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.

项目概要 屏状核是一片狭窄的皮层下灰质，与大脑皮层相互连接。 不规则的形状和深部位置使得用经典的神经生理学技术进行研究变得困难。 光遗传学等遗传技术为理解屏状体提供了新的希望。 推进了对啮齿类动物的研究，将屏状体与一系列感觉运动和认知功能联系起来。 需要针对特定​​电路的研究来解决这些功能如何映射到屏状核的广泛分布 这种联系对于猕猴（最重要的动物模型）尤其有价值。 我的总体目标是将病毒技术转化为猴子并用它们来研究 在病毒的系统测试过程中，特定的屏状核-前额皮质回路在视觉扫视行为中的作用。 在猕猴体内的载体中，我发现了一种将视蛋白基因传递到屏状体的有希望的候选者： 逆行病毒 rAAV2-retro 当注射到额叶眼区 (FEF) 时，涉及前额皮质区域。 视觉、运动和认知，rAAV2-retro 构建体产生了 FEF 投射屏状体的强标记 这一发现为研究屏状神经元的电路级提供了一个长期寻求的独特机会。 我建议验证幽闭-FEF 回路中的病毒技术，并将其用于 检验我的总体假设，即屏状核介导对行为最相关的竞争性选择 为了检验我的假设，我在目标 1 中设定了三个综合目标。 进行解剖追踪研究以阐明屏状体和 FEF 之间的连接基序 In Aim。 2，我将使用 rAAV2-retro 在 FEF 投影屏状神经元中表达抑制性视蛋白，然后我将使用。 光标记来识别这些神经元并在细胞外记录它们以表征从神经元发送的信号 在一系列眼跳任务中，我将使用相同的抑制性视蛋白来识别屏状核到 FEF。 选择性地沉默 FEF 投射的屏状神经元，使我能够确定它们是否需要 眼跳目标的竞争性选择与多种神经系统疾病有关。 因此，拟议工作的结果将有助于揭示病理变化。 除了其临床相关性之外，屏状核中的神经活动可能会导致大脑疾病。 拟议的工作将显着扩展我的博士培训，包括神经记录和光遗传学方法 在非人类灵长类动物中。