EXCITATORY AMINO ACID REGULATION OF VASOPRESSIN CELLS

这项提议的主要目的是了解 谷氨酸受体的活性控制谷氨酸的释放和合成 大细胞神经内分泌细胞（MNCs）中的血管加压素（VP）。 几种谷氨酸受体亚型在VP上表达 神经内分泌细胞和大多数其他神经系统一样， 控制兴奋性活动的主要作用。 药理 和寡核苷酸敲除方法将用于原发性 培养神经内分泌细胞，以确定NMDA的作用 受体亚单位NR 2B和NR 2C和代谢型受体 mGluR 1和mGluR 3亚型在加压素分泌中的作用， 调节血管加压素mRNA和肽含量。 通过 比较释放与肽和mRNA含量的变化 在受控的刺激条件下，我们将开始 理解释放合成中的这三个主要步骤 周期是协调的。 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查，以更好地了解 这些刺激是如何制约脑电活动的 的 每个受体亚型动员两个关键的第二阶段的能力， 信使，钙和一种新的c-jun激酶，将在 详细的数字视频成像和免疫组织化学。 这些实验将提供半定量数据， 在细胞水平上表征受体作用， 确定天然谷氨酸受体的功能多样性 亚型 大细胞神经内分泌细胞是理想的 因为他们有一个专门的， 定义功能和可量化的输出。 VP的各种行动 与心血管调节、压力控制有关 反应、热性惊厥、CSF产生、癫痫、学习和 记忆，社会依恋形成，阿尔茨海默病， 缺氧缺血性脑损伤 基本原则揭示了 拟议的研究将提供有关 NMDA和代谢型谷氨酸受体亚型在 正常和病理的过程。

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.