1. Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging
- Author
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Eiji Shigetomi, Sevinç Mutlu, Andrew Gordus, Florian Engert, Loren L. Looger, John J. Macklin, Eric R. Schreiter, Aman Aggarwal, Nicole Carreras Calderón, Bruce E. Kimmel, Samuel S.-H. Wang, Douglas S. Kim, Herwig Baier, Leon Lagnado, Sebastian Kracun, Karel Svoboda, Baljit S. Khakh, Xiaonan Richard Sun, Trevor J. Wardill, Ruben Portugues, Lin Tian, Cornelia I. Bargmann, Vivek Jayaraman, Federico Esposti, Bart G. Borghuis, Jonathan S. Marvin, Alessandro Filosa, Tsai Wen Chen, Ryousuke Takagi, Michael B. Orger, Jasper Akerboom, Rex Kerr, Akerboom, J, Chen, Tw, Wardill, Tj, Tian, L, Marvin, J, Mutlu, S, Calderon, Nc, Esposti, Federico, 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, B, Baier, H, Lagnado, L, Wang, Ssh, Bargmann, Ci, Kimmel, Be, Jayaraman, V, Svoboda, K, Kim, D, Schreiter, Er, and Looger, Ll
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Models, Molecular ,Retinal Bipolar Cells ,Neuropil ,Protein Conformation ,Recombinant Fusion Proteins ,Genetic Vectors ,Green Fluorescent Proteins ,Neuromuscular Junction ,Neuroimaging ,Biology ,Crystallography, X-Ray ,Hippocampus ,Synaptic Transmission ,Olfactory Receptor Neurons ,Article ,Mice ,In vivo ,Genes, Synthetic ,medicine ,Animals ,Humans ,Premovement neuronal activity ,Fluorometry ,Calcium Signaling ,Caenorhabditis elegans ,Zebrafish ,Fluorescent Dyes ,Neurons ,Systems neuroscience ,Lasers ,General Neuroscience ,Rats ,Electrophysiology ,Drosophila melanogaster ,HEK293 Cells ,Visual cortex ,medicine.anatomical_structure ,Astrocytes ,Larva ,GCaMP ,Mutagenesis, Site-Directed ,Female ,Peptides ,Tectum ,Neuroscience ,Photic Stimulation ,Preclinical imaging - 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 systemsin vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery ofin vitroassays, 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, andin vivoinCaenorhabditischemosensory neurons,Drosophilalarval 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 combiningin vivoimaging 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 activityin vivoand may find widespread applications for cellular imaging in general.
- Published
- 2012
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