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: 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
• 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' 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

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，并在体内监测和操纵它们的活动成为可能，正在改变我们对神经回路功能和发育的理解。虽然用于分子电路可视化和操作的工具范围在小鼠中继续以爆炸式的速度增长，但用于非小鼠哺乳动物电路研究的可比工具的发展远远落后。无法使用最先进的分子工具来进行细胞类型特异性的可视化和神经回路的操作，严重限制了在理解皮层组织的重要方面方面的进展，这些方面在小鼠大脑中没有得到很好的体现。本应用程序的目标是通过开发用于非小鼠哺乳动物的重组病毒载体，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能神经元中选择性地表达遗传编码的荧光蛋白和光敏感通道。在非小鼠哺乳动物的gaba能神经元中选择性表达的病毒构建体的可用性，将使解决对理解健康和疾病中的皮质功能至关重要的一系列问题成为可能。此外，这种结构的成功生成将为一系列细胞类型特异性分子工具打开大门，这些工具将对包括灵长类动物在内的广泛哺乳动物物种的皮层和皮层下结构的研究具有巨大价值。