprinted genes during bovine pre-implantation embryo development
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4363183/
https://www.ncbi.nlm.nih.gov/pubmed/25881176
http://dx.doi.org/10.1186/s12861-015-0060-2
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