Department of Physiology, UT Southwestern Medical Center, Dallas, TX, USA.
Department of Neuroscience, Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA.
Nat Neurosci. 2023 Jul;26(7):1295-1307. doi: 10.1038/s41593-023-01357-w. Epub 2023 Jun 12.
Neural activity is modulated over different timescales encompassing subseconds to hours, reflecting changes in external environment, internal state and behavior. Using Drosophila as a model, we developed a rapid and bidirectional reporter that provides a cellular readout of recent neural activity. This reporter uses nuclear versus cytoplasmic distribution of CREB-regulated transcriptional co-activator (CRTC). Subcellular distribution of GFP-tagged CRTC (CRTC::GFP) bidirectionally changes on the order of minutes and reflects both increases and decreases in neural activity. We established an automated machine-learning-based routine for efficient quantification of reporter signal. Using this reporter, we demonstrate mating-evoked activation and inactivation of modulatory neurons. We further investigated the functional role of the master courtship regulator gene fruitless (fru) and show that fru is necessary to ensure activation of male arousal neurons by female cues. Together, our results establish CRTC::GFP as a bidirectional reporter of recent neural activity suitable for examining neural correlates in behavioral contexts.
神经活动在不同的时间尺度上被调节,包括亚秒到小时,反映了外部环境、内部状态和行为的变化。我们以果蝇为模型,开发了一种快速且双向的报告基因,为最近的神经活动提供了细胞水平的读出。该报告基因使用 CREB 调节的转录共激活因子 (CRTC) 的核质分布。GFP 标记的 CRTC(CRTC::GFP)的亚细胞分布在数分钟内双向变化,反映了神经活动的增加和减少。我们建立了一种基于自动化机器学习的常规方法,用于有效地量化报告基因信号。使用该报告基因,我们证明了交配诱发的调节神经元的激活和失活。我们进一步研究了主要求偶调节基因 fruitless (fru) 的功能作用,并表明 fru 对于确保雄性唤醒神经元被雌性线索激活是必要的。总之,我们的结果确立了 CRTC::GFP 作为最近神经活动的双向报告基因,适用于在行为背景下研究神经相关性。