Circuit-Specific Interrogation of the Primate Claustrum

灵长类动物闭状体的电路特异性询问

• 批准号: 10708856
• 负责人: Hala Galal El-Nahal
• 金额: $ 4.02万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2025-09-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10708856
• 关键词: Address; Animal Model; Animals; Appearance; Area; Attention; Back; Behavior; Behavioral; Brain; Brain Diseases; Cells; Cerebral cortex; Claustral structure; Cognition; Disease; Esthesia; Esthetics; Exhibits; Eye Movements; Functional Magnetic Resonance Imaging; Genes; Genetic Techniques; Geometry; Goals; Health; Histology; Human; Injections; Jaw; Label; Location; Macaca; Maps; Mediating; Methods; Monkeys; Motor; Movement; Neurons; Operative Surgical Procedures; Opsin; Optics; Outcome; Pain; Pathologic; Performance; Physiological; Prefrontal Cortex; Primates; Proteins; Psychophysics; Publishing; Rattus; Recombinant adeno-associated virus (rAAV); Research Design; Rodent; Role; Saccades; Sensorimotor functions; Shapes; Signal Transduction; Sleep; Specificity; Stimulus; Structure; System; Techniques; Technology; Testing; Thinness; Training; Translating; Viral; Viral Vector; Virus; Vision; Visual; Work; anatomical tracing; career; clinically relevant; cognitive function; decrease resilience; distraction; experimental study; extracellular; fluorophore; frontal eye fields; genetic approach; genetic technology; gray matter; nervous system disorder; neural; neurophysiology; neuropsychiatric disorder; nonhuman primate; novel strategies; optogenetics; visual cognition; visual motor

PROJECT SUMMARY The claustrum is a narrow, subcortical sheet of gray matter with interconnections across the cerebral cortex. Its irregular shape and deep location make it difficult to study with classical neurophysiological techniques. Recent genetic technologies such as optogenetics provide new hope for understanding the claustrum. They have advanced its study in rodents, implicating the claustrum in an array of sensorimotor and cognitive functions. Circuit-specific studies are needed to resolve how these functions map to the claustrum’s widespread connections. Such studies would be especially valuable in macaque monkeys, the animal model most homologous to humans. My overall goal is to translate viral technologies to monkeys and use them to study the role of a specific claustrum-prefrontal cortical circuit in visuo-saccadic behavior. During systematic testing of viral vectors in macaques, I identified a promising candidate for delivering opsin genes to the claustrum: the retrograde virus rAAV2-retro. When injected into the frontal eye field (FEF), a prefrontal cortical area involved in vision, movement, and cognition, rAAV2-retro constructs yielded strong labeling of FEF-projecting claustrum neurons. This finding provides a long-sought, unique opportunity to study claustrum neurons with circuit-level specificity in the primate brain. I propose to validate viral techniques in the claustro-FEF circuit and use them to test my overall hypothesis that the claustrum mediates competitive selection of the most behaviorally relevant stimulus via suppression of all other stimuli. To test my hypothesis, I have three integrated aims. In Aim 1, I will conduct anatomical tracing studies to elucidate the motif of connectivity between the claustrum and FEF. In Aim 2, I will use rAAV2-retro to express an inhibitory opsin in FEF-projecting claustrum neurons. Then I will use phototagging to identify those neurons and extracellularly record from them to characterize the signals sent from the claustrum to the FEF during a battery of saccade tasks. In Aim 3, I will use the same inhibitory opsins to selectively silence FEF-projecting claustrum neurons, allowing me to determine whether they are required for competitive selection of saccade targets. The claustrum is implicated in a wide range of neurological diseases and neuropsychiatric disorders. Thus, the results of the proposed work will help reveal how pathological changes to neural activity in the claustrum may contribute to brain disorders. In addition to its clinical relevance, the proposed work will significantly expand my doctoral training to include neural recording and optogenetic methods in non-human primates.

项目总结 屏状核是一层狭窄的皮质下灰质，跨大脑皮层相互连接。它的 不规则的形状和深部的位置使经典的神经生理学技术难以进行研究。近期 光遗传学等基因技术为理解屏蔽门提供了新的希望。他们有 在啮齿动物上推进了它的研究，表明屏蔽体与一系列感觉运动和认知功能有关。 需要特定电路的研究来解决这些功能如何映射到屏蔽门广泛的 关系。这类研究在猕猴身上尤其有价值，猕猴是最具动物模型的动物。 与人类同源。我的总体目标是将病毒技术转化为猴子，并用它们来研究 特定屏蔽区-前额叶皮质环路在视觉扫视行为中的作用。在对病毒进行系统测试期间 在猕猴体内，我发现了一种有希望将视蛋白基因转移到屏蔽层的候选基因： 逆行病毒rAAV2-retro。当注射到额叶眼场(FEF)时，前额叶皮质区域参与 视觉、运动和认知，rAAV2-retro结构产生了FEF投射屏蔽谱的强烈标记 神经元。这一发现为在电路水平上研究屏蔽神经元提供了一个长期寻求的独特机会 灵长类动物大脑中的专一性。我建议在CLaustro-FEF电路中验证病毒技术，并使用它们来 测试我的总体假设，即屏蔽门调节行为上最相关的竞争选择 通过抑制所有其他刺激来刺激。为了验证我的假设，我有三个综合目标。在《目标1》中，我将 进行解剖学追踪研究，以阐明屏骨和前额叶之间的连接主题。在AIM 2，我将使用rAAV2-retro在FEF投射的屏状核神经元中表达抑制性视蛋白。那我就会用 光标记识别这些神经元，并从细胞外记录它们，以表征从 在一系列扫视任务中向FEF发射的频谱。在目标3中，我将使用相同的抑制性视蛋白 选择性地使投射FEF的屏蔽神经元沉默，使我能够确定它们是否需要 对扫视目标的竞争性选择。屏蔽症与多种神经系统疾病有关。 和神经精神障碍。因此，拟议工作的结果将有助于揭示病理变化 屏蔽区的神经活动可能会导致大脑紊乱。除了它的临床意义外， 拟议的工作将大大扩展我的博士培训，将神经记录和光遗传学方法包括在内 在非人类灵长类动物中。