Huang Z Josh, Yu Wenjiang, Lovett Chanel, Tonegawa Susumu
Howard Hughes Medical Institute, RIKEN-MIT Neuroscience Research Center, Departments of Biology, and Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Genesis. 2002 Mar;32(3):209-17. doi: 10.1002/gene.10054.
A new strategy for visualizing neuronal cell morphology of mouse brain based on Cre/loxP recombination-activated gene expression is described. A "reporter" transgenic line was generated which expressed a fusion gene encoding a dendrite-targeted green fluorescent protein (MAP2-GFP) upon deletion of a transcription/translation STOP (transcription and translation stop signal) cassette. Cre transgenic "deleter" lines were established that activated reporter gene expression at various frequencies in pyramidal neurons in the forebrain. A deleter line was identified which activated a MAP2-GFP reporter gene at very low frequency (less than 0.1% of pyramidal neurons) and allowed the visualization of dendritic structures of individual neocortical and hippocampal pyramidal neurons. In addition, vertical "columns" of pyramidal neurons in the neocortex were labeled in these mice. In a second deleter line, a MAP2-GFP reporter gene was selectively activated in pyramidal neurons of the CA-1 subregion of the hippocampus in young mice. With its combinatorial property, this binary recombination-activated neuronal marker system should facilitate the study of detailed morphology, connectivity, and plasticity of defined classes of live neurons in vitro and in vivo.
描述了一种基于Cre/loxP重组激活基因表达来可视化小鼠脑神经元细胞形态的新策略。构建了一个“报告基因”转基因系,其在转录/翻译终止(转录和翻译终止信号)盒缺失时表达编码树突靶向绿色荧光蛋白(MAP2-GFP)的融合基因。建立了Cre转基因“删除者”系,其在前脑锥体神经元中以不同频率激活报告基因表达。鉴定出一个删除者系,其以非常低的频率(少于0.1%的锥体神经元)激活MAP2-GFP报告基因,并能可视化单个新皮质和海马锥体神经元的树突结构。此外,这些小鼠的新皮质中锥体神经元的垂直“柱”被标记。在第二个删除者系中,年轻小鼠海马CA-1亚区的锥体神经元中选择性激活了MAP2-GFP报告基因。凭借其组合特性,这种二元重组激活的神经元标记系统应有助于在体外和体内研究特定类型活神经元的详细形态、连接性和可塑性。