Person: Portugues, Ruben
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Publication Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging
(Society for Neuroscience, 2012) 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.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.