Optimization of a GCaMP calcium indicator for neural activity imaging

Jasper Akerboom, Tsai Wen Chen, Trevor J. Wardill, Lin Tian, Jonathan S. Marvin, Sevinç Mutlu, Nicole Carreras Calderón, Federico Esposti, Bart G. Borghuis, Xiaonan Richard Sun, Andrew Gordus, Michael B. Orger, Ruben Portugues, Florian Engert, John J. Macklin, Alessandro Filosa, Aman Aggarwal, Rex A. Kerr, Ryousuke Takagi, Sebastian Kracun & 12 others Eiji Shigetomi, Baljit S. Khakh, Herwig Baier, Leon Lagnado, Samuel S H Wang, Cornelia I. Bargmann, Bruce E. Kimmel, Vivek Jayaraman, Karel Svoboda, Douglas S. Kim, Eric R. Schreiter, Loren L. Looger

Research output: Contribution to journalArticle

603 Citations (Scopus)

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.GCaMP5allows more sensitive detection of neural activity in vivo and may find widespread applications for cellular imaging in general.

Original languageEnglish (US)
Pages (from-to)13819-13840
Number of pages22
JournalJournal of Neuroscience
Volume32
Issue number40
DOIs
StatePublished - Oct 3 2012

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Visual Cortex
Calcium
Retina
Caenorhabditis
Neurons
Protein Engineering
Neuromuscular Junction
Electrophysiology
Signal-To-Noise Ratio
Zebrafish
Neurosciences
Mutagenesis
Astrocytes
Action Potentials
Drosophila
Fluorescence
Proteins
In Vitro Techniques

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Akerboom, J., Chen, T. W., Wardill, T. J., Tian, L., Marvin, J. S., Mutlu, S., ... Looger, L. L. (2012). Optimization of a GCaMP calcium indicator for neural activity imaging. Journal of Neuroscience, 32(40), 13819-13840. https://doi.org/10.1523/JNEUROSCI.2601-12.2012

Optimization of a GCaMP calcium indicator for neural activity imaging. / Akerboom, Jasper; Chen, Tsai Wen; Wardill, Trevor J.; Tian, Lin; Marvin, Jonathan S.; Mutlu, Sevinç; Calderón, 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.

In: Journal of Neuroscience, Vol. 32, No. 40, 03.10.2012, p. 13819-13840.

Research output: Contribution to journalArticle

Akerboom, J, Chen, TW, Wardill, TJ, Tian, L, Marvin, JS, Mutlu, S, Calderón, NC, Esposti, F, Borghuis, BG, Sun, XR, Gordus, A, Orger, MB, Portugues, R, Engert, F, Macklin, JJ, Filosa, A, Aggarwal, A, Kerr, RA, Takagi, R, Kracun, S, Shigetomi, E, Khakh, BS, Baier, H, Lagnado, L, Wang, SSH, Bargmann, CI, Kimmel, BE, Jayaraman, V, Svoboda, K, Kim, DS, Schreiter, ER & Looger, LL 2012, 'Optimization of a GCaMP calcium indicator for neural activity imaging', Journal of Neuroscience, vol. 32, no. 40, pp. 13819-13840. https://doi.org/10.1523/JNEUROSCI.2601-12.2012
Akerboom, Jasper ; Chen, Tsai Wen ; Wardill, Trevor J. ; Tian, Lin ; Marvin, Jonathan S. ; Mutlu, Sevinç ; Calderón, 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. / Optimization of a GCaMP calcium indicator for neural activity imaging. In: Journal of Neuroscience. 2012 ; Vol. 32, No. 40. pp. 13819-13840.
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AU - Marvin, Jonathan S.

AU - Mutlu, Sevinç

AU - Calderón, Nicole Carreras

AU - Esposti, Federico

AU - Borghuis, Bart G.

AU - Sun, Xiaonan Richard

AU - Gordus, Andrew

AU - Orger, Michael B.

AU - Portugues, Ruben

AU - Engert, Florian

AU - Macklin, John J.

AU - Filosa, Alessandro

AU - Aggarwal, Aman

AU - Kerr, Rex A.

AU - Takagi, Ryousuke

AU - Kracun, Sebastian

AU - Shigetomi, Eiji

AU - Khakh, Baljit S.

AU - Baier, Herwig

AU - Lagnado, Leon

AU - Wang, Samuel S H

AU - Bargmann, Cornelia I.

AU - Kimmel, Bruce E.

AU - Jayaraman, Vivek

AU - Svoboda, Karel

AU - Kim, Douglas S.

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