Photoactivatable ligands for nicotinic optopharmacology

项目总结 长期接触烟草产品中的尼古丁会导致许多健康后果(肺癌， 肺气肿、高血压等)每年造成600多万人死亡。复发率很高，在 那些试图戒烟的人和寻求促进戒烟的药物疗法 效果适中。因此，对治疗尼古丁的更有效策略的需求还远远没有得到满足。 依赖。发展这样的策略需要对生物学的更详细的理解 导致尼古丁成瘾的机制。与nAChRs机制研究相关的一个基本目标是 更好地了解nAChRs在单个神经内不同部位的位置和活性 细胞。尽管一些基础研究已经开始描述nAChR的亚细胞分布，但有 目前还没有可行的方法在亚细胞水平上对nAChRs进行功能性询问。这意味着我们是 目前无法确定尼古丁介导的nAChRs的重要功能改变 见于树突、轴突、突触前终末或神经元胞体。回答这个关键问题是 对于该领域充分了解尼古丁依赖的分子和细胞基础来说，这是绝对必要的。 在这里，我们提出了一个R21/R33阶段性创新项目，直接解决我们能力中的这些关键差距 研究天然的nAChRs。R21阶段将形成并表征一种光活化尼古丁(PA-NIC) 用于尼古丁“去除”实验的化合物。R21目标1专注于识别1个或多个 使用紫外光/可见光光学方法的合适化合物，而目标2将表征有希望的 使用双光子去离子化技术的化合物，提供了增强的空间分辨率。在R33阶段，我们 建议在发现实验中使用这些创新的化合物和光学方法，旨在 了解有关nAChR函数的新细节。R33目标1将涉及关键细胞上nAChRs的功能映射 涉及尼古丁依赖的类型。R33 Aim 2将探索这些受体如何使尼古丁参与 在大脑奖赏系统中神经传递的电路级调制中。这个项目代表着一个重要的 胆碱能生物领域的技术进步，因为它不仅将产生广泛使用的新工具，而且 将利用这些工具来揭示有关尼古丁依赖的新的机械细节。

PROJECT SUMMARY Chronic exposure to nicotine in tobacco products results in numerous health consequences (lung cancer, emphysema, hypertension, etc.) and accounts for over 6 million deaths per year. Relapse rates are high among those who attempt to quit smoking, and pharmacotherapies that seek to foster smoking cessation have moderate effectiveness. Thus, there is a significant unmet need for more effective strategies to treat nicotine dependence. Development of such strategies requires a more detailed understanding of the biological mechanisms leading to nicotine addiction. An essential goal related to mechanistic studies on nAChRs is gaining a better understanding of the location and activity of nAChRs in discrete sites within individual nerve cells. Although some basic research studies have begun to describe nAChR subcellular distribution, there is currently no plausible way to functionally interrogate nAChRs at the subcellular level. This means we are currently unable to determine whether the important nicotine-mediated functional alterations in nAChRs occur in dendrites, axons, presynaptic terminals, or in neuronal somata. Answering this key question is absolutely required for the field to fully understand the molecular and cellular basis for nicotine dependence. Here, we propose a R21/R33 phased innovation project that directly addresses these critical gaps in our ability to study native nAChRs. The R21 phase will develop and characterize a photoactivatable nicotine (PA-Nic) compound for use in nicotine “uncaging” experiments. R21 Aim 1 focusses on identification of 1 or more suitable compounds using ultraviolet/visible light optical methods, while Aim 2 will characterize promising compounds using 2-photon uncaging techniques, which offer enhanced spatial resolution. In the R33 phase, we propose to employ these innovative compounds and optical methods in discovery experiments designed to uncover new details about nAChR function. R33 Aim 1 will involve functional mapping of nAChRs on key cell types involved in nicotine dependence. R33 Aim 2 will probe how these receptors enable nicotine to participate in circuit-level modulation of neurotransmission in brain's reward system. This project represents a substantial technical advance for the cholinergic biology field, as it will not only produce new tools for widespread use, but will utilize those tools to uncover new mechanistic details about nicotine dependence.