Analysis of the prion mechanism using immortalized prionless neuronal cell

利用永生化无朊病毒神经元细胞分析朊病毒机制

• 批准号: 06454128
• 负责人: ONODERA Takashi
• 金额: $ 4.61万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454128/
• 关键词: prion; gene targeting; spongiform encephalopathy; scrapie; neurogenedegenerative diseases; slow virus; kuru; cell immortalization; スクレイビ-; 海綿状脳症; 痴呆症

Prion is a cellular protein (PrPc) which has been implicated in transmissible neurodegenerative disease This protein is found on the outer surfaces of neurons of the brain. Although the PrPc function has still not been completely clarified, it seems to be physiologically important because its gene has been found in all mammal examined so far. The establishment of progenitor cell lines from Prn-p gene-target knock out mice embryos and from the wild type would be a valuable model system to clarify if PrPc is related with putative differences in the development of those two cell system and also to verify if there is presence of compensatory gene control mechanisms early in embryogenesis.Mice homozogous for disrupted Prn-p genes have been generated (Bueler et al. 1987).From this type of gene-targeted mice, hippocampal cells have been immortalized and clones were obtatined. The hippocampus has been dissected from brain of mouse embryos at embryonic day 14, as the PrPc is predominantly a neuronal protein, presented in a high proportion of hippocampus nerve cell, and that the neurogenesis in vivo are prefferentially occuring at early development.Twenty eight clones were obtained and has been characterized as prionless and neuronal origin.

朊病毒是一种细胞蛋白 (PrPc)，与传染性神经退行性疾病有关。这种蛋白存在于大脑神经元的外表面。尽管PrPc的功能尚未完全阐明，但它似乎具有重要的生理意义，因为迄今为止在所有检查的哺乳动物中都发现了它的基因。从 Prn-p 基因靶向敲除小鼠胚胎和野生型中建立祖细胞系将是一个有价值的模型系统，可以阐明 PrPc 是否与这两个细胞系统发育中的推定差异相关，并验证是否与这两种细胞系统的发育差异相关。在胚胎发生的早期就存在补偿性基因控制机制。已经产生了与破坏的 Prn-p 基因同源的小鼠（Bueler 等人，1987）。从这种类型的基因靶向小鼠中，海马细胞已永生化并获得克隆。在胚胎第 14 天从小鼠胚胎的大脑中解剖出海马，因为 PrPc 主要是一种神经元蛋白，存在于高比例的海马神经细胞中，并且体内神经发生优先发生在早期发育时。 28 个克隆获得并被表征为无朊病毒和神经元来源。

项目成果

佐伯圭一ら: "Identification of a promotor region in the rat prion protein gene." Biochem.Biophs.Res.Comm.120(in press). (1995)

Keiichi Saeki 等人：“大鼠朊病毒蛋白基因启动子区域的鉴定”。Biochem.Biophs.Res.Comm.120（出版中）。

Onodera T.,他: "Diversity of clinical sign in natural scrapie cases in Japan." Jpn. Agri. Res. Quart.28. 59-61 (1994)

Onodera T. 等人：“日本自然瘙痒病病例的多样性”，Agri Res.28（1994 年）。

佐伯 圭一,他: "ラット不死化神経細胞株およびラットにおけるプリオン蛋白のcDNAのクローニングと遺伝子発現の解析" 家畜生化学. 31. 37-43 (1994)

Keiichi Saeki 等：“永生化大鼠神经元细胞系和大鼠中朊病毒蛋白 cDNA 的克隆和基因表达分析”Livestock Biochemistry 31. 37-43 (1994)。

Takashi Onodera: "Prion (in Japanese)" Biochemistry (in Japanese). 67. 1249 (1995)

Takashi Onodera：“朊病毒（日语）”生物化学（日语）。

丹治邦和ら: "Analysis of PrP^C mRNA by in situ hybridization in brain, placenta, uterus and testis of rat." Intervirology. 38. (in press) (1995)

Kunikazu Tanji 等人：“通过原位杂交对大鼠脑、胎盘、子宫和睾丸中的 PrP^C mRNA 进行分析。Intervirology”（正在出版）（1995 年）。