Light-induced silencing of neural activity in Rosa26 knock-in and BAC transgenic mice conditionally expressing the microbial halorhodopsin eNpHR3.

Itaru Imayoshi, Sawako Tabuchi, Mami Matsumoto, Satsuki Kitano, Hitoshi Miyachi, Ryoichiro Kageyama, Akihiro Yamanaka

Scientific reports 2020 Feb 21

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An engineered light-inducible chloride pump, Natronomonas pharaonis halorhodopsin 3 (eNpHR3) enables temporally and spatially precise inhibition of genetically defined cell populations in the intact nervous tissues. In this report, we show the generation of new mouse strains that express eNpHR3-EYFP fusion proteins after Cre- and/or Flp-mediated recombination to optically inhibit neuronal activity. In these mouse strains, Cre/Flp recombination induced high levels of opsin expression. We confirmed their light-induced activities by brain slice whole-cell patch clamp experiments. eNpHR3-expressing neurons were optically hyperpolarized and silenced from firing action potentials. In prolonged silencing of action potentials, eNpHR3 was superior to eNpHR2, a former version of the engineered pump. Thus, these eNpHR3 mouse strains offer reliable genetic tools for light-induced inhibiting of neuronal activity in defined sets of neurons.

Citation

Itaru Imayoshi, Sawako Tabuchi, Mami Matsumoto, Satsuki Kitano, Hitoshi Miyachi, Ryoichiro Kageyama, Akihiro Yamanaka. Light-induced silencing of neural activity in Rosa26 knock-in and BAC transgenic mice conditionally expressing the microbial halorhodopsin eNpHR3. Scientific reports. 2020 Feb 21;10(1):3191