MOLECULAR MECHANISMS REGULATING NPY SYNTHESIS IN THE CNS

这是一个研究科学家发展奖二级的要求。 神经肽被认为是信息的组成部分 在中枢神经系统（CNS）中的转移。 他们是广泛的，但 离散，分布，并有助于一些生理和 电生理功能 分子生物学研究进展 允许克隆和测序的cDNA和基因编码 各种神经肽的前体蛋白。 这些研究 提供了神经肽基因表达，前体 蛋白质结构和神经肽表达的明显异常 几种中枢神经系统疾病 一种最近发现的肽， 神经肽Y（neuropeptide Y，NPY）是目前在中枢神经系统中发现的最丰富的肽 并支持诸如调节 心血管系统、摄食行为和皮质功能。 尽管 对于NPY的cDNA和基因的鉴定，知之甚少 关于神经肽Y生物合成和基因表达的调节， CNS。 这项研究将涉及神经肽Y合成的调控 通过“突触外”和“跨突触”机制。 “额外的- 将研究下丘脑NPY合成的“突触”调节 在设计用于研究分子机制的模型系统中， 调节摄食行为。 拟议的研究将确定在 葡萄糖和胰岛素对下丘脑神经肽Y的调节作用 正常动物和遗传性肥胖模型中的神经元（Zucker 肥鼠）。 研究NPY的“跨突触”调节 合成，将制备体外海马和下丘脑切片 以及各种药理学试剂的作用， 直接调节NPY表达或调节海马神经元 活动，将确定。 NPY表达的调节和 将通过核连续测定、核酸酶保护 分析前神经肽Y原mRNA含量、生物合成标记和 NPY的纯化和放射免疫分析。 拟议的研究将 联合收割机将定义明确模型系统与各种分子 生物技术，以独特地解决的几个方面， 调节NPY的表达和合成。 而且 拟议的研究将提供深入了解之间的相互作用， 外周和CNS调节神经肽的表达，因此， 理解机制多样性的根本重要性 调节行为和中枢神经系统功能。

This is a request for a Research Scientist Development Award Level II. Neuropeptides are recognized as integral components of information transfer in the central nervous system (CNS). They are widely, but discretely, distributed and subserve a number of physiological and electrophysiological functions. Recent progress in molecular biology has allowed the cloning and sequencing of the cDNAs and genes encoding the precursor proteins for a variety of neuropeptides. These studies have provided information on neuropeptide gene expression, precursor protein structure, and apparent abnormalities in neuropeptide expression in several CNS disorders. One recently identiifged peptide, neuropeptide Y (NPY), is the most abundant peptide yet found in the CNS and subserves such physiological functions as regulation of the cardiovascular system, feeding behavior and cortical function. Despite the identification of the cDNA and gene for NPY, little is known regarding the regulation of NPY biosynthesis and gene expression in the CNS. The proposed research will address the regulation of NPY synthesis by both "extra-synaptic" and "Transsynaptic" mechanisms. The "extra- synaptic" modulation of hypothalamic NPY synthesis will be investigated in model systems designed to investigate the molecular mechanisms regulating feeding behavior. The proposed studies will determine in vivo and in vitro how glucose and insulin modulate hypothalamic NPY neurons in normal animals and in a model of genetic obesity (the Zucker fatty rat). To investigate "trans-synaptic" regulation of NPY synthesis, in vitro hippocampal and hypothalamic slices will be prepared and the effects of various pharmacological agents, known to either directly modulate NPY expression or to modulate hippocampal neuronal activity, will be determined. Modulation of NPY expression and synthesis will be monitored by nuclear run-on assay, nuclease protection analysis of preproNPY mRNA content, biosynthetic labeling and purification of NPY and radioimmunoassay. The proposed studies will combine well-defined model systems with a variety of molecular biological techniques to uniquely address several aspects of the regulation of the expression and synthesis of NPY. Moreover, the proposed studies will provide insights into the interactions between the periphery and CNS to modulate neuropeptide expression and, thus, are of fundamental importance to understanding the diversity of mechanisms regulating behavior and CNS function.