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PDBsum entry 3sg6
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Fluorescent protein
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PDB id
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3sg6
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References listed in PDB file
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Key reference
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Title
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Optimization of a gcamp calcium indicator for neural activity imaging.
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Authors
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J.Akerboom,
T.W.Chen,
T.J.Wardill,
L.Tian,
J.S.Marvin,
S.Mutlu,
N.C.Calderón,
F.Esposti,
B.G.Borghuis,
X.R.Sun,
A.Gordus,
M.B.Orger,
R.Portugues,
F.Engert,
J.J.Macklin,
A.Filosa,
A.Aggarwal,
R.A.Kerr,
R.Takagi,
S.Kracun,
E.Shigetomi,
B.S.Khakh,
H.Baier,
L.Lagnado,
S.S.Wang,
C.I.Bargmann,
B.E.Kimmel,
V.Jayaraman,
K.Svoboda,
D.S.Kim,
E.R.Schreiter,
L.L.Looger.
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Ref.
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J Neurosci, 2012,
32,
13819-13840.
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PubMed id
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Abstract
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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.
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