マウス着床後初期胚における胚サイズ制御機構の解析

小鼠植入后早期胚胎大小控制机制分析

• 批准号: 22K19330
• 负责人: 佐々木 洋
• 金额: $ 4.16万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Research (Exploratory)
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-06-30 至 2025-03-31
• 项目状态: 未结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22K19330/
• 关键词: サイズ制御; 初期発生; マウス胚; Hippoシグナル

着床後初期胚のサイズ制御機構について明らかにするために、計画に沿って以下の研究を行った。（１）現象の詳細な記述：胚サイズ制御の現象は1970~80年代に行われた古典的な実験で同定されたが、その後、その現象についての解析は進んでいない。本研究では、まず、その現象を再現して、現代の技術によってその現象を正確に記述しなおすところから始める。そのために、まず胚サイズ制御が起きる時期の胎生5.5～6.5日の着床後初期胚について、サイズ制御過程を正確に記述するために、胚全体の形態、核細胞の形態、全細胞数、遺伝子発現を正確に解析するための方法を樹立した。この時期の胚は大きいため透明化しないと胚全体の細胞数がわからないが透明化は胚の形態を変えてしまうために、形態変化の少ない透明化方法を用いるとともに、透明化前後の胚の両方の画像を取得してそれぞれ目的に応じて解析に供した。また、２倍胚作製では、透明帯除去作業による軽微な毒性が発生に影響を与えがちであったが、２倍胚の作製技術がほぼ確立でき、着床前の後期胚盤胞ではエピブラスト細胞の数が２倍胚では倍になっていることが確認できた。（２）Hippoシグナルの関与の解析：胎生5.5～6.5日胚におけるHippo経路因子YAPの分布を解析した。胎生6.5日の胚では、エピブラスト細胞でYAPの核移行が見られ、Hippoシグナルが働いていないことが分かった。また、薬剤投与によりHippo経路因子LATS2キナーゼあるいはそのキナーゼ活性欠損体（LATS2-KD)を発現誘導できるトランスジェニックマウス系統を作成した。

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