Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging

被引：887

作者：

Akerboom, Jasper ^{[1
]}

Chen, Tsai-Wen ^{[1
]}

Wardill, Trevor J. ^{[1
]}

Tian, Lin ^{[1
]}

Marvin, Jonathan S. ^{[1
]}

Mutlu, Sevinc ^{[1
,2
]}

Calderon, Nicole Carreras ^{[1
,3
,4
]}

Esposti, Federico ^{[3
]}

Borghuis, Bart G. ^{[1
,5
]}

Sun, Xiaonan Richard ^{[6
,7
]}

Gordus, Andrew ^{[8
]}

Orger, Michael B. ^{[2
,9
]}

Portugues, Ruben ^{[9
]}

Engert, Florian ^{[9
]}

Macklin, John J. ^{[1
]}

Filosa, Alessandro ^{[10
,11
,12
]}

Aggarwal, Aman ^{[1
,13
]}

Kerr, Rex A. ^{[1
]}

Takagi, Ryousuke ^{[14
]}

Kracun, Sebastian ^{[14
]}

Shigetomi, Eiji ^{[14
]}

Khakh, Baljit S. ^{[14
]}

Baier, Herwig ^{[10
,11
,12
]}

Lagnado, Leon ^{[3
]}

Wang, Samuel S. -H. ^{[6
,7
]}

Bargmann, Cornelia I. ^{[8
]}

Kimmel, Bruce E. ^{[1
]}

Jayaraman, Vivek ^{[1
]}

Svoboda, Karel ^{[1
]}

Kim, Douglas S. ^{[1
]}

Schreiter, Eric R. ^{[1
,4
]}

Looger, Loren L. ^{[1
]}

机构：

[1] Howard Hughes Med Inst, Ashburn, VA 20147 USA

[2] Champalimaud Ctr Unknown, Champalimaud Neurosci Programme, P-1400038 Lisbon, Portugal

[3] MRC, Mol Biol Lab, Cambridge CB2 0QH, England

[4] Univ Puerto Rico Rio Piedras, Dept Chem, San Juan, PR 00931 USA

[5] Yale Univ, Sch Med, Dept Ophthalmol & Visual Sci, New Haven, CT 06511 USA

[6] Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA

[7] Princeton Univ, Princeton Neurosci Inst, Princeton, NJ 08544 USA

[8] Rockefeller Univ, Howard Hughes Med Inst, Lab Neural Circuits & Behav, New York, NY 10065 USA

[9] Harvard Univ, Ctr Brain Sci, Dept Mol & Cellular Biol, Cambridge, MA 02138 USA

[10] Univ Calif San Francisco, Dept Physiol, Program Neurosci, San Francisco, CA 94158 USA

[11] Univ Calif San Francisco, Dept Physiol, Genet Program, San Francisco, CA 94158 USA

[12] Univ Calif San Francisco, Dept Physiol, Program Dev Biol, San Francisco, CA 94158 USA

[13] Tata Inst Fundamental Res, Natl Ctr Biol Sci, Bengaluru 560065, India

[14] Univ Calif Los Angeles, Dept Physiol, Los Angeles, CA 90095 USA

来源：

JOURNAL OF NEUROSCIENCE | 2012年 / 32卷 / 40期

基金：

美国国家卫生研究院;

关键词：

PRIMARY VISUAL-CORTEX; IN-VIVO; CELLULAR RESOLUTION; NETWORK ACTIVITY; CA2+ INDICATORS; GENE-EXPRESSION; OPTICAL SENSOR; BARREL CORTEX; MOTOR-NEURONS; PROTEIN;

D O I：

10.1523/JNEUROSCI.2601-12.2012

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

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.

引用

页码：13819 / 13840

页数：22

共 89 条

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