Universität zu Köln

Hochbaum, Daniel R., Zhao, Yongxin ORCID: 0000-0003-4188-5725, Farhi, Samouil L., Klapoetke, Nathan ORCID: 0000-0002-3270-1966, Werley, Christopher A., Kapoor, Vikrant, Zou, Peng ORCID: 0000-0002-9798-5242, Kralj, Joel M., Maclaurin, Dougal, Smedemark-Margulies, Niklas, Saulnier, Jessica L., Boulting, Gabriella L., Straub, Christoph, Cho, Yong Ku ORCID: 0000-0001-5621-521X, Melkonian, Michael, Wong, Gane Ka-Shu, Harrison, D. Jed, Murthy, Venkatesh N., Sabatini, Bernardo L., Boyden, Edward S., Campbell, Robert E. ORCID: 0000-0003-0604-092X and Cohen, Adam E. (2014). All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins. Nat. Methods, 11 (8). S. 825 - 834. LONDON: NATURE PUBLISHING GROUP. ISSN 1548-7105

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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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Hochbaum, Daniel R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Zhao, Yongxin UNSPECIFIED orcid.org/0000-0003-4188-5725 UNSPECIFIED Farhi, Samouil L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Klapoetke, Nathan UNSPECIFIED orcid.org/0000-0002-3270-1966 UNSPECIFIED Werley, Christopher A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kapoor, Vikrant UNSPECIFIED UNSPECIFIED UNSPECIFIED Zou, Peng UNSPECIFIED orcid.org/0000-0002-9798-5242 UNSPECIFIED Kralj, Joel M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Maclaurin, Dougal UNSPECIFIED UNSPECIFIED UNSPECIFIED Smedemark-Margulies, Niklas UNSPECIFIED UNSPECIFIED UNSPECIFIED Saulnier, Jessica L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Boulting, Gabriella L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Straub, Christoph UNSPECIFIED UNSPECIFIED UNSPECIFIED Cho, Yong Ku UNSPECIFIED orcid.org/0000-0001-5621-521X UNSPECIFIED Melkonian, Michael UNSPECIFIED UNSPECIFIED UNSPECIFIED Wong, Gane Ka-Shu UNSPECIFIED UNSPECIFIED UNSPECIFIED Harrison, D. Jed UNSPECIFIED UNSPECIFIED UNSPECIFIED Murthy, Venkatesh N. UNSPECIFIED UNSPECIFIED UNSPECIFIED Sabatini, Bernardo L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Boyden, Edward S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Campbell, Robert E. UNSPECIFIED orcid.org/0000-0003-0604-092X UNSPECIFIED Cohen, Adam E. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-433405
DOI:	10.1038/NMETH.3000
Journal or Publication Title:	Nat. Methods
Volume:	11
Number:	8
Page Range:	S. 825 - 834
Date:	2014
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	1548-7105
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PLURIPOTENT STEM-CELLS; AXON INITIAL SEGMENT; SENSITIVE FLUORESCENT PROTEIN; ACTION-POTENTIALS; PYRAMIDAL NEURONS; NEURAL ACTIVITY; VOLTAGE; EXCITABILITY; LIGHT; STIMULATION Multiple languages Biochemical Research Methods Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/43340