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All-optical interrogation of neural circuits in behaving mice

View ORCID ProfileLloyd E. Russell, View ORCID ProfileHenry W. P. Dalgleish, View ORCID ProfileRebecca Nutbrown, View ORCID ProfileOliver M. Gauld, Dustin Herrmann, Mehmet Fişek, View ORCID ProfileAdam M. Packer, View ORCID ProfileMichael Häusser
doi: https://doi.org/10.1101/2021.06.29.450430
Lloyd E. Russell
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
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  • ORCID record for Lloyd E. Russell
Henry W. P. Dalgleish
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
2Sainsbury Wellcome Centre, University College London, London UK
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Rebecca Nutbrown
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
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Oliver M. Gauld
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
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Dustin Herrmann
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
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Mehmet Fişek
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
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Adam M. Packer
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
3Department of Physiology, Anatomy and Genetics, University of Oxford, UK
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  • For correspondence: adampacker@gmail.com m.hausser@ucl.ac.uk
Michael Häusser
1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK
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  • For correspondence: adampacker@gmail.com m.hausser@ucl.ac.uk
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Abstract

Recent advances combining two-photon calcium imaging and two-photon optogenetics with digital holography now allow us to read and write neural activity in vivo at cellular resolution with millisecond temporal precision. Such “all-optical” techniques enable experimenters to probe the impact of functionally defined neurons on neural circuit function and behavioural output with new levels of precision. This protocol describes the experimental strategy and workflow for successful completion of typical all-optical interrogation experiments in awake, behaving head-fixed mice. We describe modular procedures for the setup and calibration of an all-optical system, the preparation of an indicator and opsin-expressing and task-performing animal, the characterization of functional and photostimulation responses and the design and implementation of an all-optical experiment. We discuss optimizations for efficiently selecting and targeting neuronal ensembles for photostimulation sequences, as well as generating photostimulation response maps from the imaging data that can be used to examine the impact of photostimulation on the local circuit. We demonstrate the utility of this strategy using all-optical experiments in three different brain areas – barrel cortex, visual cortex and hippocampus – using different experimental setups. This approach can in principle be adapted to any brain area for all-optical interrogation experiments to probe functional connectivity in neural circuits and for investigating the relationship between neural circuit activity and behaviour.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/llerussell/Naparm

  • https://github.com/llerussell/STAMovieMaker

  • https://github.com/llerussell/SLMTransformMaker3D

  • https://github.com/llerussell/SLMPhaseMaskMaker3D

  • https://github.com/llerussell/Bruker_PrairieLink

  • https://github.com/llerussell/PyBehaviour

  • https://github.com/hwpdalgleish/TPBS

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 01, 2021.
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All-optical interrogation of neural circuits in behaving mice
Lloyd E. Russell, Henry W. P. Dalgleish, Rebecca Nutbrown, Oliver M. Gauld, Dustin Herrmann, Mehmet Fişek, Adam M. Packer, Michael Häusser
bioRxiv 2021.06.29.450430; doi: https://doi.org/10.1101/2021.06.29.450430
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All-optical interrogation of neural circuits in behaving mice
Lloyd E. Russell, Henry W. P. Dalgleish, Rebecca Nutbrown, Oliver M. Gauld, Dustin Herrmann, Mehmet Fişek, Adam M. Packer, Michael Häusser
bioRxiv 2021.06.29.450430; doi: https://doi.org/10.1101/2021.06.29.450430

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