Abstract
Optogenetic methods have emerged as powerful tools for dissecting neural circuit connectivity, function and dysfunction. We used a bacterial artificial chromosome (BAC) transgenic strategy to express the H134R variant of channelrhodopsin-2, ChR2(H134R), under the control of cell type–specific promoter elements. We performed an extensive functional characterization of the newly established VGAT-ChR2(H134R)-EYFP, ChAT-ChR2(H134R)-EYFP, Tph2-ChR2(H134R)-EYFP and Pvalb(H134R)-ChR2-EYFP BAC transgenic mouse lines and demonstrate the utility of these lines for precisely controlling action-potential firing of GABAergic, cholinergic, serotonergic and parvalbumin-expressing neuron subsets using blue light. This resource of cell type–specific ChR2(H134R) mouse lines will facilitate the precise mapping of neuronal connectivity and the dissection of the neural basis of behavior.
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Acknowledgements
We thank P. Miao, K. Harley, L. Strickland and J. Chemla for technical assistance with mouse husbandry and genotyping, Q. Liu and members of the NeuroTransgenic lab at Duke University for pronuclear injections of BAC DNA and other members of the Feng laboratory for their support, C. Keller-McGandy for help with histology in the Graybiel lab, and J. Ren and other members of the Luo lab for providing electrophysiology expertise and input. This work was supported by an American Recovery and Reinvestment Act grant from the US National Institute of Mental Health (RC1-MH088434) to G.F., a National Alliance for Research on Schizophrenia and Depression: The Brain and Behavior Research Foundation Young Investigator award and US National Institutes of Health Ruth L. Kirschstein National Research Service award (F32MH084460) to J.T.T. and a National Institute of Mental Health grant to A.M.G. (R01 MH060379).
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G.F., K.D. and G.J.A. initiated the project. K.D. provided ChR2(H134R) DNA constructs. S.Z., L.Q. and B.G. generated the ChR2 BAC transgenic founder lines. S.Z. and L.Q. screened the founder lines. S.Z. performed all confocal imaging experiments. J.T.T. performed electrophysiological recordings, and analyzed and interpreted acute-brain-slice experiments for all mouse lines. J.T. performed electrophysiological recordings, and M.L. and J.T. analyzed and interpreted acute brain slice experiments on ChAT-ChR2(H134R)-EYFP line 6 and VGAT-ChR2(H134R)-EYFP line 8 mice. H.E.A. performed in vivo electrophysiology, and H.E.A. and A.M.G. analyzed and interpreted in vivo electrophysiology data on ChAT-ChR2(H134R)-EYFP line 6 mice. J.T.T. and G.F. wrote the manuscript.
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Zhao, S., Ting, J., Atallah, H. et al. Cell type–specific channelrhodopsin-2 transgenic mice for optogenetic dissection of neural circuitry function. Nat Methods 8, 745–752 (2011). https://doi.org/10.1038/nmeth.1668
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DOI: https://doi.org/10.1038/nmeth.1668
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