Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering.

Autor:	Chen Y; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA., Jang H; Buck Institute for Research on Aging, Novato, CA 94945, USA., Spratt PWE; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA., Kosar S; Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA., Taylor DE; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA., Essner RA; Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA., Bai L; Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA., Leib DE; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA., Kuo TW; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA., Lin YC; Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA., Patel M; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA., Subkhangulova A; Buck Institute for Research on Aging, Novato, CA 94945, USA., Kato S; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA; Department of Neurology, University of California, San Francisco, San Francisco, CA 94158, USA., Feinberg EH; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA; Department of Anatomy, University of California, San Francisco, San Francisco, CA 94158, USA., Bender KJ; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA; Department of Neurology, University of California, San Francisco, San Francisco, CA 94158, USA., Knight ZA; Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: zachary.knight@ucsf.edu., Garrison JL; Buck Institute for Research on Aging, Novato, CA 94945, USA; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: jgarrison@buckinstitute.org.
Jazyk:	angličtina
Zdroj:	Neuron [Neuron] 2020 Aug 05; Vol. 107 (3), pp. 454-469.e6. Date of Electronic Publication: 2020 Jun 22.
DOI:	10.1016/j.neuron.2020.05.005
Abstrakt:	Neuroscience relies on techniques for imaging the structure and dynamics of neural circuits, but the cell bodies of individual neurons are often obscured by overlapping fluorescence from axons and dendrites in surrounding neuropil. Here, we describe two strategies for using the ribosome to restrict the expression of fluorescent proteins to the neuronal soma. We show first that a ribosome-tethered nanobody can be used to trap GFP in the cell body, thereby enabling direct visualization of previously undetectable GFP fluorescence. We then design a ribosome-tethered GCaMP for imaging calcium dynamics. We show that this reporter faithfully tracks somatic calcium dynamics in the mouse brain while eliminating cross-talk between neurons caused by contaminating neuropil. In worms, this reporter enables whole-brain imaging with faster kinetics and brighter fluorescence than commonly used nuclear GCaMPs. These two approaches provide a general way to enhance the specificity of imaging in neurobiology. Competing Interests: Declaration of Interests The authors declare no competing interests. (Copyright © 2020. Published by Elsevier Inc.)
Databáze:	MEDLINE
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