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

描述（由申请人提供）：使用转基因工具选择性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性，这在神经可塑性研究中具有重大影响。近年来，已经开发了许多分子工具来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在不同神经元细胞类型和回路中的应用受到限制，特别是因为开发每个单独的转基因动物所需的时间和精力。这个问题已经通过投入大量精力开发细胞类型特异性Cre-driver线而得到部分解决。然而，在创建必要数量的Cre响应性转基因系中仍然存在需要克服的障碍，所述Cre响应性转基因系能够以足够强的水平表达分子工具，用于成像和以细胞类型特异性的亚细胞靶向方式进行功能操作，以供学术研究人员使用。为了尝试解决这个问题，我们提出系统地靶向一组基因组位点，这些基因组位点将分别在抑制性和兴奋性神经元中传达基因的强普遍表达或选择性突触前和突触后特异性靶向，以在这些基因组位点中开发Cre应答器平台，任何现有或新开发的分子工具都可以容易地插入其中。我们致力于利用该计划为学术界提供多样化的转基因小鼠，这些小鼠将利用许多新的工具和探针来研究神经网络，特别是研究神经元可塑性，因为它与行为和生理学有关。艾伦研究所拥有高通量ISH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待开发的转基因品系相结合，我们将能够增加转基因小鼠作为研究神经可塑性的方法的实用性，使这些实验动物免费提供给神经科学界。公共卫生相关性：神经可塑性是大脑发育和大脑获得知识和技能的能力的基础过程，也是许多大脑疾病的组成部分。遗传工具已广泛应用于塑性研究的各个领域。我们拟议的研究将为神经科学界提供一个转基因平台，将最先进的探针和工具用于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理解可塑性在大脑功能和疾病中的作用产生重大影响，从而找到治疗疾病和改善公共健康的方法。

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.