RRC ID 47678
Author Ohkura M, Sasaki T, Kobayashi C, Ikegaya Y, Nakai J.
Title An improved genetically encoded red fluorescent Ca2+ indicator for detecting optically evoked action potentials.
Journal PLoS One
Abstract Genetically encoded Ca(2+) indicators (GECIs) are powerful tools to image activities of defined cell populations. Here, we developed an improved red fluorescent GECI, termed R-CaMP1.07, by mutagenizing R-GECO1. In HeLa cell assays, R-CaMP1.07 exhibited a 1.5-2-fold greater fluorescence response compared to R-GECO1. In hippocampal pyramidal neurons, R-CaMP1.07 detected Ca(2+) transients triggered by single action potentials (APs) with a probability of 95% and a signal-to-noise ratio >7 at a frame rate of 50 Hz. The amplitudes of Ca(2+) transients linearly correlated with the number of APs. The expression of R-CaMP1.07 did not significantly alter the electrophysiological properties or synaptic activity patterns. The co-expression of R-CaMP1.07 and channelrhodpsin-2 (ChR2), a photosensitive cation channel, in pyramidal neurons demonstrated that R-CaMP1.07 was applicable for the monitoring of Ca(2+) transients in response to optically evoked APs, because the excitation light for R-CaMP1.07 hardly activated ChR2. These technical advancements provide a novel strategy for monitoring and manipulating neuronal activity with single cell resolution.
Volume 7(7)
Pages e39933
Published 2012-1-1
DOI 10.1371/journal.pone.0039933
PII PONE-D-12-12269
PMID 22808076
PMC PMC3393713
MeSH Action Potentials / physiology* Action Potentials / radiation effects Animals Animals, Newborn Calcium / analysis* Calcium / metabolism Channelrhodopsins Escherichia coli Fluorescent Dyes Gene Expression Genes, Reporter* HeLa Cells Humans Light Molecular Probes* Neurons / physiology* Neurons / radiation effects Pyramidal Cells / physiology* Pyramidal Cells / radiation effects Rats Rats, Wistar Recombinant Proteins / genetics Recombinant Proteins / metabolism Signal-To-Noise Ratio Single-Cell Analysis Transgenes
IF 2.74
Times Cited 73
DNA material R-CaMP1.07 (RDB14608)