Sensory experiences cause long-term modifications of neuronal circuits by modulating activity-dependent transcription programs that are vital for regulation of long-term synaptic plasticity and memory. However, it has not been possible to precisely determine the interaction between neuronal activity patterns and transcription factor activity. Here we present a technique using two-photon fluorescence lifetime imaging (2pFLIM) with new FRET biosensors to chronically image in vivo signaling of CREB, an activity-dependent transcription factor important for synaptic plasticity, at single-cell resolution. Simultaneous imaging of the red-shifted CREB sensor and GCaMP permitted exploration of how experience shapes the interplay between CREB and neuronal activity in the neocortex of awake mice. Dark rearing increased the sensitivity of CREB activity to Ca2+ elevations and prolonged the duration of CREB activation to more than 24 h in the visual cortex. This technique will allow researchers to unravel the transcriptional dynamics underlying experience-dependent plasticity in the brain.

中文翻译：

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小鼠大脑中神经元和CREB活性之间的耦合的体内成像。

感觉体验通过调节依赖于活动的转录程序对神经元回路的长期修饰，这对调节长期突触可塑性和记忆至关重要。但是，不可能精确地确定神经元活动模式和转录因子活动之间的相互作用。在这里，我们提出了一种使用双光子荧光寿命成像（2pFLIM）和新型FRET生物传感器的技术，以单细胞分辨率对CREB的体内信号进行长期成像，CREB是一种重要的突触可塑性，其依赖于活动的转录因子。红移的CREB传感器和GCaMP的同时成像允许探索经验如何塑造清醒小鼠新皮层中CREB和神经元活动之间的相互作用。暗饲养增加了视觉皮层中CREB活性对Ca2 +升高的敏感性，并将CREB激活的持续时间延长到24小时以上。这项技术将使研究人员能够揭示依赖于经验的大脑可塑性的转录动力学。