Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Nat Methods. 2013 Sep;10(9):889-95. doi: 10.1038/nmeth.2559. Epub 2013 Jul 14.
Identifying the neuronal ensembles that respond to specific stimuli and mapping their projection patterns in living animals are fundamental challenges in neuroscience. To this end, we engineered a synthetic promoter, the enhanced synaptic activity-responsive element (E-SARE), that drives neuronal activity-dependent gene expression more potently than other existing immediate-early gene promoters. Expression of a drug-inducible Cre recombinase downstream of E-SARE enabled imaging of neuronal populations that respond to monocular visual stimulation and tracking of their long-distance thalamocortical projections in living mice. Targeted cell-attached recordings and calcium imaging of neurons in sensory cortices revealed that E-SARE reporter expression correlates with sensory-evoked neuronal activity at the single-cell level and is highly specific to the type of stimuli presented to the animals. This activity-dependent promoter can expand the repertoire of genetic approaches for high-resolution anatomical and functional analysis of neural circuits.
确定对特定刺激作出反应的神经元集合并在活体动物中绘制它们的投射模式是神经科学的基本挑战。为此，我们设计了一种合成启动子，即增强的突触活动反应元件（E-SARE），它比其他现有的即刻早期基因启动子更有效地驱动神经元活动依赖性基因表达。E-SARE 下游的药物诱导型 Cre 重组酶的表达使我们能够对单眼视觉刺激作出反应的神经元群体进行成像，并在活体小鼠中追踪它们的长距离丘脑皮质投射。对感觉皮层中神经元的靶向细胞贴附记录和钙成像显示，E-SARE 报告基因的表达与单细胞水平的感觉诱发神经元活动相关，并且高度特异性地针对动物所接受的刺激类型。这种活动依赖性启动子可以扩展遗传方法的范围，用于对神经回路进行高分辨率的解剖学和功能分析。
