Analysis of Epigenome Marks and Transcriptome in the Germ Line by the Next Generation Sequencer

2014 Fiscal Year Final Research Report

• Project/Area Number: 22228004
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Applied animal science
• Research Institution: Tokyo University of Agriculture
• Principal Investigator: KONO Tomohiro 東京農業大学, 応用生物科学部, 教授 (80153485)
• Co-Investigator(Kenkyū-buntansha): SOTOMARU Yusuke 広島大学, 自然科学研究支援センター, 教授 (90309352) ; SUZUKI Yutaka 東京大学, 新領域形成研究科, 教授 (40323646)
• Research Collaborator: KOBAYASHI Hisato 東京農業大学, 応用生物科学部・生物資源ゲノム解析センター, 准教授 (70632727)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Keywords: DNA メチローム / 生殖系列 / 次世代シークエンサー / トランスクリプトーム / 始原生殖細胞 / 生殖細胞 / リプログラミング

Outline of Final Research Achievements

To clarify DNA methylation reprogramming and transcriptome in the whole female and male germline lineage in mice, a comprehensive DNA methylation analysis for all cytosine was conducted using a next-generation sequencer. DNA methylation was 30% and 80% in the oocytes and sperm, respectively. After fertilization the level decreased and reached to 18% at the blastocysts, however, in the E7.5 epiblast, the DNA methylation was quickly increased to 73%. In the primordial germ cells, which are differentiated at this stage, demethylation occurred rapidly, and the methylation level deceased to less than 5% in both sexes at E13.5. Subsequently, female- and male-specific DNA methylomes were established independently in the sparmatogonium before birth and during oocyte growth period, respectively. Transcriptome analysis revealed that DNA methylation is a critical regulator of gene expression in germline cells. This is the first report of a complete DNA methylome map of the whole germline in mice.

Free Research Field

生殖科学