Resource | Published:

Cell type–specific channelrhodopsin-2 transgenic mice for optogenetic dissection of neural circuitry function

Nature Methods volume 8, pages 745752 (2011) | Download Citation

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Targeting and readout strategies for fast optical neural control in vitro and in vivo. J. Neurosci. 27, 14231–14238 (2007).

  2. 2.

    et al. Sparse optical microstimulation in barrel cortex drives learned behaviour in freely moving mice. Nature 451, 61–64 (2008).

  3. 3.

    et al. Optical activation of lateral amygdala pyramidal cells instructs associative fear learning. Proc. Natl. Acad. Sci. USA 107, 12692–12697 (2010).

  4. 4.

    , , , & Neural substrates of awakening probed with optogenetic control of hypocretin neurons. Nature 450, 420–424 (2007).

  5. 5.

    et al. Ectopic expression of a microbial-type rhodopsin restores visual responses in mice with photoreceptor degeneration. Neuron 50, 23–33 (2006).

  6. 6.

    et al. Light-induced rescue of breathing after spinal cord injury. J. Neurosci. 28, 11862–11870 (2008).

  7. 7.

    , & AGRP neurons are sufficient to orchestrate feeding behavior rapidly and without training. Nat. Neurosci. 14, 351–355 (2011).

  8. 8.

    , , & FLEX switch targets channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping. J. Neurosci. 28, 7025–7030 (2008).

  9. 9.

    et al. Driving fast-spiking cells induces gamma rhythm and controls sensory responses. Nature 459, 663–667 (2009).

  10. 10.

    , , , & Optical deconstruction of parkinsonian neural circuitry. Science 324, 354–359 (2009).

  11. 11.

    et al. Genetic dissection of an amygdala microcircuit that gates conditioned fear. Nature 468, 270–276 (2010).

  12. 12.

    et al. Regulation of parkinsonian motor behaviours by optogenetic control of basal ganglia circuitry. Nature 466, 622–626 (2010).

  13. 13.

    , , & Parvalbumin neurons and gamma rhythms enhance cortical circuit performance. Nature 459, 698–702 (2009).

  14. 14.

    , , , & Dopaminergic terminals in the nucleus accumbens but not the dorsal striatum corelease glutamate. J. Neurosci. 30, 8229–8233 (2010).

  15. 15.

    et al. Phasic firing in dopaminergic neurons is sufficient for behavioral conditioning. Science 324, 1080–1084 (2009).

  16. 16.

    et al. Cholinergic interneurons control local circuit activity and cocaine conditioning. Science 330, 1677–1681 (2010).

  17. 17.

    , , & Pathway-specific feedforward circuits between thalamus and neocortex revealed by selective optical stimulation of axons. Neuron 65, 230–245 (2010).

  18. 18.

    et al. Fast synaptic subcortical control of hippocampal circuits. Science 326, 449–453 (2009).

  19. 19.

    et al. In vivo light-induced activation of neural circuitry in transgenic mice expressing channelrhodopsin-2. Neuron 54, 205–218 (2007).

  20. 20.

    et al. High-speed mapping of synaptic connectivity using photostimulation in Channelrhodopsin-2 transgenic mice. Proc. Natl. Acad. Sci. USA 104, 8143–8148 (2007).

  21. 21.

    et al. Visual properties of transgenic rats harboring the channelrhodopsin-2 gene regulated by the thy-1.2 promoter. PLoS ONE 4, e7679 (2009).

  22. 22.

    , , & Activation of groups of excitatory neurons in the mammalian spinal cord or hindbrain evokes locomotion. Nat. Neurosci. 13, 246–252 (2010).

  23. 23.

    , , , & Non-redundant odor coding by sister mitral cells revealed by light addressable glomeruli in the mouse. Nat. Neurosci. 13, 1404–1412 (2010).

  24. 24.

    , , & Functional connectome of the striatal medium spiny neuron. J. Neurosci. 31, 1183–1192 (2011).

  25. 25.

    et al. Cloning of a functional vesicular GABA and glycine transporter by screening of genome databases. FEBS Lett. 417, 177–183 (1997).

  26. 26.

    The loading of neurotransmitters into synaptic vesicles. Biochimie 82, 327–337 (2000).

  27. 27.

    et al. Ultrafast optogenetic control. Nat. Neurosci. 13, 387–392 (2010).

  28. 28.

    & Neurochemical phenotypes of the afferent and efferent projections of the mouse medial habenula. Neuroscience 161, 827–837 (2009).

  29. 29.

    , & Intrinsic membrane properties underlying spontaneous tonic firing in neostriatal cholinergic interneurons. J. Neurosci. 20, 8493–8503 (2000).

  30. 30.

    & Electrophysiological and pharmacological characterization of serotonergic dorsal raphe neurons recorded extracellularly and intracellularly in rat brain slices. Brain Res. 289, 109–119 (1983).

  31. 31.

    , & Somatodendritic autoreceptor regulation of serotonergic neurons: dependence on L-tryptophan and tryptophan hydroxylase-activating kinases. Eur. J. Neurosci. 21, 945–958 (2005).

  32. 32.

    , & Heterogeneity of firing properties among rat thalamic reticular nucleus neurons. J. Physiol. (Lond.) 582, 195–208 (2007).

  33. 33.

    & A novel T-type current underlies prolonged Ca(2+)-dependent burst firing in GABAergic neurons of rat thalamic reticular nucleus. J. Neurosci. 12, 3804–3817 (1992).

  34. 34.

    & Electrophysiological properties of in vitro Purkinje cell dendrites in mammalian cerebellar slices. J. Physiol. (Lond.) 305, 197–213 (1980).

  35. 35.

    , , , & The columnar and laminar organization of inhibitory connections to neocortical excitatory cells. Nat. Neurosci. 14, 100–107 (2011).

  36. 36.

    et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).

  37. 37.

    et al. Fluorescent labeling of newborn dentate granule cells in GAD67-GFP transgenic mice: a genetic tool for the study of adult neurogenesis. PLoS ONE 5, e12506 (2010).

  38. 38.

    et al. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28, 41–51 (2000).

  39. 39.

    et al. Shank3 mutant mice display autistic-like behaviours and striatal dysfunction. Nature 472, 437–442 (2011).

  40. 40.

    , , , & Ascorbate inhibits edema in brain slices. J. Neurochem. 74, 1263–1270 (2000).

  41. 41.

    , & HEPES prevents edema in rat brain slices. Neurosci. Lett. 303, 141–144 (2001).

  42. 42.

    et al. Habenula “cholinergic” neurons corelease glutamate and acetylcholine and activate postsynaptic neurons via distinct transmission modes. Neuron 69, 445–452 (2011).

  43. 43.

    et al. Tetrode technology: advances in implantable hardware, neuroimaging, and data analysis techniques. J. Neurosci. Methods 117, 141–152 (2002).

  44. 44.

    et al. Stable encoding of task structure coexists with flexible coding of task events in sensorimotor striatum. J. Neurophysiol. 102, 2142–2160 (2009).

  45. 45.

    , , , & Activity of striatal neurons reflects dynamic encoding and recoding of procedural memories. Nature 437, 1158–1161 (2005).

Download references

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).

Author information

Author notes

    • Shengli Zhao
    •  & Jonathan T Ting

    These authors contributed equally to this work.

Affiliations

  1. Department of Neurobiology, Duke University Medical Center, Durham, North Carolina, USA.

    • Shengli Zhao
    • , Jonathan T Ting
    • , Li Qiu
    • , Bernd Gloss
    •  & Guoping Feng
  2. McGovern Institute for Brain Research and Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Jonathan T Ting
    • , Hisham E Atallah
    • , Ann M Graybiel
    •  & Guoping Feng
  3. National Institute of Biological Sciences, Beijing, China.

    • Jie Tan
    •  & Minmin Luo
  4. Duke NeuroTransgenic Laboratory, Duke University Medical Center, Durham, North Carolina, USA.

    • Bernd Gloss
  5. Center for Functional Connectomics, Korea Institute of Science and Technology, Seoul, Republic of Korea.

    • George J Augustine
  6. Program in Neuroscience and Behavioral Disorders, Duke–National University of Singapore Graduate Medical School, Singapore.

    • George J Augustine
  7. Agency for Science, Technology and Research, Duke–National University of Singapore Neuroscience Research Partnership, Singapore.

    • George J Augustine
  8. Department of Bioengineering, Stanford University, California, USA.

    • Karl Deisseroth

Authors

  1. Search for Shengli Zhao in:

  2. Search for Jonathan T Ting in:

  3. Search for Hisham E Atallah in:

  4. Search for Li Qiu in:

  5. Search for Jie Tan in:

  6. Search for Bernd Gloss in:

  7. Search for George J Augustine in:

  8. Search for Karl Deisseroth in:

  9. Search for Minmin Luo in:

  10. Search for Ann M Graybiel in:

  11. Search for Guoping Feng in:

Contributions

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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Guoping Feng.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–15, Supplementary Tables 1–3

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nmeth.1668

Further reading