Generation and application of mtDNA knockout mice as models for mitochondrial diseases

线粒体DNA敲除小鼠线粒体疾病模型的产生及应用

• 批准号: 10358018
• 负责人: HAYASHI Jun-ichi
• 金额: $ 19.19万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10358018/
• 关键词: Mouse mtDNA; mtDNA Mutation; Mitochondrial transfer; Disease model mice; mitochondrial tRNA; mtDNA突然変異; ミトコンドリアVRNA; マウス; ミトコンドリアDNA; 遺伝子導入; 突然変異; ミトコンドリア遺伝子病; 病態モデルマウス; ミトコンドリアDNAノックアウトマウス; マウス受精卵; マイクロインジェクション法; 呼吸欠損株

Mice possessing pathogenic mutant mitochondrial DNA (mtDNA) would provide ideal systems for studying exactly how mutant mtDNAs are transmitted and distributed in tissues resulting in expression of mitochondrial diseases, but no effective procedures are available for their generation. Isolation of mtDNA-less (ρ^0) mouse cells enabled us to trap mouse mutant mtDNAs that had accumulated in somatic tissues into mtDNA repopulated ρ^0 cells (cybrids). We could isolate respiration-deficient cybrids with a deletion mutant mtDNA and introduce it into fertilized eggs. The mutant mtDNA was transmitted maternally, and its accumulation induced mitochondrial dysfunction in various tissues. Moreover, most of these mice unexpectedly died as a consequence of renal failure, suggesting the involvement of mtDNA mutations in the pathogeneses of new diseases. Escape ofJhe tissues with up to 90 % mutant mtDNA from expression of disease phenotypes was enabled by the extensive intermitochondrial interaction.

具有致病性突变线粒体DNA（mtDNA）的小鼠将为研究突变mtDNA在组织中的传播和分布提供理想的系统，从而导致线粒体疾病的表达，但没有有效的方法可用于其产生。分离出缺失mtDNA的（ρ^0）小鼠细胞使我们能够捕获在体细胞组织中积累的小鼠突变mtDNA，使其进入mtDNA重新填充的ρ^0细胞（胞质杂交体）。我们可以分离出带有缺失突变mtDNA的呼吸缺陷型胞质杂种，并将其导入受精卵。突变的线粒体DNA是母系传递的，其积累诱导各种组织中的线粒体功能障碍。此外，这些小鼠中的大多数意外地死于肾衰竭，这表明mtDNA突变参与了新疾病的发病机制。广泛的线粒体相互作用使具有高达90%突变mtDNA的组织能够逃避疾病表型的表达。

项目成果

Ito,S.: "Functional integrity of mitochondrial genomes in human platelets and autopsied brain tissues from aged patients with Alzheimer's disease."Proc.Natl.Acad.Sci.USA. 96. 2099-2103 (1999)

Ito,S.：“人类血小板和老年阿尔茨海默病患者尸检脑组织中线粒体基因组的功能完整性。”Proc.Natl.Acad.Sci.USA。

Kazuto Nakada: "Mitochondria-specific system preventing expression of disease phenotypes by mutant mtDNA"Cell Technology. 20. 1552-1565 (2001)

Kazuto Nakada：“线粒体特异性系统通过突变 mtDNA 阻止疾病表型的表达”细胞技术。

Kazuto Nakada: "Inter-mitochondrial complementation : mitochondria-specific system preventing mice from expression of disease phenotypes by mutant mtDNA"Nature Med.. 7. 934-939 (2001)

Kazuto Nakada：“线粒体间互补：线粒体特异性系统通过突变 mtDNA 防止小鼠表达疾病表型”Nature Med.. 7. 934-939 (2001)

Masato Tawata: "A new mtDNA mutation at 14577 T/C is probably a major pathogenic mutation of matemally inherited type 2 diabetes"Diabetes. 49. 1269-1272 (2000)

Masato Tawata：“14577 T/C 处的新 mtDNA 突变可能是母系遗传的 2 型糖尿病的主要致病突变”糖尿病。

Hiroshi Shitara: "Non-invasive visualization of sperm mitochondrial behavior in trausgenic mice with introduced green fluorescent protein (GFP)"FEBS Lett.. 500. 7-11 (2001)

Hiroshi Shitara：“引入绿色荧光蛋白 (GFP) 的创伤性小鼠精子线粒体行为的非侵入性可视化”FEBS Lett.. 500. 7-11 (2001)