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Integration of optogenetics with complementary methodologies in systems neuroscience

Key Points

  • Modern optogenetics enables temporally precise, acute or chronic, excitatory or inhibitory modulation of neuronal activity with cell type and anatomical specificity that can be tuned to timing and magnitude of naturally occurring patterns within the same experimental subject.

  • Diverse opsin variants exhibit unique spectral and kinetic features that are specifically suited for distinct experimental requirements.

  • Optogenetics can be used in combination with electrophysiological or optical recordings, providing powerful approaches to simultaneously monitor and perturb neural function.

  • Activity-dependent labelling of opsins can be used to reactivate neural ensembles that encode particular behaviours or experiences.

  • New anatomical techniques (such as viral-tracing methods and hydrogel-embedding methods for optical access) enable molecular and anatomical profiling of the same cells that were active in vivo, providing integrative understanding of neural circuitry.

Abstract

Modern optogenetics can be tuned to evoke activity that corresponds to naturally occurring local or global activity in timing, magnitude or individual-cell patterning. This outcome has been facilitated not only by the development of core features of optogenetics over the past 10 years (microbial-opsin variants, opsin-targeting strategies and light-targeting devices) but also by the recent integration of optogenetics with complementary technologies, spanning electrophysiology, activity imaging and anatomical methods for structural and molecular analysis. This integrated approach now supports optogenetic identification of the native, necessary and sufficient causal underpinnings of physiology and behaviour on acute or chronic timescales and across cellular, circuit-level or brain-wide spatial scales.

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Figure 1: Approaches to opsin targeting with anatomical and cell type specificity.
Figure 2: Integrating optogenetic control with in vivo electrophysiology.
Figure 3: Integrating optogenetic control with optical methods: matching naturally occurring activity patterns and linking to brain-wide projection activity.

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Acknowledgements

The authors thank the members of the Deisseroth laboratory for helpful discussions; in particular, S.J.Y. and T.J.D. for helpful discussions about fibre photometry analysis. C.K.K. is supported by a National Research Service Award (NRSA) F31 award (NIDA F31DA041795). A.A. is supported by the Walter V. and Idun Berry award, a K99 award (NIMH K99MH106649), and a NARSAD Young Investigator fellowship. K.D. is supported by the National Institutes of Health (NIH), National Science Foundation (NSF), Defense Advanced Research Projects Agency (DARPA) and the Wiegers, Grosfeld, Snyder, Yu, and Woo Foundations.

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C.K.K., A.A. and K.D. wrote the paper; C.K.K. and A.A. contributed equally to this work.

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Correspondence to Karl Deisseroth.

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Glossary

Fibre-optic patch cord

A flexible and lightweight optical fibre that is used to connect a light source (such as a laser diode or a light-emitting diode (LED)) to a fibre-optic cannula implanted on an animal, allowing light delivery to target cell populations in freely moving animals.

Kinetic opsin variants

Opsin variants that have been engineered to have slower or faster deactivation kinetics, such as the stabilized step-function opsin or 'ChETA' (E123T mutation-containing channelrhodopsin) variants, respectively.

Step-function opsins

Opsin proteins with very slow deactivation kinetics, which can thus remain activated for tens of minutes following brief light delivery and can also be switched off in a temporally precise manner with a different wavelength of light.

Red-shifted excitatory opsins

Opsin proteins such as VChR1 and C1V1 that have been discovered and/or engineered to be excited by light of longer wavelengths (that is, red-shifted), in contrast to blue light-activated channelrhodopsins, making them useful for integrating optogenetic excitation with Ca2+ imaging through blue-light-excited GCaMP sensors.

Boolean logic

An algebraic framework in which the basic operations are “OR”, “NOT” and “AND”. These logical operators have been implemented for targeting cell types defined by the presence or absence of multiple features, such as through the use of multiple recombinases and INTRSECT viruses that allow expression of genes encoding opsins in neuronal populations that express Cre NOT Flp recombinase.

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Kim, C., Adhikari, A. & Deisseroth, K. Integration of optogenetics with complementary methodologies in systems neuroscience. Nat Rev Neurosci 18, 222–235 (2017). https://doi.org/10.1038/nrn.2017.15

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