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

小鼠躯体感觉皮层第四层(L4)的“桶”是检查皮质柱发育和功能的模型系统。它们是在出生后早期以依赖活动的方式建立的。为了了解神经活动如何调节桶状细胞的树突状细化和细胞构型，我们开发了名为“超新星”的载体系统，它能够实现高强度的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的目的基因的表达。我们证明了多个基因可以在单个细胞中共表达。我们还证明了其他类型的位点特异性重组系统(如Flpe/FRT，DRE/Rox)也可以使用。此外，我们还证明了超新星系统可以用于RNA干扰。因此，超新星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。利用超新星系统，我们对发育过程中桶状细胞的树突进行了形态分析，这可能为阐明柱状结构模式下皮层神经回路的发育机制提供有价值的信息。

“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.