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: 10604913
• 负责人: Hala Galal El-Nahal
• 金额: $ 3.92万
• 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/10604913
• 关键词: Address; Animal Model; Animals; Appearance; Area; 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; Movement; Neurons; Operative Surgical Procedures; Opsin; Optics; Outcome; Pathologic; Performance; Physiological; Primates; Proteins; Psychophysics; Rattus; Research Design; Rodent; Role; Saccades; Sensorimotor functions; Shapes; Signal Transduction; Specificity; Stimulus; Structure; System; Techniques; Technology; Testing; Thinness; Training; Translating; Viral; Viral Vector; Virus; Vision; Visual; Work; anatomical tracing; base; career; clinically relevant; cognitive function; experimental study; extracellular; fluorophore; frontal eye fields; genetic approach; genetic technology; gray matter; nervous system disorder; neurophysiology; neuropsychiatric disorder; nonhuman primate; novel strategies; optogenetics; relating to nervous system; resilience; 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.

项目摘要 屏状核是一个狭窄的，皮层下的灰质层，与大脑皮层相互连接。其 不规则的形状和深的位置使得难以用经典的神经生理学技术进行研究。最近 光遗传学等遗传技术为了解屏状核提供了新的希望。他们有 在啮齿类动物中的研究取得了进展，表明屏状核与一系列感觉运动和认知功能有关。 需要对特定的回路进行研究，以解决这些功能如何映射到屏状核的广泛分布。 连接.这些研究在猕猴身上尤其有价值， 与人类同源。我的总体目标是将病毒技术转化到猴子身上，并用它们来研究 特定的闭孔-前额叶皮层回路在视觉扫视行为中的作用。在病毒的系统测试期间， 在猕猴中的载体，我确定了一个有希望的候选人提供视蛋白基因的屏状核： 逆行病毒rAAV 2-retro。当注射到额叶眼区（FEF）时，前额叶皮质区参与了 在视觉、运动和认知方面，rAAV 2-retro构建体产生了FEF投射屏状体的强标记 神经元这一发现提供了一个长期寻求的，独特的机会，研究屏状核神经元与电路水平 灵长类大脑的特异性我建议在幽闭FEF回路中验证病毒技术，并使用它们来 测试我的总体假设，即屏状体介导的竞争性选择最相关的行为 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，以阐明屏状体和FEF之间的连接基序。在Aim中 2、利用rAAV 2-retro在屏状核FEF投射神经元中表达抑制性视蛋白。那我就用 光标记来识别这些神经元，并从它们中进行细胞外记录，以表征从 在一系列扫视任务中屏状核到FEF。在目标3中，我将使用相同的抑制性视蛋白， 选择性地沉默FEF投射屏状核神经元，使我能够确定它们是否是 竞争性扫视目标选择。屏状核与多种神经系统疾病有关 和神经精神疾病。因此，拟议工作的结果将有助于揭示病理变化如何 屏状核的神经活动可能会导致大脑紊乱。除了临床意义外， 拟议的工作将大大扩展我的博士培训，包括神经记录和光遗传学方法 在非人类灵长类动物中。