Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging
- Authors
- Akerboom, J., Chen, T.W., Wardill, T.J., Tian, L., Marvin, J.S., Mutlu, S., Calderón, N.C., Esposti, F., Borghuis, B.G., Sun, X.R., Gordus, A., Orger, M.B., Portugues, R., Engert, F., 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., Bargmann, C.I., Kimmel, B.E., Jayaraman, V., Svoboda, K., Kim, D.S., Schreiter, E.R., and Looger, L.L.
- ID
- ZDB-PUB-121012-31
- Date
- 2012
- Source
- The Journal of neuroscience : the official journal of the Society for Neuroscience 32(40): 13819-13840 (Journal)
- Registered Authors
- Baier, Herwig, Engert, Florian, Lagnado, Leon, Orger, Mike
- Keywords
- none
- MeSH Terms
-
- Animals
- Astrocytes/chemistry
- Astrocytes/ultrastructure
- Caenorhabditis elegans
- Calcium Signaling*
- Crystallography, X-Ray
- Drosophila melanogaster/growth & development
- Female
- Fluorescent Dyes/analysis
- Fluorescent Dyes/chemistry*
- Fluorometry/methods*
- Genes, Synthetic
- Genetic Vectors
- Green Fluorescent Proteins/analysis
- Green Fluorescent Proteins/chemistry*
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/isolation & purification
- HEK293 Cells/chemistry
- HEK293 Cells/ultrastructure
- Hippocampus/chemistry
- Hippocampus/cytology
- Humans
- Larva
- Lasers
- Mice
- Models, Molecular
- Mutagenesis, Site-Directed
- Neuroimaging/methods*
- Neuromuscular Junction/chemistry
- Neuromuscular Junction/ultrastructure
- Neurons/chemistry*
- Neurons/physiology
- Neurons/ultrastructure
- Neuropil/chemistry
- Neuropil/physiology
- Neuropil/ultrastructure
- Olfactory Receptor Neurons/chemistry
- Olfactory Receptor Neurons/physiology
- Olfactory Receptor Neurons/ultrastructure
- Peptides/analysis
- Peptides/chemistry*
- Peptides/genetics
- Photic Stimulation
- Protein Conformation
- Rats
- Recombinant Fusion Proteins/analysis
- Recombinant Fusion Proteins/chemistry
- Recombinant Fusion Proteins/genetics
- Retinal Bipolar Cells/chemistry
- Retinal Bipolar Cells/physiology
- Retinal Bipolar Cells/ultrastructure
- Synaptic Transmission*
- Zebrafish/growth & development
- PubMed
- 23035093 Full text @ J. Neurosci.
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.