EXCITATORY AMINO ACID REGULATION OF VASOPRESSIN CELLS

这项提议的主要目标是理解 谷氨酸受体的活性控制释放和合成 大细胞神经内分泌细胞(MNC)的加压素(VP)。 谷氨酸受体的几个亚型在VP上表达 神经内分泌细胞和大多数其他神经系统一样，发挥着 在控制兴奋性活动中的主要作用。药理作用 和寡核苷酸基因敲除方法将用于主要 培养神经内分泌细胞以确定NMDA的作用 受体亚基NR2B和NR2C与代谢性受体 MGluR1和mGluR3亚型在血管加压素和 加压素基因和多肽含量的调节。通过 比较释放与多肽和信使核糖核酸含量变化 在可控的刺激条件下，我们将开始 了解发布-合成过程中的这三个主要步骤 循环是协调的。渗透压、细胞体积控制的相互作用 谷氨酸受体的机制和外源性递质输入 激活将在体外和体内进行检测，以更好地了解 谷氨酸能活动是如何受这些刺激调节的。这个 每种受体亚型动员两个关键秒的能力 信使，钙和一种新的c-jun激酶，将在 使用数字视频成像和免疫组织化学进行详细说明。 这些实验将提供半定量的数据 在细胞水平上更清楚地描述受体的作用 确定天然谷氨酸受体的功能多样性 子类型。大细胞神经内分泌细胞是理想的 这样的亚型功能研究，因为他们有专门的， 定义功能和可量化的输出。VP的各种行为都有 与心血管调节、应激的控制有关 反应、热性惊厥、脑脊液产生、癫痫、学习和 记忆、社会依恋的形成、阿尔茨海默病和 缺氧缺血性脑损伤。《公约》揭示的基本原则 拟议的研究将提供有关 NMDA和代谢型谷氨酸受体亚型在中枢神经系统中的作用 跨越许多学科的正常和病理过程。

The major goal of this proposal is to understand how excitatory activity at glutamate receptors control the release and synthesis of vasopressin (VP) in magnocellular neuroendocrine cells (MNCs). Several subtypes of glutamate receptors are expressed on VP neuroendocrine cells and as in most other neuronal systems play a principle role in the control of excitatory activity. Pharmacological and oligonucleotide knockout approaches will be used on primary cultures of neuroendocrine cells to identify the role of NMDA receptor subunits NR2B and NR2C and metabotropic receptor subtypes mGluR1 and mGluR3 in the secretion of vasopressin and regulation of vasopressin mRNA and peptide content. By comparing release with changes in peptide and mRNA content under controlled conditions of stimulation, we will begin to understand how these three major steps in the release-synthesis cycle are coordinated. Interactions of osmotic, cell volume control mechanisms and extrinsic transmitter inputs with glutamate receptor activation will be examined in vitro and in vivo to better understand how glutamatergic activity is conditioned by these stimuli. The ability of each receptor subtype to mobilize two key second messengers, calcium and a novel c-jun kinase, will be examined in detail with digital video imaging and immunohistochemistry. These experiments will provide semi-quantitative data to characterize receptor actions at the cellular level and more clearly define the functional diversity of native glutamate receptor subtypes. The magnocellular neuroendocrine cells are ideal for such subtype-function studies since they have a dedicated, well define function and quantifiable output. Various actions of VP have been implicated in the control of cardiovascular regulation, stress responses, febrile seizures, CSF production, epilepsy, learning and memory, social attachment formations, Alzheimer's disease, and hypoxic ischemic brain damage. Basic principles revealed by the proposed studies will provide essential information about the function of NMDA and metabotropic glutamate receptor subtypes in normal and pathological processes across many disciplines.