CORE-- NEUROCHEMISTRY

核心——神经化学

• 批准号: 6098416
• 负责人: William J Millard
• 金额: $ 15.44万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6098416
• 关键词: Alzheimer's disease; aging; binding proteins; biomarker; biomedical facility; brain cell; cAMP response element binding protein; calcium flux; choline acetyltransferase; enzyme activity; enzyme linked immunosorbent assay; growth factor receptors; immunocytochemistry; immunoprecipitation; neurochemistry; neurons; neuropeptides; neurotransmitter metabolism; neurotransmitters; neurotrophic factors; oncoproteins; pituitary hormones; protein kinase C; radioimmunoassay; radiotracer; tissue /cell culture

The major goal of this program project is to develop drugs that may prove beneficial to Alzheimer's disease (AD) patients. Paramount to the establishment of a drug or class of drugs that may have therapeutic applications in AD is the establishment of alterations (improvements or possibly further deterioration) in the neurochemical profile while undergoing drug therapy. It is the primary focus of this Neurochemistry Core to maintain a central laboratory facility which will support the various projects outlined in the program by providing assay service for a number of hormonal, cholinergic and peptidergic neurochemical markers. The services provided will be as follows; 1) radioimmunoassay (RIA) support for the various estrogens (17beta-estradiol, 17alpha-estradiol, estrone, estriol), anterior pituitary hormones (luteinizing hormone [LH] and thyroid stimulating hormone [TSH] and the thyroid hormones T3 and T4; 2) enzyme- linked immunoabsorbant assay (ELISA) service for the neurotrophic agent nerve growth factor (NGF); 3). radiometric analysis of various markers of cholinergic function (high affinity choline uptake [HACU], acetylcholine release and choline acetyltransferase [ChAT] activity) and protein kinase C (PKC) in both neural tissue and cells from neuron-enriched cultures; 4) semiquantitative measurement of high affinity NGF receptor protein (trkA) by immunoprecipitation and electrophoretic separation in both neural tissue and cells from neuron-enriched cultures; 5) radiometric monitoring of intracellular ionized (free) calcium [Ca++]i by using calcium dye Fura2/AM and photon counting spectrofluorometric procedures; 6) immunocytochemical analysis of ChAT, cyclic AMP response element binding protein (CREB), phosphorylated form of the cyclic AMP response element binding protein (P- CREB) and mitochondrial proto-oncogene protein bcl-2 levels in neural tissue and 7) in situ end-labeling of DNA oligonucleotides in brain tissue and cultured neurons to determine the extent of apoptosis in cultured neurons when exposed to cellular insults such as serum deprivation, exposure to NMDA or beta-amyloid protein,. Thus, the Neurochemistry Core will be integral to this program project by providing analytical service for the monitoring of neurochemical status while selected drugs undergo both preliminary screening and more detailed evaluation for potential therapeutic uses in the treatment of AD and related disorders.

该计划项目的主要目标是开发可以证明的药物 对阿尔茨海默病（AD）患者有益。 最重要的是 建立一种或一类可能具有治疗作用的药物 AD 中的应用程序是建立变更（改进或 神经化学特征可能进一步恶化，同时 正在接受药物治疗。 这是神经化学的主要焦点 核心是维持一个中央实验室设施，以支持 该计划中概述的各种项目通过提供化验服务 激素、胆碱能和肽能神经化学标记物的数量。 这 提供的服务如下； 1) 放射免疫分析（RIA）支持 对于各种雌激素（17β-雌二醇、17α-雌二醇、雌酮、 雌三醇）、垂体前叶激素（黄体生成素 [LH] 和甲状腺 刺激激素 [TSH] 以及甲状腺激素 T3 和 T4； 2) 酶- 神经营养剂的连锁免疫吸附测定 (ELISA) 服务 神经生长因子（NGF）； 3）。 各种标记物的辐射分析 胆碱能功能（高亲和力胆碱摄取 [HACU]、乙酰胆碱 释放和胆碱乙酰转移酶 [ChAT] 活性）和蛋白激酶 神经组织和神经元富集培养物细胞中的 C (PKC)； 4） 高亲和力 NGF 受体蛋白 (trkA) 的半定量测量 通过免疫沉淀和电泳分离两种神经组织 以及来自富含神经元的培养物的细胞； 5）辐射监测 使用钙染料 Fura2/AM 检测细胞内离子（游离）钙 [Ca++]i 和光子计数荧光光谱程序； 6) 免疫细胞化学 ChAT、环AMP反应元件结合蛋白（CREB）的分析， 环 AMP 反应元件结合蛋白的磷酸化形式（P- CREB）和线粒体原癌基因蛋白 bcl-2 水平在神经 组织和 7) 脑组织中 DNA 寡核苷酸的原位末端标记 并培养神经元以确定培养物中细胞凋亡的程度 当神经元受到细胞损伤（例如血清剥夺）时， 暴露于 NMDA 或 β-淀粉样蛋白。 因此，神经化学核心 通过提供分析服务将成为该计划项目的一部分 用于在选定的药物接受治疗时监测神经化学状态 初步筛选和更详细的潜力评估 在治疗 AD 和相关疾病中的治疗用途。