Determining how neurotensin mediates valence processing and compulsive cocaine seeking

项目摘要/摘要 物质使用障碍的一个特点是尽管有严重的不良后果，但仍继续使用药物。而当 对啮齿动物的研究表明，利用正价和负价处理都可以影响 这种强迫性的奖赏行为模式与惩罚相冲突，其背后的机制 调控价格过程和指导最终行为选择的机制尚不清楚。研究表明， 结果表明，杏仁基底外侧核(BLA)优先投射至伏核(BLA-NAC)。 编码正价，包括蔗糖和可卡因预测线索和驱动接近行为，而 杏仁中央核(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神经元可以编码价处理 在强迫性寻求可卡因期间，并在正在进行的行为中集体调解决策。这 拟议的项目旨在确定指导BLA-NAC之间价态分配的基本神经底物 和BLA-CEM神经元(K99时相)，并检查是否相同的机制参与了编码 强迫性寻求可卡因(R00阶段)。初步数据显示，神经降压素(NT)是一种13种氨基酸 多肽在调节BLA中的价态过程中起着关键作用。具体地说，激活 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 N能信号的中断产生相反的行为 效果。在奖励的K99阶段，申请者将接受使用CRISPR-Cas9中介的培训 在不影响谷氨酸和应用机的情况下询问神经功能的基因操作 基于学习的计算机视觉工具和分类算法，用于提取不同的行为主题和 破译神经相关基因。此培训将允许申请者调查NT在价态中的具体角色 价加工过程中BLA神经元的分配和编码特性。为了测试这一点，申请者将 检测CRISPR介导的BLA投射的PVT神经元NT基因敲除对大鼠脑脊液中神经营养因子的影响 奖惩学习(目标1)和击倒对价编码特性的影响 奖惩辨别任务中的BLA-NAC和BLA-CEM神经元(目标2)。确保安全后 作为一个独立的研究职位，申请人将通过结合以下几个方面开始奖项的R00阶段 从K99阶段收集的方法与申请者在研究可卡因成瘾方面的专业知识和自我 管理和惩罚性奖赏寻求范式，以调查PVT神经输入如何调节 强迫性寻求可卡因的编码(目标3)。总之，培训和科学进步将是 通过完成这一项目，申请人将获得终身教职。 在一流研究型大学，并提供必要的关键试点数据，以准备第一个R01应用程序进行研究 支配动机行为和强迫性药物寻求的神经通路更加分散。

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.