AFFERENT CONTROL OF NEUROPEPTIDE SYNTHESIS

这个项目的具体目的是提供时间和机会 让PI学习如何利用急性切片和器官型切片 生理调节背景下的培养准备 神经肽基因表达。联营公司的长期目标 研究项目是确定传入信号传递机制， 边缘前脑神经的细胞反应和地形图 帮助调节老鼠体内动态平衡的回路。这些电路是 对调节行为、自主神经和内分泌反应至关重要 动物对动态平衡紊乱的反应。中心假设是 动态平衡紊乱会修改化学编码的信息 在神经元内，通过调制编码单个和联合- 表达的神经肽。这些特定于细胞和刺激的修饰 促进动物的适当反应，要么通过调节 自主神经系统的活动，神经内分泌功能， 或者可能通过修改中央图案生成器来开发和 规范目标导向的行为。因为相对简单， 潜在的生理和行为，以及广泛的 文献，整个项目集中在调查 控制流体平衡的回路和机构的组织 那只老鼠。在此背景下，本项目的目标是学习如何 使用体外技术来研究潜在的细胞 下丘脑多肽基因表达的调控机制 室旁核(PVH)和杏仁中央核(CEA) 在大鼠体内体液平衡发生改变后。将这些结果联系起来 重要且有充分记录的模型允许对数据进行解释 在令人信服的背景框架内，这是许多其他项目无法实现的 目前使用的是“压力”模型。所使用的化验方法-主要是 原位杂交和免疫细胞化学-将允许检测 PVH和CEA中CRH、前脑啡肽和神经降压素mRNAs的变化 在糖皮质激素或递质操纵后。用这些体外培养的 技术，私家侦探将学习使用传统药理学和 新近介绍的反义寡核苷酸阻断血管内皮生长因子的方法 为此目的受体翻译。另一个目标是看看其他人是如何 体外技术(如电生理学、分子技术)可能是 适应了国际和平研究所的研究项目。协作、协商和 与PI所在机构和其他地方的专家教师进行互动 将提供专业知识来完成 程序。设施将由该协会的机构提供。在漫长的岁月里 术语，研究地形和组织的机制 流体平衡的动态平衡方面将为 了解相对简单的哺乳动物的神经组织 在细胞、系统和行为层面上的行为。会的 最终使我们能够解决许多临床疾病(例如。 高血压、肥胖症、饮食失调)目前至关重要 对人类健康来说，这在核心上扰乱了动态平衡的调节 他们的病因。

The specific aim of this project is to provide the time and opportunity for the PI to learn how to utilize acute slice and organotypic slice culture preparations within the context of physiological regulation of neuropeptide gene expression. The long term goal of the associated research project is to identify the afferent signalling mechanisms, cellular responses, and topography of the limbic forebrain neural circuits that help regulate homeostasis in the rat. These circuits are critical for regulating the behavioral, autonomic and endocrine response of the animal to homeostatic disturbance. The central hypothesis is that homeostatic disturbances modify chemically-coded information contained within neurons by modulating the mRNAs that code for singly- and co- expressed neuropeptides. These cell- and stimulus-specific modifications facilitate the appropriate response by tile animal, either by modulating the activity of the autonomic nervous system, neuroendocrine function, or perhaps by modifying the central pattern generators that develop and regulate goal-directed behaviors. Because of the relative simplicity of the underlying physiology and behavior, along with an extensive literature, the overall project concentrates on investigating the organization of the circuits and mechanisms controlling fluid balance in the rat. Within this context, the goals of this project are to learn how to use in vitro techniques to investigate the underlying cellular mechanisms which modulate peptide gene expression in the hypothalamic paraventricular nucleus (PVH) and central nucleus of the amygdala (CEA) after alterations in fluid homeostasis in the rat. Relating these results to important and well documented models allows the interpretation of data within a compelling and contextual framework not possible with many other currently used 'stress' models. The assay methods used-principally in situ hybridization and immunocytochemistry-will allow the detection of changes in CRH, proenkephalin and neurotensin mRNAs in the PVH and CEA after glucocorticoid or transmitter manipulation. With these in vitro techniques, the PI will learn to use conventional pharmacology and recently introduced methods of antisense oligonucleotide blockade of receptor translation to this end. A further goal is to see how other in vitro techniques (eg. electrophysiology, molecular techniques) might be adapted to the PI's research project. Collaborations, consultations, and interactions with expert faculty in the PI's institution, and elsewhere in Southern California will provide the expertise to complete the program. Facilities will be provided by the PI's institution. In the long term, investigating the topography and mechanisms that organize the homeostatic aspects of fluid balance will provide a working model for understanding the neural organization of a relatively simple mammalian behavior at the cellular, systems and behavioral level. It will eventually allow us to address many of the clinical disorders (eg. hypertension, obesity, eating disorders) currently of central importance to human health, that have perturbed homeostatic regulation at the core of their etiology.