A transgenic platform to produce genetic tools for monitoring & manipulating plas

描述(申请人提供)：使用转基因工具选择性地标记和检测单个神经元群体，并调节这些细胞中的基因表达以操纵活动，可以在神经可塑性研究中产生重大影响。近年来，许多分子工具被开发出来，用于研究神经回路和可塑性的精细细节和控制。然而，这些工具在细胞和亚细胞水平上的不同神经细胞类型和电路中的应用一直受到限制，特别是因为开发每个单独的转基因动物需要时间和精力。这一问题已经通过投入大量精力开发特定单元类型的CRE驱动器线而得到部分解决。然而，在建立必要数量的Cre响应型转基因株系以足够强的水平表达分子工具以进行成像和以细胞类型特异的亚细胞靶向方式进行功能操纵以供学术研究人员使用方面，仍有障碍需要克服。为了解决这个问题，我们建议系统地定位于一组基因组座位，这些座位将分别在抑制性和兴奋性神经元中表达强烈的普遍表达或选择性的突触前和突触后特异性基因，以在这些基因组座位上开发一个Cre-Responder平台，任何现有的或新开发的分子工具都可以很容易地插入到这些基因组座位上。我们致力于利用这一计划为学术界提供多样化的转基因小鼠，以利用许多新的工具和探测器来研究神经网络，特别是研究与行为和生理相关的神经元可塑性。艾伦研究所拥有一个高吞吐量的平台和高分辨率图像采集和数据库的能力。通过将这些创新技术与将要开发的转基因品系相结合，我们将能够通过向神经科学界免费提供这样的实验动物来增加转基因小鼠作为研究神经可塑性的方法的实用性。公共卫生相关性：神经可塑性是大脑发育和大脑获取知识和技能能力的基本过程，也是许多大脑疾病的组成部分。基因工具已广泛应用于塑性研究的各个领域。我们提议的研究将为神经科学界提供一个转基因平台，以结合最先进的探针和工具，在细胞和亚细胞水平上监测和操纵可塑性。神经科学界使用这些遗传工具将对进一步了解可塑性在大脑功能和疾病中所起的作用产生重大影响，从而找到治疗疾病和改善公众健康的方法。

DESCRIPTION (provided by applicant): The use of transgenic tools to selectively label and detect individual neuronal populations and modulate gene expression in those cells to manipulate activity can have significant impact in neuroplasticity research. In recent years a number of molecular tools have been developed to study neural circuitry and plasticity at exquisite detail and control. However, application of these tools in different neuronal cell types and circuits at cellular and subcellular level has been limited, in particular because of the time and effort needed to develop each individual transgenic animal. This issue has been partially addressed by putting in great effort into the development of cell type specific Cre-driver lines. However, there have still been hurdles to overcome in creating the necessary numbers of Cre responsive transgenic lines that can express the molecular tools at sufficiently strong levels for imaging and for functional manipulation in a cell type specific, subcellularly targeted manner for use by academic researchers. To attempt to address this issue, we propose to systematically target a set of genomic loci that will convey strong ubiquitous expression or selective pre and post-synaptic specific targeting of genes in inhibitory and excitatory neurons respectively, to develop a Cre-responder platform in these genomic loci to which any existing or newly developed molecular tools can be easily inserted. We are committed to use this program to provide diverse transgenic mice to the academic community that will exploit many of the new tools and probes to study neural networks and in particular study neuronal plasticity as it relates to behavior and physiology. The Allen Institute has a high throughput ISH platform and capability of high resolution image acquisition and data-basing. By combining these innovative technologies with the transgenic lines to be developed, we will be able to increase the utility of the transgenic mice as an approach to study neuroplasticity by making such experimental animals freely available for the neuroscience community. PUBLIC HEALTH RELEVANCE: Neuroplasticity is a fundamental process underlying brain development and brain's ability to acquire knowledge and skills, as well as a component of many brain disorders. Genetic tools have been widely used in every area of plasticity research. Our proposed research will provide to the neuroscience community a transgenic platform to incorporate state-of-the-art probes and tools for monitoring and manipulating plasticity at cellular and subcellular levels. Use of these genetic tools by the neuroscience community will have major impact on the further understanding of the roles of plasticity plays in brain function and diseases, leading to ways in treating diseases and improving public health.