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Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection

The paternal-allele-specific methylation of the Igf2/H19 imprinting control region (ICR) is established during gametogenesis and maintained throughout development. To elucidate the requirement of the germline passage in the maintenance of the imprinting methylation, we established a system introduci...

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Autores principales: Oji, Asami, Amano, Tomojiro, Maeta, Yasuaki, Hori, Naohiro, Hatsuzawa, Kiyotaka, Sato, Kenzo, Nakanishi, Tomoko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Association for Laboratory Animal Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5682349/
https://www.ncbi.nlm.nih.gov/pubmed/28674270
http://dx.doi.org/10.1538/expanim.17-0031
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author Oji, Asami
Amano, Tomojiro
Maeta, Yasuaki
Hori, Naohiro
Hatsuzawa, Kiyotaka
Sato, Kenzo
Nakanishi, Tomoko
author_facet Oji, Asami
Amano, Tomojiro
Maeta, Yasuaki
Hori, Naohiro
Hatsuzawa, Kiyotaka
Sato, Kenzo
Nakanishi, Tomoko
author_sort Oji, Asami
collection PubMed
description The paternal-allele-specific methylation of the Igf2/H19 imprinting control region (ICR) is established during gametogenesis and maintained throughout development. To elucidate the requirement of the germline passage in the maintenance of the imprinting methylation, we established a system introducing a methylated or unmethylated ICR-containing DNA fragment (ICR-F) into the paternal or maternal genome by microinjecting into the paternal or maternal pronucleus of fertilized eggs, and traced the methylation pattern in the ICR-F. When the ICR-F was injected in a methylated form, it was demethylated approximately to half degree at blastocyst stage but was almost completely remethylated at 3 weeks of age. In the case of the unmethylated form, the ICR-F remained unmethylated at the blastocyst stage, but was almost half-methylated at 3 weeks of age. Interestingly, the paternally injected ICR-F was highly methylated compared with maternally injected ICR-F at 3 weeks of age, partially mimicking the endogenous methylation pattern. Moreover, introduction of mutations in the CTCF (CCCTC binding factor) binding sites of the ICR-F, which are known to be important for the maintenance of hypomethylated maternal ICR, induced hypermethylation of the mutated ICR-F in both paternal and maternal pronuclear injected 3-week-old mice. Our results suggest the presence of a protection-against-methylation activity of the CTCF binding site in establishing the preferential paternal methylation during post-fertilization development and the importance of germline passage in the maintenance of the parental specific methylation at H19 ICR.
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spelling pubmed-56823492017-11-16 Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection Oji, Asami Amano, Tomojiro Maeta, Yasuaki Hori, Naohiro Hatsuzawa, Kiyotaka Sato, Kenzo Nakanishi, Tomoko Exp Anim Original The paternal-allele-specific methylation of the Igf2/H19 imprinting control region (ICR) is established during gametogenesis and maintained throughout development. To elucidate the requirement of the germline passage in the maintenance of the imprinting methylation, we established a system introducing a methylated or unmethylated ICR-containing DNA fragment (ICR-F) into the paternal or maternal genome by microinjecting into the paternal or maternal pronucleus of fertilized eggs, and traced the methylation pattern in the ICR-F. When the ICR-F was injected in a methylated form, it was demethylated approximately to half degree at blastocyst stage but was almost completely remethylated at 3 weeks of age. In the case of the unmethylated form, the ICR-F remained unmethylated at the blastocyst stage, but was almost half-methylated at 3 weeks of age. Interestingly, the paternally injected ICR-F was highly methylated compared with maternally injected ICR-F at 3 weeks of age, partially mimicking the endogenous methylation pattern. Moreover, introduction of mutations in the CTCF (CCCTC binding factor) binding sites of the ICR-F, which are known to be important for the maintenance of hypomethylated maternal ICR, induced hypermethylation of the mutated ICR-F in both paternal and maternal pronuclear injected 3-week-old mice. Our results suggest the presence of a protection-against-methylation activity of the CTCF binding site in establishing the preferential paternal methylation during post-fertilization development and the importance of germline passage in the maintenance of the parental specific methylation at H19 ICR. Japanese Association for Laboratory Animal Science 2017-06-30 2017 /pmc/articles/PMC5682349/ /pubmed/28674270 http://dx.doi.org/10.1538/expanim.17-0031 Text en ©2017 Japanese Association for Laboratory Animal Science This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Original
Oji, Asami
Amano, Tomojiro
Maeta, Yasuaki
Hori, Naohiro
Hatsuzawa, Kiyotaka
Sato, Kenzo
Nakanishi, Tomoko
Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection
title Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection
title_full Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection
title_fullStr Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection
title_full_unstemmed Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection
title_short Fate of methylated/unmethylated H19 imprinting control region after paternal and maternal pronuclear injection
title_sort fate of methylated/unmethylated h19 imprinting control region after paternal and maternal pronuclear injection
topic Original
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5682349/
https://www.ncbi.nlm.nih.gov/pubmed/28674270
http://dx.doi.org/10.1538/expanim.17-0031
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