New Molecular Tools to Characterize Cortical Circuit Function in Non-Murine Mamma

表征非鼠妈妈皮层回路功能的新分子工具

基本信息

批准号：
8489440
负责人：
DAVID FITZPATRICK
金额：
$ 28.5万
依托单位：
MAX PLANCK FLORIDA CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8489440
关键词：
3&apos Untranslated Regions Address Adenoviruses Animal Mammary Glands Antibodies Brain Codon Nucleotides Development Diagnosis Disease Effectiveness Ferrets Foundations Genes Genetic Recombination Goals Health Image Imagery Immunohistochemistry Kinetics Knowledge Label Life Light Mammals Mediating Mental disorders Microscopy Molecular Monitor Mus Neurons Photons Population Primates Proteins Reporter Serotyping Specificity Structure Technology Testing Time Transgenes Viral Viral Vector Virus Visual Cortex cell type design expression vector helper-dependent adenoviral vector in vivo mouse development nervous system disorder neural circuit new technology novel strategies optogenetics promoter public health relevance recombinant viral vector recombinase research study response selective expression success tool transgene expression vector

项目摘要

DESCRIPTION (provided by applicant): New technologies that have made it possible to visualize specific classes of neurons, and to monitor and manipulate their activity in vivo, are transforming our understanding of neural circuit function and development. While the range of tools for molecular circuit visualization and manipulation continues to grow at an explosive rate in the mouse, the development of comparable tools for the study of circuits in non-murine mammals lags far behind. The inability to employ state of the art molecular tools for cell-type specific visualization and manipulation of neural circuits severely limits progress in understanding important aspects of cortical organization that are not well represented in the mouse brain. The goal of this application is to begin to bridge this technology gap by developing a recombinant viral vector for use in non-murine mammals that will permit in vivo expression of molecular reporters and effectors using cell-type specific promoters. As proof of principle, we will generate and characterize a helper-dependent adenovirus (HdAd) vector that will allow the selective expression of genetically encoded fluorescent proteins and light-sensitive channels in GABAergic neurons of the ferret visual cortex. The availability of a viral construct that yields expression selectively in GABAergic neurons of non-murine mammals will make it possible to address a host of questions that are critical for understanding cortical function in health and disease. Moreover, success in generating this construct will open the door to a battery of cell-type specific molecular tools that will be of tremendous value for the study of cortical and subcortical structures in a wide range of mammalian species including primates.

描述（由申请人提供）：新技术使特定类别的神经元可视化以及监测和操纵它们在体内的活动成为可能，正在改变我们对神经回路功能和发育的理解。虽然用于小鼠分子电路可视化和操作的工具范围继续以爆炸性速度增长，但用于研究非鼠类哺乳动物电路的类似工具的开发却远远落后。无法采用最先进的分子工具进行细胞类型特异性可视化和神经回路操作，严重限制了理解小鼠大脑中未得到很好体现的皮质组织重要方面的进展。该应用的目标是通过开发用于非鼠类哺乳动物的重组病毒载体来开始弥合这一技术差距，该载体将允许使用细胞类型特异性启动子在体内表达分子报告基因和效应子。作为原理证明，我们将生成并表征一种辅助依赖性腺病毒 (HdAd) 载体，该载体将允许在雪貂视觉皮层的 GABA 能神经元中选择性表达基因编码的荧光蛋白和光敏通道。在非鼠类哺乳动物的 GABA 能神经元中选择性表达的病毒构建体的出现，将有可能解决许多对于理解健康和疾病中的皮质功能至关重要的问题。此外，成功生成这种结构将为一系列细胞类型特异性分子工具打开大门，这些工具对于研究包括灵长类动物在内的多种哺乳动物的皮质和皮质下结构具有巨大价值。