Imaging neuromodulation in the brain

大脑神经调节成像

• 批准号: 10543730
• 负责人: David J Anderson
• 金额: $ 39.17万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10543730
• 关键词: Address; Adult; Arousal; Behavior; Biogenic Amines; Biological Assay; Brain; Calcium; Cells; Courtship; Data; Decision Making; Dopamine; Drosophila genus; Emotions; Enzymes; FOS gene; Fluorescent in Situ Hybridization; Genetic; Genetic Models; Goals; Grant; Homologous Gene; Image; Immediate-Early Genes; In Situ Hybridization; Insecta; Interneurons; Label; Learning; Link; Mediating; Morphology; Motivation; Mushroom Bodies; Nature; Negative Valence; Neuromodulator; Neurons; Neuropeptides; Neurotransmitters; Norepinephrine; Octopamine; Odors; Phylogeny; Play; Population; Positive Valence; Publications; Rewards; Rodent; Role; Self Administration; Social Behavior; Social Controls; Stimulus; System; Techniques; Testing; Time; addiction; base; candidate identification; conditioning; experimental study; male; model organism; neural circuit; neurobiological mechanism; neuropeptide F; neuroregulation; optogenetics; preference; psychostimulant; response; social

Addiction involves brain systems mediating internal states of motivation, arousal and reward, as well as emotions. Such internal states influence goal-directed behaviors and decision-making. A common feature of such internal states is their valence and their persistence: they can have a positive or negative valence, and can outlast their triggering stimulus for many minutes. However the neurobiological mechanisms that underlie the persistence of internal states, and their relationship to the encoding of valence, are poorly understood. Drosophila provides a tractable genetic model organism for studying how neuromodulators act on neural circuits to control persistent internal states that govern goal-directed behavior and decision-making. We have discovered that P1 interneurons, which control male courtship behavior, can when activated promote a persistent internal state of social arousal or motivation, which can last for minutes. In a publication supported by the base grant, we have obtained evidence of a link between P1 interneurons and neurons that respond to octopamine (OA), an insect homolog of norepinephrine (NE), which is known to facilitate psychostimulant self- administration in rodents. We have also identified a downstream target of P1 neurons, called pCd cells, which appear to play a key requisite role in determining the persistence of an internal state of social arousal. During the extension period, we will continue our studies of how P1 neurons promote a rewarding internal state, and the relationship of these mechanisms to the positive valence, or rewarding nature, of P1 stimulation. In the first 2 years, we will focus on pursuing Aims 3 and 4 of the base grant. These aims were: Aim 3) to test the hypothesis that P1 neuron activation is positively valenced and rewarding; Aim 4) to investigate neuromodulatory mechanisms involved in P1 reward learning. In unpublished experiments, we have discovered that activation of P1 neurons can produce a real-time place preference (RTPP), and that it can also serve as an unconditional stimulus (US) for conditioned olfactory preference (COP). Both of these findings indicate that P1 activation is positively valenced, and that it can be rewarding. We plan to investigate whether plasticity during COP occurs at or downstream of P1 neurons, and whether P1 neurons are necessary for expression of the COP (Aim 3). Preliminary experiments suggest that dopamine (DA) may play a role in modulating the effects effects of P1 stimulation. We will confirm and extend these findings, and also investigate the role(s) of other neuromodulators including biogenic amines such as octopamine (OA), which we have shown to modulate the effect of P1 stimulation to activate aSP2 neurons that control social behavior9. Furthermore, we will investigate whether mushroom body (MB) neurons involved in reward learning are also involved in P1-mediated odor conditioning (Aim 4). Given previous data, we expect to find a role for the MB, but precisely which subset of MB neurons are involved is not clear. In Merit Extension Aim 5, we will investigate the role of other neuromodulators in P1-induced persistent social arousal and reward learning. Candidate neuromodulatory targets of P1 neurons include serotonergic (Trh+) neurons, a subset of which is activated in response to P1 stimulation (preliminary results), and neuropeptide F (NPF), which has been implicated in reward in other contexts. We will approach this problem using functional connectomics, in which optogenetic activation of P1 neurons is combined with calcium imaging in populations containing putative neuromodulatory targets of these cells. Target neurons can be “filled” using photo-activatable GFP (PA-GFP), and their morphology used as a “search image” to identify specific genetic drivers that label that subset of cells. Using these drivers, activation and silencing of these neurons can be performed in the context of both P1-mediated reward (RTPP and COP assays), and persistent social arousal. We have successfully established this approach and used it to identify pCd neurons, which are persistently activated by P1 neurons and required for persistent social behaviors triggered by P1 activation. As a complementary approach, we will take advantage of recent advances that we have made in techniques for whole-mount fluorescent in situ hybridization (FISH) in the adult brain, which allow identification of candidate follower cells activated by optogenetic stimulation of P1 neurons using FISH probes for hr38, an immediate early gene (analogous to c-fos) in Drosophila. Double-label FISH can be performed using hr38 and probes for neurotransmitter biosynthetic enzymes or neuropeptides, to identify neuromodulators expressed in P1 targets. A fundamental question is whether the mechanism mediating P1-induced persistent activity is also involved in reward. To address this question, in Merit Extension Aim 6 we will investigate the role of pCd neurons in P1- mediated reward learning, using functional manipulations of these cells. Preliminary data suggest that persistent activation of pCd neurons by P1 cells is modulated by DA, and we will investigate how this modulatory influence is exerted. We anticipate that these experiments will yield general principles of how persistent internal reward states are encoded by brains, with potentially broad relevance across phylogeny.

成瘾涉及介导内部动机，唤醒和奖励的大脑系统，以及 情绪。这种内部状态会影响目标指导的行为和决策。一个共同特征的 这种内部状态是它们的价和持久性：它们可以具有正价或负面价，并且 可以超过其触发刺激数分钟。但是，基于的神经生物学机制 内部状态的持久性及其与价的编码的关系知之甚少。 果蝇提供了一种可探讨的遗传模型生物，用于研究神经调节剂如何作用于神经元素 控制持续的内部状态的电路，这些状态管理目标指导的行为和决策。我们有 发现控制男性求爱行为的P1中间神经元可以在激活时促进 持续的社会唤醒或动机的内部状态可以持续几分钟。在支持的出版物中 通过基础赠款，我们获得了P1中间神经元与神经元之间有联系的证据 章鱼胺（OA），一种去甲肾上腺素（NE）的绝缘同源物，已知可以促进精神刺激性自我 啮齿动物的管理。我们还确定了P1神经元的下游靶标，称为PCD细胞，该靶点 似乎在确定社会唤醒的内部状态的持久性方面起着关键的必要作用。 在扩展期间，我们将继续研究P1神经元如何促进有意义的内部 状态以及这些机制与P1刺激的正价或奖励性质的关系。 在最初的两年中，我们将专注于基本赠款的目标3和4。这些目的是：目标3）测试 P1神经元激活的假设是积极的价值和奖励。目标4）调查 P1奖励学习涉及的神经调节机制。在未发表的实验中，我们有 发现P1神经元的激活可以产生实时位置偏好（RTPP），并且也可以 作为条件嗅觉偏好（COP）的无条件刺激（US）。这两个发现 表明P1激活具有积极的价值，并且可能是有益的。我们计划调查是否 COP期间的可塑性发生在P1神经元的或下游，以及P1神经元是否需要 COP的表达（AIM 3）。初步实验表明多巴胺（DA）可能在 调节P1刺激的影响。我们将确认并扩展这些发现，并调查 其他神经调节剂的作用，包括生物胺，例如章鱼胺（OA），我们已经拥有 显示以调节P1刺激的效果激活控制社会行为的ASP2神经元。 此外，我们将调查参与奖励学习的肌肉室（MB）神经元是否也是 参与P1介导的气味调节（AIM 4）。鉴于以前的数据，我们希望找到MB的作用 但是，尚不清楚涉及哪些MB神经元子集。 在功绩扩展目标5中，我们将研究其他神经调节剂在P1引起的持续性中的作用 社会唤醒和奖励学习。 P1神经元的候选神经调节靶标包括血清素能 （TRH+）神经元，其中一个子集是根据P1模拟（初步结果）而激活的，并且 神经肽F（NPF），在其他情况下已暗示。我们将解决这个问题 使用功能连接组，其中P1神经元的光遗传激活与钙成像结合 在包含这些细胞推定的神经调节靶标的种群中。靶神经元可以使用 照片活跃的GFP（PA-GFP）及其形态用作“搜索图像”来识别特定的通用 标记该细胞子集的驱动器。使用这些驱动程序，这些神经元的激活和沉默可以是 在P1介导的奖励（RTPP和COP分析）和持续的社会唤醒的背景下进行。 我们已经成功建立了这种方法，并用它来识别PCD神经元，这是持续的 由P1神经元激活，并需要由P1激活触发的持续社会行为。作为 完成的方法，我们将利用我们在技术方面取得的最新进步 成人大脑中的全部荧光原位杂交（FISH），允许识别候选者 随着P1神经元的光遗传学刺激使用鱼类问题，HR38激活的追随者细胞，即 果蝇的早期基因（类似于c-fos）。可以使用HR38进行双标签鱼和 神经递质生物合成酶或神经肽，以鉴定在P1靶标中表达的神经调节剂。 一个基本问题是介导P1引起的持续活动的机制是否也参与 报酬。为了解决这个问题，在绩效延伸目标6中，我们将研究PCD神经元在P1-的作用 使用这些细胞的功能操纵，介导的奖励学习。初步数据表明 P1细胞对PCD神经元的持续激活由DA调节，我们将研究如何 调节作用被施加。我们预计这些实验将产生有关如何 持续的内部奖励状态由大脑编码，在系统发育中可能具有广泛的相关性。