NEURONAL NICOTINIC RECEPTORS FUNCTION, PHARMACOLOGY AND MODULATION BY RNA APTAME

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 神经元烟碱乙酰胆碱受体（nAChR）被认为是神经系统中的重要元件。 中枢（CNS）和外周（PNS）神经系统中的神经传递。主要 在CNS和PNS中发现的nAChR是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 治疗阿尔茨海默病、帕金森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 疾病和慢性疼痛。我们感兴趣的是研究4S，6S- cembratrieneline（4S），一种在热带雨林的巴科中发现的丰富的西松烷类化合物， α 3 β 4 nAChRs。以前的工作表明，西松烷类化合物积累在 大脑中的大量改变尼古丁的作用。该项目的目的是 开发可抑制和/或标记α 3 β 4烟碱的受体结合RNA适体 4S西松烷酸受体。通过SELEX技术产生的RNA适体 （通过指数富集的配体的系统进化）具有高- 与包括膜结合受体的复杂蛋白质分子结合的亲和配体。 在这个项目中，我们将使用膜选择RNA适体的SELEX选择， 由在存在下稳定表达α 3 β 4受体的细胞系制备的蛋白质 4S西松烷类化合物置换受体结合的适体。适体的最终池将是 根据它们的nAChR亚型特异性和对 受体功能亚型选择性适体将被荧光素标记并用于 表征nAChR在神经元网络中的相互作用。

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.