NEUROTROPHIC FACTORS IN AGING AND ALZHEIMER'S DISEASE

项目5涉及多水平的分子遗传、细胞化学和 功能方法侧重于了解神经营养的作用 中枢神经系统中神经生长因子和转化生长因子β家族的因子。在……里面 特别是，我们将评估这些因素在 胆碱能投射的维持和其他功能 海马体和大脑皮质，以及使用的可能性 神经生长因子和转化生长因子β家族中的神经营养因子逆转年龄相关性 神经退行性疾病。项目5中的实验可以细分为 如下：(1)克隆TGFbeta家族的新成员，重点是 神经营养因子相关分子和神经营养因子受体。越来越多的证据表明 GDNF是一种重要的神经营养因子，对多巴胺能和 胆碱能神经元。GDNF的克隆和鉴定具有重要意义 受体，很可能是GDNF，作为 TGFbeta家族构成了新的TGFbeta亚家族中的第一个成员 一家人。(2)神经营养因子的体外鉴定 神经节生物测定系统；TGFbeta家族的已知成员，以及，在 特别是在项目5中发现的任何与GDNF相关的新分子 其他科学家将对高度特异的5-神经节进行研究 具有体内效应预测价值的体外生物测定系统。(3) 绘制神经营养因子的细胞分布图：主要使用 原位杂交，还有免疫组织化学，我们会将图谱 胶质细胞源性神经营养因子及其相关因子和/或的精确细胞定位 成体中枢神经系统的GDNF受体。此外，血管内皮细胞的因子和受体 NGF家族将被更详细地绘制。(四)暂时性和永久性 神经营养因子表达系统：GDNF、相关分子和 受体和NGF将通过瞬时系统表达，以 制造蛋白质并通过永久表达系统用作 与项目4合作的营养因子来源。(5) 脑内注射神经营养因子的分布和行为效应 成年和老年正常动物：这一部分将详细评估 脑内注射神经营养因子的时空分布 脑脊液或进入脑组织。这一信息对口译至关重要。 将在急性和慢性之后进行的行为研究 神经营养因子、胶质细胞源性神经营养因子及新型神经营养因子相关因子的注射。(6) 高分辨率磁共振成像：使用新的专用设备 启用磁共振光谱信息的成像，我们将 监测动物脑内循环和代谢随时间的变化 在脑部注射神经营养因子之后。(7)功能 新型神经营养因子的作用：与神经营养因子相关的新型神经营养 项目5或其他科学家发现的因子将使用 重组技术，用于合作进行功能研究 项目4，(8)人死后脑组织的研究：死后 阿尔茨海默病患者和年龄匹配患者的脑组织 将使用原位杂交技术进行对照研究，证明 在这样的组织中工作，寻找任何可能的表达变化 神经营养因子、相关分子或受体，或神经营养因子及其受体 阿尔茨海默氏症。

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.