Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators.
Genetically encoded calcium indicators (GECIs) can be used to image activity in defined neuronal populations. However, current GECIs produce inferior signals compared to synthetic indicators and recording electrodes, precluding detection of low firing rates. We developed a single-wavelength GCaMP2-based GECI (GCaMP3), with increased baseline fluorescence (3-fold), increased dynamic range (3-fold) and higher affinity for calcium (1.3-fold). We detected GCaMP3 fluorescence changes triggered by single action potentials in pyramidal cell dendrites, with signal-to-noise ratio and photostability substantially better than those of GCaMP2, D3cpVenus and TN-XXL. In Caenorhabditis elegans chemosensory neurons and the Drosophila melanogaster antennal lobe, sensory stimulation-evoked fluorescence responses were significantly enhanced with GCaMP3 (4-6-fold). In somatosensory and motor cortical neurons in the intact mouse, GCaMP3 detected calcium transients with amplitudes linearly dependent on action potential number. Long-term imaging in the motor cortex of behaving mice revealed large fluorescence changes in imaged neurons over months.
Tian
Lin
L
Howard Hughes Medical Institute, Ashburn, Virginia, USA.
Hires
S Andrew
SA
Mao
Tianyi
T
Huber
Daniel
D
Chiappe
M Eugenia
ME
Chalasani
Sreekanth H
SH
Petreanu
Leopoldo
L
Akerboom
Jasper
J
McKinney
Sean A
SA
Schreiter
Eric R
ER
Bargmann
Cornelia I
CI
Jayaraman
Vivek
V
Svoboda
Karel
K
Looger
Loren L
LL
D000818:Animals; D001921:Brain / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000166:cytology* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D004331:Drosophila melanogaster / Q000166:cytology* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D051379:Mice; D009474:Neurons / Q000378:metabolism*
D002118:Calcium