Optimization of a GCaMP calcium indicator for neural activity imaging

By Jasper Akerboom, Tsai-Wen Chen, T. Wardill, Lin Tian, J. Marvin, Sevinç Mutlu, Nicole Carreras Calderón, F. Esposti, B. Borghuis, Xiaonan R. Sun and 22 more. Journal of Neuroscience, 2012

1,194 citations recorded by the source corpus.

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.

Subfields: Biophysics, Cellular and Molecular Neuroscience, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/4c61e65dab52f90646f86140a828d16c6b785233

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Citations in the corpus

Parvalbumin-expressing interneurons linearly control olfactory bulb output (2013, neutral): “The genetically encoded calcium indicator GCaMP5G (Akerboom et al., 2012) was expressed in PV cells by injecting a Cre-dependent viral vector into the olfactory bulbs ofPV-Cremice (n = 5mice).”
Cortical feedback decorrelates olfactory bulb output in awake mice (2015, background): “In the presence of muscimol, single pulses (100 ms, 50 mA, see Experimental Procedures) failed to evoke calcium transients, suggesting that GCaMP5 activity may not report single action potentials (Akerboom et al., 2012).”
Essential role of the mushroom body in context-dependent CO₂ avoidance in Drosophila. (2013, background): “0 [21] under the control of MB drivers (Figures 2 and S2).”
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Origins of Cell-Type-Specific Olfactory Processing in the Drosophila Mushroom Body Circuit. (2017, positive): “To test this hypothesis, we expressed GCaMP5 (Akerboom et al., 2012) in all KCs under the control of OK107Gal4 (Connolly et al.”
Nanotools for Neuroscience and Brain Activity Mapping (2013, background): “niques are circumventing many of these issues).(7,79,80) Furthermore, the”
A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging (2016, background): “…the high resolution and image contrast provided by 2-photon microscopy allows extraction of fluorescence signals corresponding to the activity of individual neurons and subcellular compartments (Stosiek et al. 2003, Sato et al. 2007, Akerboom et al. 2012, Chen et al. 2013) (Petreanu et al. 2012).”