Determining how neurotensin mediates valence processing and compulsive cocaine seeking

项目概要/摘要 物质使用障碍的一个标志是尽管有严重的不良后果，但仍继续使用药物。而 对啮齿动物的研究表明，利用积极和消极的效价处理可以影响 这种寻求奖励的强迫性行为模式与惩罚相冲突， 调节效价加工和指导最终行为选择的机制仍然未知。研究 结果表明，基底外侧杏仁核（BLA）优先投射到丘脑外侧核（BLA-NAc）， 编码阳性效价，包括蔗糖和可卡因预测线索和驱动器接近行为，而 向杏仁核中央核（BLA-CeM）的投射编码负价和驱动回避。 这一证据有力地表明，BLA-NAc和BLA-CeM神经元可以编码价信息加工 在强迫性可卡因寻求过程中，并在持续的行为过程中集体调解决策。这 建议的项目旨在确定指导BLA-NAc之间的价分配的基本神经底物 和BLA-CeM神经元（K99期），并检查是否相同的机制有助于编码 可卡因成瘾（R 00阶段）初步数据表明，神经降压素（NT），一个13个氨基酸 肽，在介导BLA中的效价加工中起关键作用。特别是，激活的终端 丘脑室旁核（PVT）NT神经元在BLA增强奖赏学习和损害 惩罚学习，而PVT-BLA NTergic信号的中断产生相反的行为 方面的影响.在K99阶段，申请人将接受CRISPR-Cas9介导的培训。 不影响谷氨酸和应用机器的基因操作来询问NTergic贡献 基于学习的计算机视觉工具和分类算法，以提取不同的行为图案， 解码神经关联这项培训将使申请人调查的具体作用NT价 BLA神经元在效价加工过程中的分配和编码特性。为了验证这一点，申请人将 检查CRISPR介导的Nt基因敲低对BLA投射PVT神经元的影响， 奖励和惩罚学习（目标1）以及击倒对价编码特性的影响。 BLA-NAc和BLA-CeM神经元在奖励和惩罚辨别任务（目的2）。在确保 作为一个独立的研究职位，申请人将通过结合这些职位来开始该奖项的R 00阶段 从K99阶段收集的方法与申请人的专业知识，在研究可卡因成瘾与自我， 管理和惩罚奖励寻求范式，调查如何PVT NTergic输入调节 强迫性可卡因寻求的编码（目标3）。在一起，训练和科学的进步， 通过这个项目的完成实现将允许申请人获得终身教职 在一所顶尖的研究型大学，并提供必要的关键试点数据，以准备第一个R 01申请研究 更多的分布式神经通路控制着动机性行为和强迫性药物寻求。

PROJECT SUMMARY/ABSTRACT A hallmark of substance use disorders is continued drug use despite profound adverse consequences. While studies in rodents have suggested that leveraging both positive and negative valence processing can affect this compulsive pattern of reward-seeking behavior in conflict with punishment, the underlying mechanisms that regulate valence processing and guide ultimate behavioral selections remain unknown. Studies have shown that the basolateral amygdala (BLA) projections to the nucleus accumbens (BLA-NAc) preferentially encode positive valence, including sucrose and cocaine predictive cues and drive approach behaviors, while projections to the central nucleus of the amygdala (BLA-CeM) encode negative valence and drive avoidance. This evidence strongly suggests that BLA-NAc and BLA-CeM neurons could encode valence processing during compulsive cocaine seeking and collectively mediate decision-making during ongoing behavior. This proposed project aims to identify basic neural substrates that guide the valence assignment among BLA-NAc and BLA-CeM neurons (K99 Phase), and examine whether the same mechanism contributes to the encoding of compulsive cocaine-seeking (R00 Phase). Preliminary data suggest that neurotensin (NT), a 13 amino acid peptide, plays a critical role in mediating valence processing in the BLA. Specifically, activation of terminals of the paraventricular nucleus of thalamus (PVT) NT neurons in the BLA enhanced reward learning and impaired punishment learning, while disruption of the PVT-BLA NTergic signaling produced the opposite behavioral effects. During the K99 phase of the award, the applicant will receive training on using CRISPR-Cas9 mediated gene manipulation to interrogate NTergic contributions without affecting glutamate and applying machine learning-based computer vision tools and classification algorithms to extract distinct behavioral motifs and decode neural correlates. This training will allow the applicant to investigate the specific roles of NT in valence assignment and coding properties of BLA neurons during valence processing. To test this, the applicant will examine the impact of the CRISPR-mediated knockdown of the Nt gene in BLA-projecting PVT neurons on reward and punishment learning (Aim 1) and the effects of the knockdown on valence encoding properties of BLA-NAc and BLA-CeM neurons during a reward and punishment discrimination task (Aim 2). After securing an independent research position, the applicant will begin the R00 phase of the award by combining these approaches gleaned from the K99 phase with applicant’s expertise in studying cocaine addiction with self- administration and punished reward-seeking paradigms, to investigate how PVT NTergic inputs modulate the encoding of compulsive cocaine-seeking (Aim 3). Together, the training and scientific advances that will be achieved through the completion of this project will allow the applicant to secure a tenure track faculty position in a top research university and provide critical pilot data necessary to prepare the first R01 application to study more distributed neural pathways that govern motivated behavior and compulsive drug-seeking.