AFFERENT CONTROL OF NEUROPEPTIDE SYNTHESIS

该项目的具体目标是提供时间和机会， 让PI学习如何使用急性切片和器官型切片 在生理调节的背景下， 神经肽基因表达联合国的长期目标 研究项目是确定传入信号机制， 细胞反应和边缘前脑神经元的地形图 帮助调节大鼠体内稳态的回路。这些电路 对于调节行为、自主神经和内分泌反应至关重要 动物体内平衡紊乱。核心假设是， 稳态紊乱改变了化学编码信息 在神经元内，通过调节编码单-和共- 表达的神经肽。这些细胞和刺激特异性的修饰 促进动物的适当反应，或者通过调节 自主神经系统的活动，神经内分泌功能， 或者也许通过修改中央模式发生器， 调节目标导向的行为。由于相对简单， 潜在的生理和行为，沿着广泛的 文学，整个项目集中在调查 控制流体平衡的回路和机构的组织 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 调节下丘脑中肽基因表达的机制 室旁核（PVH）和杏仁中央核（CEA） 在大鼠体液平衡改变后。将这些结果与 重要的和有据可查的模型允许解释数据 在一个令人信服的背景框架内， 目前使用的“压力”模型。使用的测定方法-主要用于 原位杂交和免疫细胞化学将允许检测 PVH和CEA中CRH、前脑啡肽原和神经降压素mRNA的变化 在糖皮质激素或递质操纵之后。用这些体外培养的 技术，PI将学习使用传统药理学， 最近引入的反义寡核苷酸阻断 转基因为此另一个目标是看看其他在 体外技术（例如，电生理学，分子技术）可能是 适应PI的研究项目。合作、协商和 与PI机构和其他地方的专家教师互动 在南加州将提供专业知识， 程序.设施将由PI所在机构提供。从长远 术语，调查组织的地形和机制， 体液平衡的稳态方面将提供一个工作模型， 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。它将 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）， 对人类健康的影响， 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.