Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth

Akihiko Sakashita, Hisato Kobayashi, Takuya Wakai, Yusuke Sotomaru, Kenichiro Hata, Tomohiro Kono

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Recent studies of the demethylation process in murine zygotes have shown that 5-methylcytosine (5mC) is first converted into 5-hydroxymethylcytosine (5hmC) or further-oxidized cytosines in the paternal genome by the maternal ten-eleven translocation 3 (TET3) enzyme. This process is crucial for normal embryogenesis, and our aim was to elucidate the effect of Tet3 on the maternal genome during female germ-line development. Immunofluorescence analysis showed that 5hmC was clearly present in fully grown oocytes but not in nongrowing and early growth-stage oocytes. The 5hmC in the maternal genome was clearly detectable in DNA methyltransferase 3-like enzyme (Dnmt3L)-null oocytes and their fertilized zygotes, although Dnmt3L is essential for DNA methylation in oocytes. An analysis using an enzyme digestion-based method showed that 5hmC was present in LTR retrotransposons from the late growth period of oocytes. Quantitative RT-PCR analysis showed that Tet3 expression was enhanced during oocyte growth and exhibited an approximately 40-fold increase between nongrowing and fully grown oocytes. Our results show that 5hmC is generated since the oocyte growth stage, accompanied by up-regulation of Tet3; 5hmC is located mainly in LTR retrotransposons, indicating that 5hmC generated in growth-stage oocytes is responsible for genomewide demethylation after fertilization.

Original languageEnglish
Pages (from-to)629-636
Number of pages8
JournalGenes to Cells
Volume19
Issue number8
DOIs
Publication statusPublished - 2014
Externally publishedYes

Fingerprint

Oocytes
Growth
Retroelements
Zygote
Mothers
Methyltransferases
Genome
Enzymes
5-Methylcytosine
5-hydroxymethylcytosine
Cytosine
DNA
DNA Methylation
Fertilization
Germ Cells
Embryonic Development
Fluorescent Antibody Technique
Digestion
Up-Regulation
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Genetics
  • Cell Biology
  • Medicine(all)

Cite this

Sakashita, A., Kobayashi, H., Wakai, T., Sotomaru, Y., Hata, K., & Kono, T. (2014). Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth. Genes to Cells, 19(8), 629-636. https://doi.org/10.1111/gtc.12164

Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth. / Sakashita, Akihiko; Kobayashi, Hisato; Wakai, Takuya; Sotomaru, Yusuke; Hata, Kenichiro; Kono, Tomohiro.

In: Genes to Cells, Vol. 19, No. 8, 2014, p. 629-636.

Research output: Contribution to journalArticle

Sakashita, A, Kobayashi, H, Wakai, T, Sotomaru, Y, Hata, K & Kono, T 2014, 'Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth', Genes to Cells, vol. 19, no. 8, pp. 629-636. https://doi.org/10.1111/gtc.12164
Sakashita, Akihiko ; Kobayashi, Hisato ; Wakai, Takuya ; Sotomaru, Yusuke ; Hata, Kenichiro ; Kono, Tomohiro. / Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth. In: Genes to Cells. 2014 ; Vol. 19, No. 8. pp. 629-636.
@article{60f0b74620b148e0ad518d7d71aa1064,
title = "Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth",
abstract = "Recent studies of the demethylation process in murine zygotes have shown that 5-methylcytosine (5mC) is first converted into 5-hydroxymethylcytosine (5hmC) or further-oxidized cytosines in the paternal genome by the maternal ten-eleven translocation 3 (TET3) enzyme. This process is crucial for normal embryogenesis, and our aim was to elucidate the effect of Tet3 on the maternal genome during female germ-line development. Immunofluorescence analysis showed that 5hmC was clearly present in fully grown oocytes but not in nongrowing and early growth-stage oocytes. The 5hmC in the maternal genome was clearly detectable in DNA methyltransferase 3-like enzyme (Dnmt3L)-null oocytes and their fertilized zygotes, although Dnmt3L is essential for DNA methylation in oocytes. An analysis using an enzyme digestion-based method showed that 5hmC was present in LTR retrotransposons from the late growth period of oocytes. Quantitative RT-PCR analysis showed that Tet3 expression was enhanced during oocyte growth and exhibited an approximately 40-fold increase between nongrowing and fully grown oocytes. Our results show that 5hmC is generated since the oocyte growth stage, accompanied by up-regulation of Tet3; 5hmC is located mainly in LTR retrotransposons, indicating that 5hmC generated in growth-stage oocytes is responsible for genomewide demethylation after fertilization.",
author = "Akihiko Sakashita and Hisato Kobayashi and Takuya Wakai and Yusuke Sotomaru and Kenichiro Hata and Tomohiro Kono",
year = "2014",
doi = "10.1111/gtc.12164",
language = "English",
volume = "19",
pages = "629--636",
journal = "Genes to Cells",
issn = "1356-9597",
publisher = "Wiley-Blackwell",
number = "8",

}

TY - JOUR

T1 - Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth

AU - Sakashita, Akihiko

AU - Kobayashi, Hisato

AU - Wakai, Takuya

AU - Sotomaru, Yusuke

AU - Hata, Kenichiro

AU - Kono, Tomohiro

PY - 2014

Y1 - 2014

N2 - Recent studies of the demethylation process in murine zygotes have shown that 5-methylcytosine (5mC) is first converted into 5-hydroxymethylcytosine (5hmC) or further-oxidized cytosines in the paternal genome by the maternal ten-eleven translocation 3 (TET3) enzyme. This process is crucial for normal embryogenesis, and our aim was to elucidate the effect of Tet3 on the maternal genome during female germ-line development. Immunofluorescence analysis showed that 5hmC was clearly present in fully grown oocytes but not in nongrowing and early growth-stage oocytes. The 5hmC in the maternal genome was clearly detectable in DNA methyltransferase 3-like enzyme (Dnmt3L)-null oocytes and their fertilized zygotes, although Dnmt3L is essential for DNA methylation in oocytes. An analysis using an enzyme digestion-based method showed that 5hmC was present in LTR retrotransposons from the late growth period of oocytes. Quantitative RT-PCR analysis showed that Tet3 expression was enhanced during oocyte growth and exhibited an approximately 40-fold increase between nongrowing and fully grown oocytes. Our results show that 5hmC is generated since the oocyte growth stage, accompanied by up-regulation of Tet3; 5hmC is located mainly in LTR retrotransposons, indicating that 5hmC generated in growth-stage oocytes is responsible for genomewide demethylation after fertilization.

AB - Recent studies of the demethylation process in murine zygotes have shown that 5-methylcytosine (5mC) is first converted into 5-hydroxymethylcytosine (5hmC) or further-oxidized cytosines in the paternal genome by the maternal ten-eleven translocation 3 (TET3) enzyme. This process is crucial for normal embryogenesis, and our aim was to elucidate the effect of Tet3 on the maternal genome during female germ-line development. Immunofluorescence analysis showed that 5hmC was clearly present in fully grown oocytes but not in nongrowing and early growth-stage oocytes. The 5hmC in the maternal genome was clearly detectable in DNA methyltransferase 3-like enzyme (Dnmt3L)-null oocytes and their fertilized zygotes, although Dnmt3L is essential for DNA methylation in oocytes. An analysis using an enzyme digestion-based method showed that 5hmC was present in LTR retrotransposons from the late growth period of oocytes. Quantitative RT-PCR analysis showed that Tet3 expression was enhanced during oocyte growth and exhibited an approximately 40-fold increase between nongrowing and fully grown oocytes. Our results show that 5hmC is generated since the oocyte growth stage, accompanied by up-regulation of Tet3; 5hmC is located mainly in LTR retrotransposons, indicating that 5hmC generated in growth-stage oocytes is responsible for genomewide demethylation after fertilization.

UR - http://www.scopus.com/inward/record.url?scp=84925351861&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84925351861&partnerID=8YFLogxK

U2 - 10.1111/gtc.12164

DO - 10.1111/gtc.12164

M3 - Article

VL - 19

SP - 629

EP - 636

JO - Genes to Cells

JF - Genes to Cells

SN - 1356-9597

IS - 8

ER -