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マウス体性感覚野を用いた皮質カラム構造の発達メカニズムの解明

利用小鼠体感皮层阐明皮质柱结构的发育机制

基本信息

批准号：
13J05789
负责人：
羅ブンジュウ
金额：
$ 1.41万
依托单位：
The Graduate University for Advanced Studies
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2013
资助国家：
日本
起止时间：
2013-04-01 至 2015-03-31
项目状态：
已结题

项目摘要

“Barrels” in layer 4 (L4) of mouse somatosensory cortex is a model system for the examination of cortical column development and function. They are established during early postnatal stage in an activity-dependent manner. To understand how neural activity modulates dendritic refinement and cell patterning of barrel cells, we developed vector systems termed "Supernova", which enables high Intensity single-cell labeling and labeled cell-specific gene-manipulation. Briefly, this system synergistically use the tTA/TRE expression system and site-directed recombination system (e.g. Cre/loxP) to enhance the expression of genes of interest only in a sparse population of cells. We demonstrated that multiple genes can be co-expressed in individual cells. And we showed that other types of site-specific recombination systems (e.g. flpe/FRT, Dre/Rox) can also be used. Moreover, we showed that Supernova systems can be used for RNA interference. Thus, the Supernova systems provide powerful tools in various aspects of neuroscience, for elucidation of the neural properties and gene functions in single cell levels.Using Supernova system, we performed morphological analysis of barrel cells’ dendrites during development, which may provide valuable information for elucidating the mechanisms of development of cortical neural circuit underlying pattern of columnar architecture.

小鼠体感皮层第4层（L4）中的“桶”是用于检查皮层柱发育和功能的模型系统。它们在出生后早期以活动依赖的方式建立。为了理解神经活动如何调节树突细化和桶细胞的细胞模式化，我们开发了称为“超新星”的载体系统，其能够实现高强度单细胞标记和标记的细胞特异性基因操作。简而言之，该系统协同使用tTA/TRE表达系统和定点重组系统（例如Cre/loXP）来增强仅在稀疏细胞群中感兴趣基因的表达。我们证明了多个基因可以在单个细胞中共表达。并且我们表明，也可以使用其他类型的位点特异性重组系统（例如flpe/FRT、Dre/Rox）。此外，我们表明超新星系统可用于RNA干扰。因此，Supernova系统为神经科学的各个方面提供了强有力的工具，可以在单细胞水平上阐明神经特性和基因功能。我们利用Supernova系统对发育过程中的桶形细胞树突进行了形态学分析，这可能为阐明以柱状结构模式为基础的皮层神经回路的发育机制提供有价值的信息。