NEURONAL NICOTINIC RECEPTORS FUNCTION, PHARMACOLOGY AND MODULATION BY RNA APTAME

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 神经型烟碱型乙酰胆碱受体(NAChR)被认为是 中枢(CNS)和外周(PNS)神经系统的神经传递。少校 在CNS和PNS中发现的nAChRs有α-7、α-4/β-2和α-3/β-4亚型。 NAChR的激活剂(激动剂)、抑制剂(拮抗剂)和调节剂已被用作 用于治疗阿尔茨海默氏症、帕金森氏病和其他疾病的药物 神经精神疾病，如精神分裂症、注意力缺陷、情绪和焦虑 精神障碍和慢性疼痛。我们有兴趣研究4S，6S- 西伯拉烯二醇(4S)，一种在烟草中发现的丰富的西门冬醇类化合物 α3β4 nAChRs。以前的工作表明，烟草西比林类化合物在 大脑中的大量尼古丁可以改变尼古丁的作用。这个项目的目的是 开发可抑制和/或标记α3β4尼古丁的受体结合RNA适配子 4S膜状部位的受体。SELEX技术产生的RNA适配子 (用指数富集法进行配体的系统进化)具有高亲和性 与包括膜结合受体在内的复杂蛋白质分子结合的亲和配体。 在这个项目中，我们将使用SELEX选择使用膜的RNA适配子 从稳定表达α3β4受体的细胞系中制备的蛋白质 4S西膜类取代受体结合的适配子。最后一批适配子将是 根据其nAChR亚型特异性和对 受体功能。亚型选择性适配子将被荧光素标记并用于 描述神经网络中nAChR的相互作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Neuronal nicotinic acetylcholine receptors (nAChR) are recognized as important elements in neurotransmission in the central (CNS) and peripheral (PNS) nervous systems. The major nAChRs found in the CNS and PNS are alpha-7, alpha-4/beta-2, and alpha-3/beta-4 subtypes. Activators (agonists), inhibitors (antagonists), and modulators of nAChR have been used as drug leads for treatment in Alzheimer's disease Parkinson's disease, and other neuropsychiatric illnesses such as schizophrenia, attention deficit, mood, and anxiety disorders, and chronic pain. We are interested in studying the effects of 4S, 6S- cembratrienediol (4S), an abundant cembranoid compound found in tobbaco on the alpha3beta4 nAChRs. Previous work indicate that tobbaco cembranoids accumulate in the brain in significant amounts to modify the action of nicotine. The purpose of this project is to develop receptor-binding RNA aptamers that can inhibit and/or label alpha3beta4 nicotinic receptors at the 4S cembranoid sites. RNA aptamers generated by the SELEX technique (Systematic Evolution of Ligands by Exponential enrichment) have the properties of high- affinity ligands binding to complex protein molecules including membrane-bound receptors. For this project, we will employ the SELEX selection of RNA aptamers using membrane proteins prepared from cell lines stably expressing alpha3beta4 receptors in the presence of 4S cembranoids to displace the receptor-bound aptamers. The final pool of aptamers will be sequenced and characterized according to their nAChR subtype specificity and effects on receptor function. Subtype-selective aptamers will be fluorescein-labeled and used to characterize nAChR interplay in neuronal networks.