Simultaneous two-photon optogenetics and imaging of cortical circuits in three dimensions

Abstract

The simultaneously imaging and manipulating of neural activity in three-dimensions could enable the functional dissection of neural circuits. Here we have combined two-photon optogenetics with simultaneous volumetric two-photon calcium imaging to manipulate neural activity in mouse neocortex in vivo in 3D, while maintaining cellular resolution. Using a hybrid holographic approach, we simultaneously photostimulate more than 80 neurons over 150 μm in depth in cortical layer 2/3 from mouse visual cortex. We validate the usefulness of the microscope by photoactivating in 3D selected groups of interneurons, suppressing the response of nearby pyramidal neurons to visual stimuli. Our all-optical method could be used as a general platform to read and write activity of neural circuits.

Article and author information

Author details

  1. Weijian Yang

    Department of Biological Sciences, Columbia University, New York, United States
    For correspondence
    wejyang@ucdavis.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0941-3496
  2. Luis Carrillo-Reid

    Department of Biological Sciences, Columbia University, New York, United States
    Competing interests
    No competing interests declared.
  3. Yuki Bando

    Department of Biological Sciences, Columbia University, New York, United States
    Competing interests
    No competing interests declared.
  4. Darcy S Peterka

    Department of Biological Sciences, Columbia University, New York, United States
    Competing interests
    Darcy S Peterka, is listed as an inventor of the following patent: Devices.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7351-5820
  5. Rafael Yuste

    Department of Biological Sciences, Columbia University, New York, United States
    For correspondence
    rmy5@columbia.edu
    Competing interests
    Rafael Yuste, is listed as an inventor of the following patent: Devices.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4206-497X

Funding

National Eye Institute (DP1EY024503)

  • Rafael Yuste

National Institute of Mental Health (R44MH109187)

  • Darcy S Peterka

Defense Advanced Research Projects Agency (N66001-15-C-4032)

  • Rafael Yuste

National Institute of Mental Health (R01MH100561)

  • Rafael Yuste

National Eye Institute (R21EY027592)

  • Darcy S Peterka

National Institute of Mental Health (R01MH101218)

  • Rafael Yuste

Defense Advanced Research Projects Agency (W91NF-14-1-0269)

  • Rafael Yuste

Army Research Laboratory (W911NF-12-1-0594)

  • Rafael Yuste

Army Research Office (W911NF-12-1-0594)

  • Rafael Yuste

Burroughs Wellcome Fund (1015761)

  • Weijian Yang

Uehara Memorial Foundation

  • Yuki Bando

National Eye Institute (R01EY011787)

  • Rafael Yuste

National Institute of Mental Health (R41MH100895)

  • Rafael Yuste

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Karel Svoboda, Janelia Research Campus, Howard Hughes Medical Institute, United States

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of Columbia University [protocol ID: AC-AAAM5100, AC-AAAM7951].

Version history

  1. Received: October 10, 2017
  2. Accepted: February 5, 2018
  3. Accepted Manuscript published: February 7, 2018 (version 1)
  4. Version of Record published: March 1, 2018 (version 2)

Copyright

© 2018, Yang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 19,583
    views
  • 2,585
    downloads
  • 168
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Weijian Yang
  2. Luis Carrillo-Reid
  3. Yuki Bando
  4. Darcy S Peterka
  5. Rafael Yuste
(2018)
Simultaneous two-photon optogenetics and imaging of cortical circuits in three dimensions
eLife 7:e32671.
https://doi.org/10.7554/eLife.32671

Share this article

https://doi.org/10.7554/eLife.32671

Further reading

    1. Neuroscience
    Nicola Masala, Manuel Mittag ... Tony Kelly
    Research Article

    Genetically encoded calcium indicators (GECIs) such as GCaMP are invaluable tools in neuroscience to monitor neuronal activity using optical imaging. The viral transduction of GECIs is commonly used to target expression to specific brain regions, can be conveniently used with any mouse strain of interest without the need for prior crossing with a GECI mouse line, and avoids potential hazards due to the chronic expression of GECIs during development. A key requirement for monitoring neuronal activity with an indicator is that the indicator itself minimally affects activity. Here, using common adeno-associated viral (AAV) transduction procedures, we describe spatially confined aberrant Ca2+ microwaves slowly travelling through the hippocampus following expression of GCaMP6, GCaMP7, or R-CaMP1.07 driven by the synapsin promoter with AAV-dependent gene transfer in a titre-dependent fashion. Ca2+ microwaves developed in hippocampal CA1 and CA3, but not dentate gyrus nor neocortex, were typically first observed at 4 wk after viral transduction, and persisted up to at least 8 wk. The phenomenon was robust and observed across laboratories with various experimenters and setups. Our results indicate that aberrant hippocampal Ca2+ microwaves depend on the promoter and viral titre of the GECI, density of expression, as well as the targeted brain region. We used an alternative viral transduction method of GCaMP which avoids this artefact. The results show that commonly used Ca2+-indicator AAV transduction procedures can produce artefactual Ca2+ responses. Our aim is to raise awareness in the field of these artefactual transduction-induced Ca2+ microwaves, and we provide a potential solution.

    1. Neuroscience
    John J Stout, Allison E George ... Amy L Griffin
    Research Article

    Functional interactions between the prefrontal cortex and hippocampus, as revealed by strong oscillatory synchronization in the theta (6–11 Hz) frequency range, correlate with memory-guided decision-making. However, the degree to which this form of long-range synchronization influences memory-guided choice remains unclear. We developed a brain-machine interface that initiated task trials based on the magnitude of prefrontal-hippocampal theta synchronization, then measured choice outcomes. Trials initiated based on strong prefrontal-hippocampal theta synchrony were more likely to be correct compared to control trials on both working memory-dependent and -independent tasks. Prefrontal-thalamic neural interactions increased with prefrontal-hippocampal synchrony and optogenetic activation of the ventral midline thalamus primarily entrained prefrontal theta rhythms, but dynamically modulated synchrony. Together, our results show that prefrontal-hippocampal theta synchronization leads to a higher probability of a correct choice and strengthens prefrontal-thalamic dialogue. Our findings reveal new insights into the neural circuit dynamics underlying memory-guided choices and highlight a promising technique to potentiate cognitive processes or behavior via brain-machine interfacing.