TRANSGENIC CORE

转基因核心

• 批准号: 6920483
• 负责人: EFRAT LEVY
• 金额: $ 20.28万
• 依托单位: NATHAN S. KLINE INSTITUTE FOR PSYCH RES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6920483
• 关键词: Alzheimer' s disease; Downs syndrome; animal breeding; animal colony; autophagy; biomedical facility; cellular pathology; endocytosis; genetic models; genetically modified animals; genotype; laboratory mouse; lysosomes; model design /development; molecular pathology; neuropathology

The use of transgenic animals is a powerful and informative approach that has been successfully undertaken by the core PL for over five years to study neurodegenerative diseases. The purpose of the transgenic core is to extend production of mouse models of Down's syndrome (Ts21), Alzheimer's disease (AD), and other neurodegenerative disorders and to generate novel crosses between existing models, including mice showing endocytic-autophagic-lysosomal system (EALS) pathology. The Ts65Dn mouse model develops endosomal abnormalities similar to those in AD and is used by all the projects. In order to modulate the endocytic pathology in these mice, they will be crossed with mice expressing human APP, null for mouse APP, or APP-secretase deficient. In human AD, inheritance of the ApoE D4 allele exacerbated endosomal alteration, which will be further examined in Ts65Dn crossed to humanized ApoE mice. The role for APP triplication in AD-related endosome dysfunction in these mice will be explored by single-cell DMA microarray analyses using Ts65Dn and Ts65Dn/APP null crosses. The second Project has identified a key role for presenilin in modulating autophagy, which will be examined in PS1 hypomorphic and a mutant PS1 transgenics and further explored in crosses with Ts65Dn and an amyloid depositing mouse. A transgenic overexpressing LC3, a regulator of autophagy, will be used as a model of autophagy dysregulation. The third Project will define the neuroprotective role that cystatin C plays in of AD-related EALS dysfunction using these same mouse models. Modulation of cystatin C levels will be obtained by crossing these models with either cystatin C transgenic or knockout mice. The potential for redundancy between cystatin B, another cysteine protease inhibitor, and cystatin C will be investigated by using cystatin B knockout mice. The core leader has extensive experience managing a large mouse colony with complex breeding schemes, and will be able to co-ordinate animal breeding, genotyping, and tracking of animals.

使用转基因动物是一种强大且信息丰富的方法，核心 PL 已成功采用该方法五年多来研究神经退行性疾病。转基因核心的目的是扩大唐氏综合症（Ts21）、阿尔茨海默病（AD）和其他神经退行性疾病小鼠模型的生产，并在现有模型之间产生新的杂交，包括显示内吞自噬溶酶体系统（EALS）的小鼠）病理学。 Ts65Dn 小鼠模型开发 内体异常与 AD 中的异常相似，并被所有项目使用。为了调节这些小鼠的内吞病理学，将它们与表达人APP、小鼠APP无效或APP分泌​​酶缺陷的小鼠杂交。在人类 AD 中，ApoE D4 等位基因的遗传加剧了内体改变，这将在与人源化 ApoE 小鼠杂交的 Ts65Dn 中进一步检查。将通过使用 Ts65Dn 和 Ts65Dn/APP 无效杂交的单细胞 DMA 微阵列分析来探索 APP 三倍体在这些小鼠 AD 相关内体功能障碍中的作用。第二个项目已经确定了早老素在调节自噬中的关键作用，将在 PS1 低等态和突变 PS1 转基因中进行检查，并在与 Ts65Dn 和淀粉样蛋白沉积小鼠的杂交中进一步探索。一个 转基因过表达LC3（一种自噬调节因子）将被用作自噬模型 失调。第三个项目将使用这些相同的小鼠模型来定义半胱氨酸蛋白酶抑制剂 C 在 AD 相关的 EALS 功能障碍中所发挥的神经保护作用。通过将这些模型与半胱氨酸蛋白酶抑制剂 C 转基因或敲除小鼠杂交，可以获得半胱氨酸蛋白酶抑制剂 C 水平的调节。半胱氨酸蛋白酶抑制剂 B（另一种半胱氨酸蛋白酶抑制剂）与半胱氨酸蛋白酶抑制剂 C 之间的冗余潜力将通过使用半胱氨酸蛋白酶抑制剂 B 敲除小鼠进行研究。核心领导者拥有管理大型小鼠群体和复杂育种计划的丰富经验，并且能够协调动物育种、基因分型和动物追踪。