一种依赖 Flp 的 G-CaMP9a 转基因小鼠，用于神经元成像。
A Flp-dependent G-CaMP9a transgenic mouse for neuronal imaging .
机构信息
Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Hongo7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan.
Department of Optical Neural and Molecular Physiology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8507, Japan.
出版信息
Cell Rep Methods. 2022 Feb 14;2(2):100168. doi: 10.1016/j.crmeth.2022.100168. eCollection 2022 Feb 28.
Genetically encoded calcium indicators (GECIs) are widely used to measure calcium transients in neuronal somata and processes, and their use enables the determination of action potential temporal series in a large population of neurons. Here, we generate a transgenic mouse line expressing a highly sensitive green GECI, G-CaMP9a, in a Flp-dependent manner in excitatory and inhibitory neuronal subpopulations downstream of a strong CAG promoter. Combining this reporter mouse with viral or mouse genetic Flp delivery methods produces a robust and stable G-CaMP9a expression in defined neuronal populations without detectable detrimental effects. two-photon imaging reveals spontaneous and sensory-evoked calcium transients in excitatory and inhibitory ensembles with cellular resolution. Our results show that this reporter line allows long-term, cell-type-specific investigation of neuronal activity with enhanced resolution in defined populations and facilitates dissecting complex dynamics of neural networks
基因编码钙指示剂 (GECIs) 被广泛用于测量神经元胞体和突起中的钙瞬变，其使用能够确定大群体神经元中的动作电位时间序列。在这里，我们生成了一种转基因小鼠品系，该品系以 Flp 依赖性方式在兴奋性和抑制性神经元亚群中表达高度敏感的绿色 GECI，G-CaMP9a，该基因受强 CAG 启动子的调控。将这种报告小鼠与病毒或小鼠遗传 Flp 递送方法相结合，可在没有可检测到的有害影响的情况下，在特定神经元群体中产生强大且稳定的 G-CaMP9a 表达。双光子成像以细胞分辨率揭示兴奋性和抑制性集合中的自发和感觉诱发钙瞬变。我们的结果表明，该报告品系允许在定义的群体中以增强的分辨率进行长期的、细胞类型特异性的神经元活动研究，并有助于剖析神经网络的复杂动态。
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