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Optimization of a GCaMP calcium indicator for neural activity imaging

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dc.contributor.author Akerboom, Jasper
dc.contributor.author Chen, Tsai-Wen
dc.contributor.author Wardill, Trevor J.
dc.contributor.author Tian, Lin
dc.contributor.author Marvin, Jonathan S.
dc.contributor.author Mutlu, Sevinc
dc.contributor.author Calderon, Nicole Carreras
dc.contributor.author Esposti, Federico
dc.contributor.author Borghuis, Bart G.
dc.contributor.author Sun, Xiaonan Richard
dc.contributor.author Gordus, Andrew
dc.contributor.author Orger, Michael B.
dc.contributor.author Portugues, Ruben
dc.contributor.author Engert, Florian
dc.contributor.author Macklin, John J.
dc.contributor.author Filosa, Alessandro
dc.contributor.author Aggarwal, Aman
dc.contributor.author Kerr, Rex A.
dc.contributor.author Takagi, Ryousuke
dc.contributor.author Kracun, Sebastian
dc.contributor.author Shigetomi, Eiji
dc.contributor.author Khakh, Baljit S.
dc.contributor.author Baier, Herwig
dc.contributor.author Lagnado, Leon
dc.contributor.author Wang, Samuel S.-H.
dc.contributor.author Bargmann, Cornelia I.
dc.contributor.author Kimmel, Bruce E.
dc.contributor.author Jayaraman, Vivek
dc.contributor.author Svoboda, Karel
dc.contributor.author Kim, Douglas S.
dc.contributor.author Schreiter, Eric R.
dc.contributor.author Looger, Loren L.
dc.date.accessioned 2012-10-12T19:50:34Z
dc.date.available 2012-10-12T19:50:34Z
dc.date.issued 2012-10-03
dc.identifier.citation Journal of Neuroscience 32 (2012): 13819-13840 en_US
dc.identifier.uri http://hdl.handle.net/1912/5448
dc.description © The Author(s), 2012. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Journal of Neuroscience 32 (2012): 13819-13840, doi:10.1523/JNEUROSCI.2601-12.2012. en_US
dc.description.abstract Genetically encoded calcium indicators (GECIs) are powerful tools for systems neuroscience. Recent efforts in protein engineering have significantly increased the performance of GECIs. The state-of-the art single-wavelength GECI, GCaMP3, has been deployed in a number of model organisms and can reliably detect three or more action potentials in short bursts in several systems in vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of in vitro assays, we have increased the dynamic range of GCaMP3 by severalfold, creating a family of “GCaMP5” sensors. We tested GCaMP5s in several systems: cultured neurons and astrocytes, mouse retina, and in vivo in Caenorhabditis chemosensory neurons, Drosophila larval neuromuscular junction and adult antennal lobe, zebrafish retina and tectum, and mouse visual cortex. Signal-to-noise ratio was improved by at least 2- to 3-fold. In the visual cortex, two GCaMP5 variants detected twice as many visual stimulus-responsive cells as GCaMP3. By combining in vivo imaging with electrophysiology we show that GCaMP5 fluorescence provides a more reliable measure of neuronal activity than its predecessor GCaMP3. GCaMP5 allows more sensitive detection of neural activity in vivo and may find widespread applications for cellular imaging in general. en_US
dc.description.sponsorship A.F. has been supported by a European Molecular Biology Organization long-term fellowship. Work in H.B.’s laboratory was funded by the National Institutes of Health (NIH) Nanomedicine Development Center “Optical Control of Biological Function,” and work in S.S.-H.W.’s laboratory was funded by NIH R01 NS045193. en_US
dc.format.mimetype application/pdf
dc.language.iso en_US en_US
dc.publisher Society for Neuroscience en_US
dc.relation.uri http://dx.doi.org/10.1523/JNEUROSCI.2601-12.2012
dc.rights.uri http://creativecommons.org/licenses/by-nc-sa/3.0/ *
dc.title Optimization of a GCaMP calcium indicator for neural activity imaging en_US
dc.type Article en_US
dc.identifier.doi 10.1523/JNEUROSCI.2601-12.2012


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