All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins
Name
Boyden_All-optical electrophysiology.pdf
Size
2.09 MB
Format
Adobe PDF
Checksum (MD5)
703d6daf5e5efa29a8fdeeff921f13a7
Author(s)
Hochbaum, Daniel R
Zhao, Yongxin
Farhi, Samouil L
Werley, Christopher A
Kapoor, Vikrant
Zou, Peng
Kralj, Joel M
Maclaurin, Dougal
Smedemark-Margulies, Niklas
Saulnier, Jessica L
Date Issued
June 2014
Journal
Nature Methods
Publisher
Nature Publishing Group
Citation
Hochbaum, Daniel R; Zhao, Yongxin; Farhi, Samouil L; Klapoetke, Nathan; Werley, Christopher A; Kapoor, Vikrant; Zou, Peng et al. “All-Optical Electrophysiology in Mammalian Neurons Using Engineered Microbial Rhodopsins.” Nature Methods 11, no. 8 (June 2014): 825–833 © 2014 Rights Managed by Nature Publishing Group
Version
Author's final manuscript
Abstract
All-optical electrophysiology—spatially resolved simultaneous optical perturbation and measurement of membrane voltage—would open new vistas in neuroscience research. We evolved two archaerhodopsin-based voltage indicators, QuasAr1 and QuasAr2, which show improved brightness and voltage sensitivity, have microsecond response times and produce no photocurrent. We engineered a channelrhodopsin actuator, CheRiff, which shows high light sensitivity and rapid kinetics and is spectrally orthogonal to the QuasArs. A coexpression vector, Optopatch, enabled cross-talk–free genetically targeted all-optical electrophysiology. In cultured rat neurons, we combined Optopatch with patterned optical excitation to probe back-propagating action potentials (APs) in dendritic spines, synaptic transmission, subcellular microsecond-timescale details of AP propagation, and simultaneous firing of many neurons in a network. Optopatch measurements revealed homeostatic tuning of intrinsic excitability in human stem cell–derived neurons. In rat brain slices, Optopatch induced and reported APs and subthreshold events with high signal-to-noise ratios. The Optopatch platform enables high-throughput, spatially resolved electrophysiology without the use of conventional electrodes.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Media Laboratory
McGovern Institute for Brain Research at MIT
Terms of Use
Creative Commons Attribution-Noncommercial-Share Alike
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/nmeth.3000
