Optimization of a GCaMP calcium indicator for neural activity imaging

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Date
2012-10-03Author
Akerboom, Jasper
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Chen, Tsai-Wen
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Wardill, Trevor J.
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Tian, Lin
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Marvin, Jonathan S.
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Mutlu, Sevinc
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Calderon, Nicole Carreras
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Esposti, Federico
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Borghuis, Bart G.
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Sun, Xiaonan Richard
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Gordus, Andrew
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Orger, Michael B.
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Portugues, Ruben
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Engert, Florian
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Macklin, John J.
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Filosa, Alessandro
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Aggarwal, Aman
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Kerr, Rex A.
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Takagi, Ryousuke
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Kracun, Sebastian
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Shigetomi, Eiji
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Khakh, Baljit S.
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Baier, Herwig
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Lagnado, Leon
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Wang, Samuel S.-H.
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Bargmann, Cornelia I.
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Kimmel, Bruce E.
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Jayaraman, Vivek
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Svoboda, Karel
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Kim, Douglas S.
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Schreiter, Eric R.
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Looger, Loren L.
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https://hdl.handle.net/1912/5448As published
https://doi.org/10.1523/JNEUROSCI.2601-12.2012DOI
10.1523/JNEUROSCI.2601-12.2012Abstract
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.
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© 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.
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Journal of Neuroscience 32 (2012): 13819-13840The following license files are associated with this item: