Egr1-EGFP transgenic mouse allows in vivo recording of Egr1 expression and neural activity
Egr1-EGFP transgenic mouse allows in vivo recording of Egr1 expression and neural activity
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DOI：
10.1016/j.jneumeth.2021.109350
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发表时间：
2021-09
影响因子：
3
通讯作者：
Guangyu Wang;Hong Xie;Yi-ling Hu;Qi-Nan Chen;Chenhui Liu;Kaiyuan Liu;Yuze Yan;J. Guan
中科院分区：
文献类型：
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作者：
Guangyu Wang;Hong Xie;Yi-ling Hu;Qi-Nan Chen;Chenhui Liu;Kaiyuan Liu;Yuze Yan;J. Guan
即刻早期基因(IEGs)因其对刺激的快速反应而被认为是活跃神经元的标志物。随着GENSAT项目对IEG-EGFP记者的开发，IEG的应用得到了极大的扩展。新方法以Egr1-EGFP转基因小鼠为例，在对其荧光信号进行详细验证的基础上，对Egr1-EGFP报告系统的使用提出了改进。Egr1-EGFP信号在刺激前后3小时的变化与刺激诱导的神经元活动呈正相关。有趣的是，刺激后阶段的持续神经元活动模式也与刺激诱导的Egr1-EGFP信号变化相关。此外，丰富的环境参与了戏剧性的神经元激活，允许详细描述EGF1-EGFP的表达动态。与现有的方法(S)相比，人们习惯于根据IEG-EGFP报告系统的原始荧光信号来推断神经元的活动，这一方法受到EGFP和IEG之间不同的蛋白质调控或动力学特性的强烈阻碍。结论综上所述，本工作证明了EGR1-EGFP信号与任务诱发的神经活动之间存在微弱但显著的相关性，并给出了信号动力学的详细描述。这不仅为理解IEG-EGFP荧光信号提供了依据，也为正确设计IEG-EGFP报告系统的实验设计提供了指导。
BackgroundImmediate-early genes (IEGs) have been serving as markers of active neurons for their rapid responses to stimulation. With the development of IEG-EGFP reporters by the GENSAT project, application of the IEGs have been greatly expanded. However, detailed validations for these systems are still lacking, causing trouble in the interpretation of the fluorescence signals.New methodIn this work, taken Egr1-EGFP transgenic mice as an example, we proposed an improvement for the usage of the Egr1-EGFP reporter system based on detailed validation of its fluorescence signals.ResultsFirstly, the exogenousEGFPmRNA levels were linearly correlated with the endogenousEgr1mRNA levels in neurons. Secondly, the 3-hr-changes of the Egr1-EGFP signals before and after the stimulus were positively correlated with the stimulus-induced neuronal activities. Interestingly, persistent neuronal activity patterns in the post-stimulus phase also showed correlation with the stimulus-induced Egr1-EGFP signal changes. Furthermore, enriched environments engaged dramatic neuronal activations, allowing detailed characterization of Egr1-EGFP expression dynamics.Comparison with existing method(s)People used to infer the neuronal activities based on the raw fluorescence signals of IEG-EGFP reporter system, which was strongly obstructed by distinct protein regulation or dynamic properties between the EGFP and the IEGs. We demonstrated a better way for data analysis and experimental design.ConclusionsTaken together, this work proves that Egr1-EGFP signal is weakly but significantly correlated to task-induced neural activity and gives detailed characterization of the signal dynamics. It not only provides basis for the understanding of the IEG-EGFP fluorescence signals but also offers instructions for proper experimental design with IEG-EGFP reporter systems.