MRNA & PEPTIDE SYNTHESIS OF VASOPRESSIN AND OXYTOCIN

拟议的实验旨在检测，在 单个细胞，生理学上的变化 对加压素能和催产素能有意义的刺激 神经元。人们对这些神经元了解很多，但之前 调查研究了信使核糖核酸水平和蛋白水平的全球变化 求婚。表面上均一的单个神经元 解剖核团的传入和传出不同 连接性，以及在多肽共定位方面。因此，在 应采用神经内分泌功能的方法 如果激素反应仅限于特定的加压素，则将其分离 或者催产素细胞。利用定量原位杂交 测定加压素和催产素的相对含量 水平，我将检查是否所有激素合成细胞 在下丘脑细胞群内对某些生理反应 挑战，或者特定的细胞亚群是否调节激素 回应。基于对细胞连通性的研究，以及 下面描述的其他因素，假设荷尔蒙 反应由特定的加压素能和催产素能介导 细胞。原位杂交与免疫细胞化学相结合 对于多肽共定位和轨迹跟踪技术，我 希望确定动态的、解剖学的反应模式 这些神经系统的活体，以及阐明 这些细胞的显著特征就是这些细胞 对动态平衡挑战的反应。 除了提供了一个模型，说明如何使用单个 神经元是在神经内分泌的背景下整合的 系统，这些研究可能具有临床意义。第一，疾病 扰乱合成、运输或调节的过程 释放加压素可引起不适当的代偿 体液需求。拟议中的涉及水的实验 平衡和神经垂体系统的反应是相关的。 其次，加压素在高血压和应激中起作用， 对单细胞水平的神经元反应的研究将提供 有关自主或自主的差异反应的信息 神经血液介导的加压素能神经元与 神经垂体神经元。最后，催产素基因表达是一种 可能与临床相关的几乎未被探索的物质 催产素在怀孕、哺乳和压力中的作用。

The proposed experiments are designed to detect, at the level of the single cell, the alterations that follow physiologically meaningful stimulation of vasopressinergic and oxytocinergic neurons. Much is known about these neurons, but previous investigations studied global alterations in levels of mRNA and proposes. Individual neurons within apparently homogeneous anatomical nuclei differ in terms of their afferent and efferent connectivity, and in terms of peptide co-localization. Hence, in approach to neuroendocrine function should be employed tbat can detact if hormone response is restricted to specific vasopressin or oxytocin cells. Utilizing quantitative in situ hybridization to determine relative amounts of vasopressin and oxytocin mRNA levels, I will examine whether all hormone-synthesizing cells within a hypothalamic cell group response to certain physiological challenges, or whether specific subgroups of cells mediate hormone response. Based upon investigations of cell connectivity, and other factors described below, it is hypothesized that hormone response is mediated by specific vasopressinergic and oxytocinergic cells. Combining in situ hybridization with immunocytochemistry for peptide co-localization, and with tract-tracing techniques, I hope to determine the dynamic, anatomical pattern of response in vivo, of these neural systems, and the basis for elucidating the salient characteristics of those cells that specifically mediate response to homeostatic challenge. In addition to providing a model for how the function of single neurons is integrated within the context of a neuroendocrine system, these studies may have clinical relevance. First, disease processes that disrupt the regulation of synthesis, transport, or release of vasopressin can precipitate inappropriate compensation of bodily fluid needs. Proposed experiments that involve water balance and how the neurohypophysial system responds are relevant. Second, vasopressin plays a role in hypertension and stress, and study of neuronal response at the single cell level will provide information regarding differential response in autonomic or neurohaemal-mediating vasopressinergic neurons compared to neurohypophysial neurons. Finally, oxytocin gene expression is an almost unexplored matter that may be clinically relevant to oxytocin's role in pregnancy, lactation, and stress.