A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing

Allen Institute for Brain Science, Seattle, Washington, USA.
Nature Neuroscience (Impact Factor: 16.1). 03/2012; 15(5):793-802. DOI: 10.1038/nn.3078
Source: PubMed


Cell type-specific expression of optogenetic molecules allows temporally precise manipulation of targeted neuronal activity. Here we present a toolbox of four knock-in mouse lines engineered for strong, Cre-dependent expression of channelrhodopsins ChR2-tdTomato and ChR2-EYFP, halorhodopsin eNpHR3.0 and archaerhodopsin Arch-ER2. All four transgenes mediated Cre-dependent, robust activation or silencing of cortical pyramidal neurons in vitro and in vivo upon light stimulation, with ChR2-EYFP and Arch-ER2 demonstrating light sensitivity approaching that of in utero or virally transduced neurons. We further show specific photoactivation of parvalbumin-positive interneurons in behaving ChR2-EYFP reporter mice. The robust, consistent and inducible nature of our ChR2 mice represents a significant advance over previous lines, and the Arch-ER2 and eNpHR3.0 mice are to our knowledge the first demonstration of successful conditional transgenic optogenetic silencing. When combined with the hundreds of available Cre driver lines, this optimized toolbox of reporter mice will enable widespread investigations of neural circuit function with unprecedented reliability and accuracy.

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    • "In both the cortex and hippocampus we found Na v b1- containing AIS that were likely to belong to inhibitory neurons based on their differential orientation relative to the majority of AIS observed. To confirm this observation and to detail our investigation of Na v b1 expression in inhibitory neurons, we used three mouse strains engineered to express tdTomato in parvalbumin (PV)-, somatostatin (SST)-, and vasoactive intestinal polypeptide (VIP)-positive inhibitory neuron subclasses (Fig. 7; Hippenmeyer et al., 2005; Madisen et al., 2012; Taniguchi et al., 2011). Na v b1 staining was seen in essentially all PV and SST inhibitory neurons but in only a fraction of VIP-positive interneurons. "
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    • "All experiments were conducted in accordance with the National Institutes of Health guidelines regarding the care and use of animals and were approved by the Duke University Institutional Animal Care and Use Committee (Protocol Number: A027- 14-02). For behavioral testing, male Ai32 mice expressing a floxed STOP cassette upstream of the ChR2(H134R)-EYFP gene (Madisen et al., 2012) were bred with dopamine D 1 receptor Cre (D1-Cre) mice to yield D1-ChR2 mice that selectively expressed the light-gated cation channel, ChR2, in D1-expressing neurons (n = 3; aged 4–7 months). Controls were D1-Cre mice "
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