Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures

By Feng Zhang, Viviana Gradinaru, Antoine Adamantidis, R. Durand, Raag D. Airan, Luis de Lecea, Karl Deisseroth. 2010

718 citations recorded by the source corpus.

Elucidation of the neural substrates underlying complex animal behaviors depends on precise activity control tools, as well as compatible readout methods. Recent developments in optogenetics have addressed this need, opening up new possibilities for systems neuroscience. Interrogation of even deep neural circuits can be conducted by directly probing the necessity and sufficiency of defined circuit elements with millisecond-scale, cell type-specific optical perturbations, coupled with suitable readouts such as electrophysiology, optical circuit dynamics measures and freely moving behavior in mammals. Here we collect in detail our strategies for delivering microbial opsin genes to deep mammalian brain structures in vivo, along with protocols for integrating the resulting optical control with compatible readouts (electrophysiological, optical and behavioral). The procedures described here, from initial virus preparation to systems-level functional readout, can be completed within 4–5 weeks. Together, these methods may help in providing circuit-level insight into the dynamics underlying complex mammalian behaviors in health and disease.

Subfields: Cellular and Molecular Neuroscience, Cognitive Neuroscience

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Citations in the corpus

Adult Neurogenesis in the Mammalian Brain: Significant Answers and Significant Questions (2011, background): “New technologies, such as optogenetics (reviewed by Zhang et al., 2010), transneuronal tracers (reviewed by Callaway, 2008), and in vivo imaging, will help to address these questions.”
Neuronal homeostasis: time for a change? (2011, background): “Optogenetic methods (Gradinaru et al. 2010; Zhang et al. 2010) permit non-invasive measurement and manipulation of electrical signals on the single-cell level over timescales that potentially span the lifetime of an organism.”
The Development and Application of Optogenetics (2011, methodology): “This doublefloxed inverted open-reading-frame (DIO; reviewed in Zhang et al. 2010) strategy (Atasoy et al. 2008, Sohal et al. 2009, Tsai et al. 2009) situates the opsin gene (inhibitory or excitatory) in the inverted (meaningless) orientation, but the gene is flanked by two sets of incompatible Cre…”
Molecular and cellular approaches for diversifying and extending optogenetics. (2010, background): “…choice, virus titer, virus tropism, time of opsin expression, target cell biophysics, and local patterns of endogenous activity and modulation will all affect optogenetic inhibition efficacy and should be carefully considered for each experimental system (Cardin et al., 2010; Zhang et al., 2010).”
Permanent Genetic Access to Transiently Active Neurons via TRAP: Targeted Recombination in Active Populations (2013, background): “For instance, one common strategy to express a tool in a particular cell type and brain region is to use local injections of Cre-dependent viruses into genetically engineered mice that express Cre recombinase in a specific cell type (Zhang et al., 2010).”
Projection-specific modulation of dopamine neuron synapses by aversive and rewarding stimuli (2011, positive): “Over the last decade, important new approaches have become available to facilitate this effort ranging from genetically modified mice in which transgenes are expressed in specific cell types (Malenka, 2002) to optogenetics (Zhang et al., 2010b).”
Optical Control of Mammalian Endogenous Transcription and Epigenetic States (2013, neutral): “5b) 28 .”
Dopamine release in the basal ganglia (2011, background): “However, advances in optogenetics permit optical stimulation (or suppression) of specific cell types (Deisseroth, 2010; 2011; Zhang et al., 2010; Fenno et al., 2011; Kravitz and Kreitzer, 2011; Toettcher et al., 2011), allowing new questions about DA release to be addressed.”