NEUROTROPHIC FACTORS IN AGING AND ALZHEIMER'S DISEASE

衰老和阿尔茨海默病中的神经营养因子

• 批准号: 6267222
• 负责人: LARS OLSON
• 金额: $ 14.19万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6267222
• 关键词: Alzheimer's disease; aging; bioassay; cerebral cortex; growth factor receptors; hippocampus; human tissue; immunocytochemistry; in situ hybridization; laboratory rat; magnetic resonance imaging; molecular cloning; neurotrophic factors; nucleic acid probes; oligonucleotides; transforming growth factors

Project 5 involves a multilevel molecular genetic, cytochemical and functional approach focused on understanding the role of neurotrophic factors in the NGF and TGFbeta family in the central nervous system. In particular, we will evaluate the involvement of these factors in the maintenance of cholinergic projections and for other functions in hippocampus and cortex cerebri, as well as the possibilities to use neurotrophic factors in the NGF and TGFbeta family to reverse age-related neurodegenerative disorders. Experiments in project 5 can be subdivided as follows: (1) Cloning of new members of the TGFbeta family, emphasis on GDNF-related molecules and GDNF receptors. Accumulating evidence suggests that GDNF is an important neurotrophic factor for dopaminergic and cholinergic neurons. It is important to clone and characterize GDNF receptors, and it is very likely that GDNF, being a distant member of the TGFbeta family constitutes the first found member of a new TGFbeta sub- family. (2) Characterization of neurotrophic factors using the in vitro ganglia bioassay system; Known members of the TGFbeta family and, in particular any new GDNF-related molecules discovered in project 5 or by other scientists will be characterized in the highly specific 5-ganglia in vitro bioassay system with predictive value for in vivo effects. (3) Mapping the cellular distribution of neurotrophic factor: Using primarily in situ hybridization, but also immunohistochemistry, we will map the precise cellular localization of GDNF and any GDNF-related factors and/or GDNF receptors in the adult CNS. Additionally factors and receptors in the NGF family will be mapped in further detail. (4) Transient and permanent neurotrophic factor expression systems: GDNF, related molecules and receptors as well as NGF will be expressed by transient systems to manufacture the proteins and by permanent expression systems to be used as sources of trophic factor in collaboration with project 4. (5) Distribution and behavioral effects of neurotrophic factors injected into adult and aged normal animals: This part will evaluate in detail the spatial and temporal distribution of neurotrophic factors injected into the CSF or into brain tissue. This information is crucial for interpretation of behavioral studies which will be carried out after acute and chronic injections of neurotrophins, GDNF and novel GDNF-related factors. (6) High-resolution magnetic resonance imaging: Using new dedicated equipment enabling imaging of magnetic resonance spectroscopic information, we will monitor circulatory and metabolic changes in the brain over time in animals following intracranial injections of neurotrophic factors. (7) Functional effects of novel neurotrophic factors: Novel GDNF-related neurotrophic factors found by project 5 or by other scientists will be made using recombinant technology and used for functional studies in collaboration with project 4, (8) Studies of human post-mortem brain tissue: Postmortem brain tissue from patients with Alzheimer's disease and age-matched controls will be studied using in situ hybridization technology proven to work in such tissue to search for any possible changes in the expression of GDNF, related molecules or receptors, or neurotrophins and their receptors in Alzheimer's disease.

项目5涉及多层次的分子遗传学、细胞化学和 功能方法侧重于了解神经营养的作用， 在中枢神经系统中的NGF和TGF β家族中的因子。 在 特别是，我们将评估这些因素在 维持胆碱能投射和其他功能， 海马体和大脑皮层，以及使用的可能性， NGF和TGF β家族中的神经营养因子，以逆转年龄相关的 神经退行性疾病 项目5中的实验可以细分为 （1）TGF β家族新成员的克隆，重点是 GDNF相关分子和GDNF受体。 越来越多的证据表明 GDNF是多巴胺能神经营养因子， 胆碱能神经元 GDNF的克隆和鉴定具有重要意义 受体，很可能GDNF，作为一个遥远的成员， TGF β家族是第一个发现的新TGF β亚家族成员， 家人 (2)神经营养因子的体外表征 神经节生物测定系统; TGF β家族的已知成员， 特别是在项目5中发现的任何新的GDNF相关分子， 其他科学家将在高度特异性的5-神经节的特点， 体外生物测定系统，具有体内效应的预测值。 （三） 定位神经营养因子的细胞分布：主要使用 原位杂交和免疫组织化学，我们将绘制 GDNF和任何GDNF相关因子的精确细胞定位和/或 GDNF受体在成年人中枢神经系统。 此外，在细胞中的因子和受体 将进一步详细绘制NGF家族。 (4)暂时性和永久性 神经营养因子表达系统：GDNF，相关分子和 受体以及NGF将通过瞬时系统表达， 生产蛋白质，并通过永久表达系统用作 与项目4合作研究营养因子的来源。 （五） 神经营养因子在大鼠脑内的分布及行为学效应 成年和老年正常动物：本部分将详细评价 神经营养因子的时空分布 脑脊液或脑组织。 这些信息对于解释至关重要 行为研究，将在急性和慢性 注射神经营养因子、GDNF和新的GDNF相关因子。 （六） 高分辨率磁共振成像：使用新的专用设备 使磁共振光谱信息成像，我们将 监测动物大脑随时间的循环和代谢变化 在颅内注射神经营养因子后 (7)功能 新型神经营养因子的作用：新型GDNF相关神经营养因子 项目5或其他科学家发现的因素将使用 重组技术和用于功能研究的合作 项目4，（8）人类死后脑组织的研究：死后 阿尔茨海默病患者的脑组织和年龄匹配的 将使用原位杂交技术对对照进行研究， 在这样的组织中工作，以寻找任何可能的表达变化， GDNF，相关分子或受体，或神经营养因子及其受体 老年痴呆症