Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging

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Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging

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Title: Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging
Author: Akerboom, J.; Chen, T.-W.; Wardill, T. J.; Tian, L.; Marvin, J. S.; Mutlu, S.; Calderon, N. C.; Esposti, F.; Borghuis, B. G.; Sun, X. R.; Gordus, A.; Orger, M. B.; Portugues, Ruben; Engert, Florian; Macklin, J. J.; Filosa, A.; Aggarwal, A.; Kerr, R. A.; Takagi, R.; Kracun, S.; Shigetomi, E.; Khakh, B. S.; Baier, H.; Lagnado, L.; Wang, S. S.- H.; Bargmann, C. I.; Kimmel, B. E.; Jayaraman, V.; Svoboda, K.; Kim, D. S.; Schreiter, E. R.; Looger, L. L.

Note: Order does not necessarily reflect citation order of authors.

Citation: Akerboom, Jasper, Tsai-Wen Chen, Trevor J. Wardill, Lin Tian, Jonathan S. Marvin, Sevinç Mutlu, Nicole Carreras Calderon, et al. 2012. Optimization of a GCaMP calcium indicator for neural activity imaging. Journal of Neuroscience 32(40): 13819-13840.
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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.
Published Version: doi:10.1523/JNEUROSCI.2601-12.2012
Other Sources: https://darchive.mblwhoilibrary.org/handle/1912/5448
Terms of Use: This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA
Citable link to this page: http://nrs.harvard.edu/urn-3:HUL.InstRepos:11315420
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