The role of T-type calcium channels in the morphine effects in the nonspecific thalamus

项目摘要/摘要 人们对识别吗啡介导的效应和阿片类药物的复杂机制很感兴趣 以扩大新疗法的策略。尽管非特异性丘脑在调节中的作用 意识的作用已经有了很好的记录，它在吗啡介导的效应和成瘾中的可能作用并不是很好。 学习。首席研究员(PI)最近发现了T-型钙通道CaV3.1亚型的作用 丘脑中央内侧核(CEM)T通道在神经元兴奋性调节中的作用 麻醉期间丘脑皮质的震荡。此外，基于新的令人兴奋的初步数据， 该方案的主要目标是确定丘脑T通道CaV3.1亚型在脑内的功能作用。 吗啡的作用和成瘾。希望这能为新的治疗方法奠定基础。 和/或预防阿片类药物过度使用。在目标1中，我将使用膜片钳记录和双光子钙成像 从急性脑片体外和投射特异性靶向和操纵CEM神经元来确定 吗啡暴露的急性和持久影响(在反复暴露吗啡和吗啡诱导后 条件性位置偏爱-CPP)对CEM神经元T通道和兴奋性的影响。在《目标2》中，我将使用 与成瘾相关的行为学实验：CPP和自我给药(SA)模型，体内病毒沉默 技术(使用小发夹RNA(ShRNA)和Cre特异性基因敲除(KO))和化学发生方法 以探讨丘脑T通道在成瘾中的功能作用。 这些目标的完成将对吗啡效应和阿片类药物的机制产生新的见解 可能会发现新的治疗方案的毒瘾。PI将在以下方面接受指导和技术培训 上瘾行为和病毒技术，由电生理学和病毒技术专家在《动机》中 行为。科罗拉多大学和麻醉学系提供特殊的设施和 完成建议的实验所需的资源，以及拥有卓越声誉和记录的 在早期指导和过渡到独立调查人员方面的成就。建议数 培训、教育和研究将为PI提供技术和专业培训，使其成为一名 成功的独立调查员。

PROJECT SUMMARY / ABSTRACT There is substantial interest in identifying intricate mechanisms of the morphine-mediated effects and opioid addiction in order to expand strategies for novel therapies. Although the role of nonspecific thalamus in regulation of consciousness is well documented, its possible role in the morphine-mediated effects and addiction is not well studied. Principal Investigator (PI) has recently discovered the role of CaV3.1 isoform of T-type calcium channels (T-channels) in the central medial nucleus of thalamus (CeM) in neuronal excitability regulation and thalamocortical oscillations during anesthesia. Furthermore, based on the new exciting preliminary data, the overarching goal of this proposal is to determine the functional role of CaV3.1 isoform of thalamic T-channels in the morphine effects and addiction. It is hoped that this may form foundation for novel approaches to treatment and/or prevention of opioid overuse. In Aim 1, I will use patch-clamp recordings and 2-photon calcium imaging from acute brain slices ex vivo and projection-specific targeting and manipulating CeM neurons to determine acute and lasting effects of morphine exposure (after repeated morphine exposure and morphine-induced conditioned place preference – CPP) on T-channels and excitability of CeM neurons. In Aim 2, I will use behavioral experiments relevant to the addiction: CPP and self-administartion (SA) models, in vivo viral silencing technologies (with small hairpin RNA (shRNA) and Cre-specific knocking-out (KO)) and chemogenetic method for transient behavioral modulation to investigate functional role of thalamic T-channels in addiction. Completion of these aims will generate new insights into the mechanisms of the morphine effects and opioid addiction that could identify new treatment options. The PI will receive mentorship and technical training in addictive behavior and viral technologies by experts in electrophysiology and viral technologies in motivated behaviors. University of Colorado and the Department of Anesthesiology provide exceptional facilities and resources for completing the proposed experiments, as well as having an exceptional reputation and record of accomplishment for mentoring and transitioning early-stage into independent investigators. The proposed training, education and research will provide the PI with the technical and professional training to become a successful, independent investigator.