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

描述(申请人提供)：新技术使可视化特定类别的神经元成为可能，并在体内监测和操纵它们的活动，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纵的工具范围在小鼠中继续以爆炸性的速度增长，但用于研究非小鼠哺乳动物的电路的可比工具的开发远远落后。无法使用最先进的分子工具对神经回路进行细胞类型的特定可视化和操作，严重限制了在理解小鼠大脑中没有很好代表的皮质组织的重要方面方面的进展。这项应用的目标是通过开发一种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的特定启动子在体内表达分子报告器和效应器。作为原则的证明，我们将产生并鉴定一种辅助依赖型腺病毒(HdAd)载体，该载体将允许在雪貂视皮层的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一系列问题成为可能，这些问题对于理解健康和疾病中的皮质功能至关重要。此外，成功构建这种结构将为一系列细胞类型的特定分子工具打开大门，这些工具将对研究包括灵长类在内的多种哺乳动物的皮质和皮质下结构具有巨大价值。

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.