CORE-- NEUROCHEMISTRY

核心——神经化学

• 批准号: 6360491
• 负责人: William J Millard
• 金额: $ 15.3万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6360491
• 关键词: Alzheimer's disease; BCL2 gene /protein; biomarker; biomedical facility; brain cell; cAMP response element binding protein; calcium flux; choline acetyltransferase; drug design /synthesis /production; estrogen analog; immunocytochemistry; immunoprecipitation; neurochemistry; neurotrophic factors; protein kinase C; radioimmunoassay; 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.

该项目的主要目标是开发药物， 对阿尔茨海默病（AD）患者有益。 派拉蒙到 确定一种或一类可能具有治疗作用的药物 在AD中的应用是建立改建（改进或 可能进一步恶化），而 正在接受药物治疗 这是神经化学的主要焦点 核心维持一个中心实验室设施， 该计划中概述的各种项目，通过为 激素、胆碱能和肽能神经化学标记物的数量。 的 提供的服务如下：1）放射免疫分析支助 对于各种雌激素（17 β-雌二醇，17 α-雌二醇，雌酮， 雌三醇），垂体前叶激素（黄体生成素[LH]和甲状腺激素 刺激激素[TSH]和甲状腺激素T3和T4; 2）酶- 用于神经营养剂的连接免疫吸附测定（ELISA）服务 神经生长因子（NGF）; 放射性分析的各种标志物 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 C（PKC）在神经组织和来自神经元富集培养物的细胞中; 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， 和来自神经元富集培养物的细胞; 5） 用钙染料Fura 2/AM测定细胞内游离钙离子浓度 和光子计数荧光光谱法; 6）免疫细胞化学 ChAT，环AMP反应元件结合蛋白（CREB）， 磷酸化形式的环AMP反应元件结合蛋白（P- 神经元细胞中CREB）和线粒体原癌基因蛋白bcl-2的水平 组织和7）脑组织中DNA寡核苷酸的原位末端标记 和培养的神经元，以确定培养的细胞凋亡的程度。 当神经元暴露于细胞损伤如血清剥夺时， 暴露于NMDA或β-淀粉样蛋白。 因此，神经化学核心 将通过提供分析服务成为该计划项目的组成部分 用于在选定药物进行治疗时监测神经化学状态， 初步筛选和更详细的评估， 在治疗AD和相关病症中的治疗用途。