Details

Autor(en) / Beteiligte

• Akerboom, Jasper
• Chen, Tsai-Wen
• Wardill, Trevor J
• Tian, Lin
• Marvin, Jonathan S
• Mutlu, Sevinç
• Calderón, Nicole Carreras
• Esposti, Federico
• Borghuis, Bart G
• Sun, Xiaonan Richard
• Gordus, Andrew
• Orger, Michael B
• Portugues, Ruben
• Engert, Florian
• Macklin, John J
• Filosa, Alessandro
• Aggarwal, Aman
• Kerr, Rex A
• Takagi, Ryousuke
• Kracun, Sebastian
• Shigetomi, Eiji
• Khakh, Baljit S
• Baier, Herwig
• Lagnado, Leon
• Wang, Samuel S-H
• Bargmann, Cornelia I
• Kimmel, Bruce E
• Jayaraman, Vivek
• Svoboda, Karel
• Kim, Douglas S
• Schreiter, Eric R
• Looger, Loren L

Titel

Optimization of a GCaMP calcium indicator for neural activity imaging

Ist Teil von

• The Journal of neuroscience, 2012-10, Vol.32 (40), p.13819-13840

Ort / Verlag

United States: Society for Neuroscience

Erscheinungsjahr

2012

Link zum Volltext

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• 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.