A Flp-dependent G-CaMP9a transgenic mouse for neuronal imaging in vivo

用于体内神经元成像的 Flp 依赖性 G-CaMP9a 转基因小鼠

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作者:Masayuki Sakamoto, Masatoshi Inoue, Atsuya Takeuchi, Shigetaka Kobari, Tatsushi Yokoyama, Shin-Ichiro Horigane, Sayaka Takemoto-Kimura, Manabu Abe, Kenji Sakimura, Masanobu Kano, Kazuo Kitamura, Hajime Fujii, Haruhiko Bito

Abstract

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. In vivo 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 in vivo.

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