Optimization of a GCaMP calcium indicator for neural activity imaging

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Akerboom, Jasper
Chen, Tsai-Wen
Wardill, Trevor J.
Tian, Lin
Marvin, Jonathan S.
Mutlu, Sevinc
Calderon, Nicole Carreras
Esposti, Federico
Borghuis, Bart G.
Sun, Xiaonan Richard
Gordus, Andrew
Orger, Michael B.
Portugues, Ruben
Engert, Florian
Macklin, John J.
Filosa, Alessandro
Aggarwal, Aman
Kerr, Rex A.
Takagi, Ryousuke
Kracun, Sebastian
Shigetomi, Eiji
Khakh, Baljit S.
Baier, Herwig
Lagnado, Leon
Wang, Samuel S.-H.
Bargmann, Cornelia I.
Kimmel, Bruce E.
Jayaraman, Vivek
Svoboda, Karel
Kim, Douglas S.
Schreiter, Eric R.
Looger, Loren L.
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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.
© 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): 13819-13840
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