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

项目摘要 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相互连接，以 才能使大脑功能正常。过去的研究已经调查了由以下因素形成的连接率 体外培养神经元，这些细胞在自闭症小鼠模型(MeCP2 KO)中都显示出异常 小鼠)和精神分裂症患者的人类诱导多能干细胞(HiPSC)来源的神经元 (Brennand等人，2011年)。但简单地将皮质神经元分成兴奋性和抑制性两组 几乎没有触及皮质细胞类型的多样性的表面，已知的特异性之间的联系 异常或正常大脑中的特定细胞类型。在项目5中，我们建议开发并实现 新的工具将允许分析在HiPSC培养中形成的连接的细胞类型特异性- 衍生的神经元。这些检测是基于使用糖蛋白缺失的狂犬病病毒进行跨突触 连接神经元的标记和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年；Wall等人， 2010年)

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)