Investigating the role of Tsc1 in neocortical circuit assembly

研究 Tsc1 在新皮质电路组装中的作用

• 批准号: 8717098
• 负责人: Laura Anne DeNardo
• 金额: $ 4.71万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8717098
• 关键词: Affect; Autistic Disorder; Behavior; Behavior Control; Behavioral; Behavioral Symptoms; Benign; Brain; Brain region; Cells; Comb animal structure; Complex; Decision Making; Development; Disease; Epilepsy; Equilibrium; Exhibits; Foundations; Future; Gene Mutation; Genes; Genetic; Hippocampus (Brain); Intervention; Investigation; Knock-out; Lead; Link; Maps; Medial; Molecular; Mono-S; Motor output; Mus; Mutation; Neocortex; Neurodevelopmental Disorder; Neurons; Pathway interactions; Patients; Pattern; Phenotype; Prefrontal Cortex; Rabies; Rabies virus; Regulation; Research; Role; Sensory; Short-Term Memory; Signal Pathway; Signal Transduction; Social Interaction; Structure; Synapses; System; Techniques; Testing; Tuberous sclerosis protein complex; Tumor Suppressor Genes; Work; base; cell type; in vivo; information processing; knockout gene; mTOR protein; multisensory; neocortical; neural circuit; public health relevance; sensorimotor system; tumor

DESCRIPTION (provided by applicant): The mammalian neocortex is a six-layered structure that creates a representation of the world by integrating sensory information and controls behavior by generating the appropriate motor output. Each layer processes information differently, so understanding how layer-specific neuronal networks are organized will provide a foundation for understanding how different layers function within circuits. The medial prefrontal cortex (mPFC) integrates information from many brain regions and is involved in complex behaviors including social interaction and decision-making. Importantly, mPFC circuits are often disrupted in neurodevelopmental disorders such as autism, but little is known about the fine scale organization of mPFC or how mPFC layers integrate different kinds of information. The Luo lab recently developed a modified rabies-based mono-trans- synaptic tracing technique that allows for detailed analysis of local circuitry in addition to whole brain long- distance mapping. This new rabies technique can be combined with layer-specific Cre driver mouse lines to generate detailed maps of layer-specific circuits in medial prefrontal cortex (mPFC). Cre-dependent rabies tracing will also be combined with cell-type specific gene knockout, so these maps will provide a basis for studying genetic regulation of neocortical connectivity in development and disease with unprecedented precision and scope. To begin to dissect the molecular pathways involved in establishing specific mPFC connectivity, this project will focus on the role of the autism-related gene Tsc1. Recent studies showed that Tsc1 deletion increases excitatory synaptic connectivity and alters the balance of excitation and inhibition, causing hyperexcitability in hippocampal neurons. This phenotype may be related to the role of Tsc1 in Tuberous Sclerosis Complex, a disease in which patients suffer from benign tumors, epilepsy, and autism. Performing Cre-dependent rabies tracing in conditional Tsc1fl/fl mice crossed with layer-specific Cre-drivers will facilitate investigation of the cell-autonomous role o Tsc1 in the development of mPFC connectivity and layer- specific organization. As Tsc1 is a negative regulator of the mammalian target of rapamycin complex, mTORC1, the molecular mechanisms underlying the in vivo function of Tsc1 will also be investigated to elucidate how these signaling pathways regulate brain development. This work will provide a deeper understanding of the molecular and the circuit-level mechanisms that give rise to autism and epilepsy in patients with Tsc1 mutations.

描述（由申请人提供）：哺乳动物新皮层是一个六层结构，通过整合感觉信息来创建对世界的表征，并通过产生适当的运动输出来控制行为。每一层都以不同的方式处理信息，因此了解特定于层的神经元网络是如何组织的，将为理解不同层在电路中的功能提供基础。内侧前额叶皮层（mPFC）整合来自许多大脑区域的信息，并参与包括社交和决策在内的复杂行为。重要的是，mPFC电路在神经发育障碍（如自闭症）中经常被破坏，但人们对mPFC的精细尺度组织或mPFC层如何整合不同类型的信息知之甚少。Luo实验室最近开发了一种改良的基于狂犬病的单跨突触追踪技术，该技术除了全脑长距离映射外，还可以对局部电路进行详细分析。这种新的狂犬病技术可以与特定于层的Cre驱动小鼠线相结合，以生成内侧前额叶皮层（mPFC）中特定于层的电路的详细地图。依赖Cre的狂犬病追踪也将与细胞类型特异性基因敲除相结合，因此这些图谱将以前所未有的精度和范围为研究发育和疾病中新皮层连接的遗传调控提供基础。为了开始剖析参与建立特定mPFC连接的分子通路，本项目将重点关注自闭症相关基因Tsc 1的作用。最近的研究表明，Tsc 1缺失增加了兴奋性突触连接，改变了兴奋和抑制的平衡，导致海马神经元过度兴奋。这种表型可能与Tsc 1在恶性硬化综合征中的作用有关，该疾病是一种患者患有良性肿瘤、癫痫和自闭症的疾病。在与层特异性Cre驱动因子杂交的条件性Tsc 1fl/fl小鼠中进行Cre依赖性狂犬病追踪将有助于研究Tsc 1在mPFC连接性和层特异性组织发育中的细胞自主作用。由于Tsc 1是雷帕霉素复合物mTORC 1的哺乳动物靶点的负调节剂，因此还将研究Tsc 1体内功能的分子机制，以阐明这些信号通路如何调节大脑发育。这项工作将提供一个更深入的了解分子和电路水平的机制，引起自闭症和癫痫患者的Tsc 1突变。