BRAIN GLUCOSE METABOLISM BY 13C NMR

申请人摘要：乙醇对谷氨酸和γ 氨基丁酸（GABA）神经递质系统和行为反应 这些系统都是很完善的。 关于这一点，我们所知甚少。 乙醇对氨基酸合成速率的影响 神经传递素 该项目的总体目标是开发 这些技术可以让我们测量乙醇的影响， 给药对神经递质合成速率的影响， 谷氨酸，GABA和天冬氨酸，从葡萄糖在完整的，清醒的大鼠。 神经递质合成速率和糖代谢模式 将使用13 C NMR光谱法来分析分布 同位素转化为葡萄糖代谢物。 这种技术允许一个 葡萄糖掺入代谢物的定量评价，包括 神经递质，通过确定“C”的分布， 代谢物中的碳原子。 长期目标是确定 乙醇通过糖酵解对葡萄糖代谢的影响， 回补和氧化途径，以便与葡萄糖 神经递质合成速率的变化。 研究 将使用一剂乙醇刺激运动， 活性（0.25 g/kg体重）和一个具有镇静作用的 微生物（1.0 g/kg体重）。 预计这两种剂量将 影响葡萄糖代谢和神经递质合成的模式 不同的是，因为以前已经证明，他们发挥 对某些大脑区域葡萄糖摄取的相反影响。 还将开发技术来研究乙醇对葡萄糖的影响 和神经递质代谢的混合物中分离的星形胶质细胞和 神经元（共培养物）。 一旦后一种技术建立起来，它将 在未来的研究中使用，以区分直接和间接 乙醇对大脑中葡萄糖和神经递质代谢的影响， 尤其是大脑皮层 1：3C NMR光谱也将是 用于开发跟踪葡萄糖的技术， 共培养物中的神经递质代谢。

APPLICANT'S ABSTRACT: The effects of ethanol on the glutamate and gamma amino butyric acid (GABA) neurotransmitter systems, and behavioral responses involving these systems, are well established. Less is known of the influence of ethanol on the rates of synthesis of the amino acid neurotransmitters. The overall Objective of this project is to develop techniques that will allow us to measure the effects of ethanol administration on the rates of synthesis of the neurotransmitters, glutamate, GABA and aspartate, from glucose in the intact, conscious rat. The rates of neurotransmitter synthesis and patterns of glucose metabolism will be determined using "C NMR spectroscopy to analyze for the distribution of isotope into glucose metabolites. This technique allows for a quantitative evaluation of glucose incorporation into metabolites, including the neurotransmitters, by identifying the distribution of "C into particular carbon atoms within a metabolite. A long range goal is to determine the impact of ethanol on the metabolism of glucose through the glycolytic, anaplerotic and oxidative pathways in order to relate alterations in glucose metabolism to changes in the rates of neurotransmitter synthesis. Studies will be carried out utilizing a dose of ethanol which stimulates motor activity (0.25 g/kg body weight) and one which has a -sedative effect on the organism (1.0 g/kg body weight). It is expected that these two doses will influence the patterns of glucose metabolism and neurotransmitter synthesis differently since it has been previously demonstrated that they exert opposite effects on the uptake of glucose in certain brain regions. Techniques will also be developed to study the effects of ethanol on glucose and neurotransmitter metabolism in mixtures of isolated astrocytes and neurons (co-cultures). Once this latter technology is established, it will be utilized in future studies to differentiate between direct and indirect effects of ethanol on glucose and neurotransmitter metabolism in the brain, particularly in the cerebral cortex. 1:3C NMR spectroscopy will also be utilized in development of the techniques for following glucose and neurotransmitter metabolism in the co-cultures.