MOLECULAR MECHANISMS REGULATING NPY SYNTHESIS IN THE CNS

中枢神经系统中调节 NPY 合成的分子机制

• 批准号: 3070174
• 负责人: JEFFREY D WHITE
• 金额: $ 5.36万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3070174
• 关键词: bombesin; central nervous system; complementary DNA; developmental genetics; disease /disorder model; endorphins; enkephalins; gene expression; genetic regulation; glucose; hypothalamus; in situ hybridization; insulin; laboratory rat; messenger RNA; molecular cloning; molecular pathology; neural information processing; neuropeptide Y; neurotransmitter biosynthesis; neurotransmitter metabolism; peptide chemical synthesis; protein biosynthesis; reflex; tissue /cell culture

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.

这是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）中转移。 他们很广泛，但是 离散，分布并提供许多生理和 电生理功能。 分子生物学的最新进展 允许cDNA和基因编码的克隆和测序 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 在几种中枢神经系统疾病中。 一个最近鉴定的肽， 神经肽Y（NPY）是CNS中最丰富的肽 以及这样的生理功能，例如调节 心血管系统，进食行为和皮质功能。 尽管 鉴定NPY的cDNA和基因，鲜为人知 关于NPY生物合成和基因表达的调节 CNS。 拟议的研究将解决NPY合成的调节 通过“突触外”和“透射性”机制。 “超级 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 调节喂养行为。 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 正常动物和遗传肥胖模型中的神经元（扎克 脂肪大鼠）。 调查NPY的“反突触”调节 合成，将制备体外海马和下丘脑切片 以及各种药理学剂的影响，已知 直接调节NPY表达或调节海马神经元 活动将被确定。 NPY表达和 合成将通过核跑步测定，核酸酶保护监测 分析预合体mRNA含量，生物合成标记和 NPY和放射免疫测定法的纯化。 拟议的研究将 将定义明确的模型系统与各种分子结合在一起 生物学技术以唯一解决的几个方面 调节NPY的表达和合成。 而且， 拟议的研究将为您提供有关互动的见解 外围和中枢神经系统调节神经肽表达，因此 对于理解机制多样性的根本重要性 调节行为和CNS功能。