EXOGENOUS INDUCTION OF ALZHEIMER A?-PATHOLOGY IN TRANSGENIC MICE

转基因小鼠中阿尔茨海默病 A 型病理的外源诱导

• 批准号: 7715846
• 负责人: LARY C WALKER
• 金额: $ 2.85万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7715846
• 关键词: APP-PS1; Alzheimer' s Disease; Animals; Antibodies; Brain; Characteristics; Computer Retrieval of Information on Scientific Projects Database; Confocal Microscopy; Contralateral; Deposition; Enzyme-Linked Immunosorbent Assay; Funding; Grant; Hippocampus (Brain); Human; Immunofluorescence Immunologic; Immunoprecipitation; Injection of therapeutic agent; Institution; MALDI-TOF Mass Spectrometry; Mus; Pathology; Patients; Pattern; Peptides; Population; Primates; Research; Research Personnel; Resistance; Resources; Saimiri; Seeds; Source; Testing; Transgenic Mice; United States National Institutes of Health; aged; internal control

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have examined the quantitative and qualitative characteristics of A¿ in humans with AD and in squirrel monkeys (Saimiri sciureus), a closely related, but AD-resistant, primate species. By ELISA, we found that cortical A¿ levels in aged squirrel monkeys can equal or exceed the levels in AD brain. Using immunoprecipitation and MALDI-TOF mass spectrometry, we have also shown that soluble and insoluble A¿ peptide populations are highly similar in squirrel monkeys and humans with AD. To test the hypothesis that A¿-multimers in the two species may assume pathogenically dissimilar forms, we then examined the ability of A¿-rich cortical extracts from squirrel monkeys and AD patients to induce, or seed, A¿ deposition in APP/PS1 transgenic mice. Dilute cortical extracts were injected unilaterally into the hippocampus of APP/PS1 mice, prior to the endogenous deposition of A¿ in that region. Sham PBS injections in the contralateral hemisphere served as internal controls. After a 4 month incubation period, the mouse brains were immunostained with antibodies to the A¿ peptide. A¿ deposition was enhanced in the extract-injected hemisphere of all experimental animals. With double immunofluorescence and confocal microscopy, we identified cellular and parenchymal patterns of seeded A¿ deposition unique to AD and squirrel monkey extracts.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们研究了患有 AD 的人类和松鼠猴 (Saimiri sciureus)（一种密切相关但具有 AD 抗性的灵长类物种）中 A¿ 的定量和定性特征。 通过ELISA，我们发现老年松鼠猴的皮质A¿水平可以等于或超过AD大脑中的水平。 使用免疫沉淀和 MALDI-TOF 质谱法，我们还表明，松鼠猴和患有 AD 的人类中可溶性和不溶性 A¿ 肽群高度相似。 为了检验这两个物种中的 A 多聚体可能呈现致病性不同形式的假设，我们随后检查了松鼠猴和 AD 患者富含 A 的皮质提取物在 APP/PS1 转基因小鼠中诱导或播种 A 沉积的能力。 在 A¿ 内源性沉积到该区域之前，将稀释的皮质提取物单侧注射到 APP/PS1 小鼠的海马体中。 对侧半球假 PBS 注射作为内部对照。 经过 4 个月的潜伏期后，用 A¿ 肽的抗体对小鼠大脑进行免疫染色。 所有实验动物注射提取物的半球中的沉积均得到增强。 通过双重免疫荧光和共聚焦显微镜，我们鉴定了 AD 和松鼠猴提取物特有的种子 A 沉积的细胞和实质模式。