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

T型钙通道在非特异性丘脑吗啡作用中的作用

• 批准号: 10525691
• 负责人: Tamara Timic Stamenic
• 金额: $ 16.91万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10525691
• 关键词: Acute; Addictive Behavior; Amygdaloid structure; Anesthesia procedures; Anesthesiology; Animal Behavior; Animal Model; Anterior; Behavior; Behavioral; Behavioral Model; Brain; Brain region; Calcium; Calcium Channel; Calcium ion; Cell Nucleus; Cells; Central Medial Thalamic Nucleus; Claustral structure; Closure by clamp; Colorado; Conscious; Corpus striatum structure; Coupled; Data; Diffuse; Disease; Drug Addiction; Drug Targeting; Drug abuse; Electrophysiology (science); FOS Protein; FOS gene; Foundations; Goals; Image; Injections; Intralaminar Nuclear Group; Knock-out; Knowledge; Laser Scanning Microscopy; Mediating; Mentors; Mentorship; Methods; Mind; Modeling; Molecular; Morphine; Neurons; Nucleus Accumbens; Olfactory tubercle; Opiate Addiction; Opioid; Opioid Analgesics; Overdose; Pain; Persons; Pharmaceutical Preparations; Pharmacology; Play; Positioning Attribute; Preparation; Prevention; Principal Investigator; Process; Property; Protein Isoforms; Regulation; Research; Research Personnel; Research Proposals; Resources; Role; Self Administration; Slice; Stimulus; T-Type Calcium Channels; Technology; Testing; Thalamic structure; Tissues; Training; Training and Education; United States Dept. of Health and Human Services; United States National Institutes of Health; Universities; Viral; addiction; base; biophysical properties; channel blockers; conditioned place preference; drug reward; education research; experimental study; genetic approach; in vivo; insight; interest; knock-down; motivated behavior; mouse genetics; mu opioid receptors; neuronal excitability; new therapeutic target; novel; novel strategies; novel therapeutics; opioid overuse; opioid use disorder; overdose death; patch clamp; putamen; skills; small hairpin RNA; two-photon; voltage

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.

项目概要/摘要 人们对确定吗啡介导作用和阿片类药物的复杂机制非常感兴趣 成瘾，以扩大新疗法的策略。尽管非特异性丘脑在调节中的作用 意识的作用已被充分记录，但其在吗啡介导的作用和成瘾中可能发挥的作用尚不清楚 研究过。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) 中的 T 通道在神经元兴奋性调节中的作用 麻醉期间的丘脑皮质振荡。此外，根据令人兴奋的新初步数据， 该提案的总体目标是确定丘脑 T 通道 CaV3.1 亚型的功能作用 吗啡效应和成瘾。希望这可以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 2 光子钙成像 从离体急性脑切片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（反复吗啡暴露和吗啡引起的 条件性位置偏好（CPP）对 T 通道和 CeM 神经元兴奋性的影响。在目标 2 中，我将使用 与成瘾相关的行为实验：CPP 和自我给药 (SA) 模型、体内病毒沉默 技术（小发夹RNA（shRNA）和Cre特异性敲除（KO））和化学遗传学方法 用于瞬时行为调节，以研究丘脑 T 通道在成瘾中的功能作用。 这些目标的完成将为吗啡效应和阿片类药物的机制产生新的见解 成瘾可以找到新的治疗选择。 PI 将接受以下方面的指导和技术培训： 电生理学和病毒技术专家的成瘾行为和病毒技术 行为。科罗拉多大学和麻醉科提供卓越的设施和 完成拟议实验的资源，以及拥有卓越的声誉和记录 在指导和过渡到早期阶段的独立调查员方面取得的成就。拟议的 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。