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

• 批准号: 8616378
• 负责人: DAVID FITZPATRICK
• 金额: $ 23.28万
• 依托单位: MAX PLANCK FLORIDA CORPORATION
• 依托单位国家: 美国
• 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/8616378
• 关键词: 3' 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; 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; two-photon; vector

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）：已有可能可视化特定类别神经元并监测和操纵其在体内活动的新技术正在改变我们对神经电路功能和发育的理解。尽管用于小鼠的分子电路可视化和操纵工具的范围继续以爆炸性的速度增长，但开发了可比的工具，用于研究非乳腺哺乳动物中的电路落后。无法使用艺术分子工具来进行细胞类型的特定可视化和对神经回路的操纵，严重限制了在小鼠大脑中无法很好地表示皮质组织的重要方面的进展。该应用的目的是通过开发重组病毒载体来开始弥合该技术差距，以用于非乳腺哺乳动物中，该哺乳动物将允许使用细胞类型的特定启动子在体内表达分子记者和效应子。作为原理的证明，我们将生成和表征辅助腺病毒（HDAD）载体，该腺病毒（HDAD）载体将允许在雪貂视觉皮层的GABA能神经元中选择性地表达遗传编码的荧光蛋白和光敏通道。病毒构建体的可用性在非乳腺哺乳动物的GABA能神经元中有选择地表达，这将使解决一系列对于理解健康和疾病中皮质功能至关重要的问题。此外，生成这种构建体的成功将为一系列细胞类型的特定分子工具打开大门，这对于在包括灵长类动物在内的广泛哺乳动物物种中研究皮质和皮层结构具有巨大价值。