MOLECULAR MECHANISMS REGULATING NPY SYNTHESIS IN THE CNS

这是对研究科学家发展奖二级的请求。 神经肽被认为是信息的组成部分 中枢神经系统（CNS）的转移。 它们分布广泛，但是 离散地、分布地并促进许多生理和 电生理功能。 分子生物学最新进展 已允许对 cDNA 和编码基因进行克隆和测序 多种神经肽的前体蛋白。 这些研究 提供了有关神经肽基因表达、前体的信息 蛋白质结构和神经肽表达的明显异常 在多种中枢神经系统疾病中。 最近发现的一种肽， 神经肽 Y (NPY) 是中枢神经系统中迄今为止发现的最丰富的肽 并促进诸如调节等生理功能 心血管系统、进食行为和皮质功能。 尽管 NPY cDNA 和基因的鉴定，知之甚少 关于NPY生物合成和基因表达的调控 中枢神经系统。 拟议的研究将解决 NPY 合成的调控问题 通过“突触外”和“跨突触”机制。 那个“额外的 将研究下丘脑 NPY 合成的突触调节 在旨在研究分子机制的模型系统中 调节进食行为。 拟议的研究将确定 体内和体外葡萄糖和胰岛素如何调节下丘脑 NPY 正常动物和遗传性肥胖模型中的神经元（Zucker 脂肪鼠）。 研究 NPY 的“跨突触”调节 合成，将制备体外海马和下丘脑切片 以及已知的各种药物的作用 直接调节 NPY 表达或调节海马神经元 活动，将被确定。 NPY表达的调节和 合成将通过核连续分析、核酸酶保护进行监测 preproNPY mRNA 含量、生物合成标记和分析 NPY 的纯化和放射免疫分析。 拟议的研究将 将明确的模型系统与各种分子相结合 生物技术独特地解决了几个方面 NPY 表达和合成的调节。 此外， 拟议的研究将深入了解两者之间的相互作用 外周和中枢神经系统调节神经肽的表达，因此， 了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.