CORE-- NEUROCHEMISTRY

该计划项目的主要目标是开发可能证明 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 在AD中的应用是建立更改(改进或 可能进一步恶化)，而神经化学特征 正在接受药物治疗。它是这一神经化学的主要焦点 核心维持一个中央实验室设施，以支持 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志物的数量。这个 将提供以下服务：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(促黄体生成素和甲状腺) 促甲状腺激素[TSH]和甲状腺激素T3和T4；2)酶- 神经营养剂的联合免疫吸附试验(ELISA)服务 神经生长因子(NGF)；不同标记物的放射分析 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 C(PKC)在神经组织和神经元培养细胞中的表达； 高亲和力NGF受体蛋白(TrkA)的半定量测定 通过免疫沉淀和电泳法在两个神经组织中分离 和来自神经元培养的细胞；5)辐射监测 钙染料Fura2/AM诱导的细胞内游离钙离子 和光子计数荧光程序；6)免疫细胞化学 ChAT、环磷酸腺苷反应元件结合蛋白(CREB)、 磷酸化形式的环AMP反应元件结合蛋白(P- CREB)和线粒体原癌基因蛋白bcl2在神经细胞中的表达 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，以确定培养的神经细胞的凋亡程度 当神经元暴露于细胞侮辱，如血清剥夺时， 暴露于NMDA或β-淀粉样蛋白，。因此，神经化学的核心 将通过提供分析服务成为该项目不可或缺的一部分 用于监测选定药物在服用期间的神经化学状态 对潜力进行初步筛选和更详细的评估 治疗阿尔茨海默病及相关疾病的应用。

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.