REGULATION OF PREPRO-ORPHANIN FQ (OFQ)-DERIVED PEPTIDES

前孤啡肽原 FQ (OFQ) 衍生肽的调控

• 批准号: 2898178
• 负责人: RICHARD G ALLEN
• 金额: $ 17.11万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2898178
• 关键词: G protein; complementary DNA; genetically modified animals; glucocorticoids; hypothalamus; laboratory mouse; laboratory rat; ligands; neurons; neuropeptides; northern blottings; opioid receptor; polymerase chain reaction; protein metabolism; radioimmunoassay; receptor coupling; stimulant /agonist; tissue /cell culture

Recently, the endogenous peptide ligand for an opioid-like g protein-coupled receptor (called LC 132 or ORL-10 has been identified and named orphanin FQ [OFQ] or nociceptin. The cDNA has been cloned and initial observations suggest that OFQ is contained in a larger precursor molecule as ae the enkephalins, endorphins and dynorphins. There appears to be several other putative peptides contained in the OFQ precursor, all bounded by paired-basic amino acids, a motif shared by many prohomones. We hypothesize that several other biologically active peptides are derived from the OFQ precursor and that the release of these peptides, including OFQ are regulated by dopamine and morphine. In order to test these hypotheses we will: 1) Determine the processing of prepro(PP)- OFQ-derived peptides in the rodent hypothalamus. These studies will characterize pp-OFQ-derived peptide liberated from the precusor and provide evidence that the peptides other than OFQ that are predicted from the cDNA are produced in hypothalamic neurons. 2) characterize the regulation of OFQ in monolayer cultures of hypothalamic neurons. Monolayer cultures of rat hypothalamic neurons will be incubated in the presence of ppiate receptor agonists or dopamine and the culture medium assayed for OFQ immunoreactivity. We will also continue to develop radioimmunoassays (RIAs) for each of the other neuropeptides potentially derived from pp-OfQ and use these assays to monitor the regulation of their secretion by dopamine and drugs of abuse. 3) Determien of glucocorticoids regulate pp-OFQ mRNA and peptide levels in vivo and ni vitro. We hypothesisze that OFQ-derived peptides are majormodulators of the stress response. We will use transgenic mie that have negligible circulating glucosteroids and subject them to acute and chronic restraint-stress. Hypothalamic OFQ peptide levlels will be measured by RIA and OFQ mRNA levels using molecular billogical methods. These studies will reveal whether OFQ levels are regulated by glucosteroids in the intact animal and thus potentially responsive to stress paradigms.

最近，用于阿片类药物的内源性肽配体 蛋白质偶联受体（称为LC 132或ORL-10已是 识别并命名为孤儿FQ [OFQ]或NociTptin。 这 cDNA已被克隆，最初的观察结果表明 OFQ作为AE较大的前体分子中包含 Enkephalins，内啡肽和Dynorphins。 似乎有 OFQ前体中包含的其他几种推定肽， 全部由配对基础氨基酸界定，这是一个由 许多动物。 我们假设其他几个 生物活性肽是从OFQ前体得出的 并且这些肽的释放，包括OFQ 由多巴胺和吗啡调节。 为了测试这些 假设我们将：1）确定Prepro（PP）的处理 - 啮齿动物下丘脑中的OFQ衍生肽。 这些 研究将表征从PP-OFQ衍生的肽中解放的 precustor并提供证据表明除 从cDNA预测的OFQ是在 下丘脑神经元。 2）表征OFQ的调节 下丘脑神经元的单层培养物。 单层 大鼠下丘脑神经元的培养物将在 Ppiate受体激动剂或多巴胺的存在以及 培养基分析了OFQ免疫反应性。 我们将 也继续为每一个开发放射免疫测定（RIA） 可能衍生自PP-OFQ的其他神经肽和 使用这些测定法监测其分泌的调节 多巴胺和滥用药物。 3）确定糖皮质激素 调节体内和Ni体外的PP-OFQ mRNA和肽水平。 我们假设OfQ衍生的肽是 压力反应的Majormodulator。 我们将使用转基因 MIE可忽略不计的循环糖尿类固醇和受试者 他们急性和慢性约束压力。 下丘脑OFQ 肽左带将通过RIA和OFQ mRNA水平测量 使用分子收入方法。 这些研究将揭示 OFQ水平是否由完整的葡萄糖类固醇调节 动物，因此有可能响应压力范例。