Robust trans-synaptic labeling technologies for cell type-specific quantitation of synaptic connectivity

强大的跨突触标记技术，用于突触连接的细胞类型特异性定量

• 批准号: 9313719
• 负责人: EDWARD M CALLAWAY
• 金额: $ 32.88万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9313719
• 关键词: 16p11.2; 17p11.2; 7q11.23; Astrocytes; Autistic Disorder; Autopsy; Biological Assay; Brain; Cell Line; Cellular Assay; Cerebral cortex; Cholecystokinin; Collaborations; FMR1; Genotype; Glutamates; Glycoproteins; Haplotypes; Helper Viruses; Human; In Vitro; Infection; Interneurons; Knockout Mice; Label; Methods; Methyl-CpG-Binding Protein 2; Modeling; Mus; Neurons; Neurotransmitters; Pathway Analysis; Patients; Pattern; Point Mutation; Rabies; Rabies virus; Reagent; Reporter; Schizophrenia; Serotyping; Specificity; Subfamily lentivirinae; Surface; Synapses; System; TSC2 gene; Technology; Testing; Tissues; Viral Vector; Wild Type Mouse; autism spectrum disorder; base; biosignature; cell type; cellular transduction; excitatory neuron; human subject; in vivo; induced pluripotent stem cell; inhibitory neuron; interest; mouse model; neural circuit; new technology; novel; pre-clinical; tool

Project Summary Neural circuits are composed of networks of specific cell types that are interconnected in precise patterns to give rise to normal brain function. Past studies have investigated the rates of connections that are formed by cultured neurons in vitro and these have revealed abnormalities both in a mouse model of autism (MeCP2 KO mice) and for human induced pluripotent stem cell (hiPSC)-derived neurons from schizophrenic patients (Brennand et al., 2011). But simple separation of cortical neurons into just excitatory versus inhibitory groups barely scratches the surface of the diversity of cortical cell types, the known specificity of connections between specific cell types in the abnormal or normal brain. In Project 5 we propose to develop and then implement novel tools that will allow assays of the cell-type specificity of connections that are formed in cultures of hiPSC- derived neurons. These assays are based on the use of glycoprotein deleted rabies viruses for trans-synaptic labeling of connected neurons, and conditional expression of EnvA/TVA targeting systems for directing the initial infection of rabies virus to specific cell types (Wickersham et al., 2007b; Marshel et al., 2010; Wall et al., 2010)

项目概要 神经回路由特定细胞类型的网络组成，这些网络以精确的模式互连 产生正常的大脑功能。过去的研究调查了由以下因素形成的连接率 体外培养的神经元，这些结果揭示了自闭症小鼠模型（MeCP2 KO 小鼠）和来自精神分裂症患者的人类诱导多能干细胞（hiPSC）衍生的神经元 （布伦南德等人，2011）。但将皮质神经元简单地分为兴奋组和抑制组 仅仅触及皮层细胞类型多样性的表面，以及已知的皮层细胞类型之间连接的特异性 异常或正常大脑中的特定细胞类型。在项目5中，我们建议开发并实施 新工具将允许分析 hiPSC 培养物中形成的连接的细胞类型特异性 派生神经元。这些测定基于使用糖蛋白缺失的狂犬病病毒进行跨突触 连接神经元的标记，以及用于指导 EnvA/TVA 靶向系统的条件表达 狂犬病病毒对特定细胞类型的初始感染（Wickersham 等人，2007b；Marshel 等人，2010；Wall 等人， 2010）