PMID- 23459413
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20130306
LR  - 20220309
IS  - 1662-5099 (Print)
IS  - 1662-5099 (Electronic)
IS  - 1662-5099 (Linking)
VI  - 6
DP  - 2013
TI  - Genetically encoded calcium indicators for multi-color neural activity imaging 
      and combination with optogenetics.
PG  - 2
LID - 10.3389/fnmol.2013.00002 [doi]
LID - 2
AB  - Genetically encoded calcium indicators (GECIs) are powerful tools for systems 
      neuroscience. Here we describe red, single-wavelength GECIs, "RCaMPs," engineered 
      from circular permutation of the thermostable red fluorescent protein mRuby. 
      High-resolution crystal structures of mRuby, the red sensor RCaMP, and the 
      recently published red GECI R-GECO1 give insight into the chromophore 
      environments of the Ca(2+)-bound state of the sensors and the engineered protein 
      domain interfaces of the different indicators. We characterized the biophysical 
      properties and performance of RCaMP sensors in vitro and in vivo in 
      Caenorhabditis elegans, Drosophila larvae, and larval zebrafish. Further, we 
      demonstrate 2-color calcium imaging both within the same cell (registering 
      mitochondrial and somatic [Ca(2+)]) and between two populations of cells: neurons 
      and astrocytes. Finally, we perform integrated optogenetics experiments, wherein 
      neural activation via channelrhodopsin-2 (ChR2) or a red-shifted variant, and 
      activity imaging via RCaMP or GCaMP, are conducted simultaneously, with the 
      ChR2/RCaMP pair providing independently addressable spectral channels. Using this 
      paradigm, we measure calcium responses of naturalistic and ChR2-evoked muscle 
      contractions in vivo in crawling C. elegans. We systematically compare the RCaMP 
      sensors to R-GECO1, in terms of action potential-evoked fluorescence increases in 
      neurons, photobleaching, and photoswitching. R-GECO1 displays higher Ca(2+) 
      affinity and larger dynamic range than RCaMP, but exhibits significant 
      photoactivation with blue and green light, suggesting that integrated 
      channelrhodopsin-based optogenetics using R-GECO1 may be subject to artifact. 
      Finally, we create and test blue, cyan, and yellow variants engineered from GCaMP 
      by rational design. This engineered set of chromatic variants facilitates new 
      experiments in functional imaging and optogenetics.
FAU - Akerboom, Jasper
AU  - Akerboom J
AD  - Janelia Farm Research Campus, Howard Hughes Medical Institute Ashburn, VA, USA.
FAU - Carreras Calderon, Nicole
AU  - Carreras Calderon N
FAU - Tian, Lin
AU  - Tian L
FAU - Wabnig, Sebastian
AU  - Wabnig S
FAU - Prigge, Matthias
AU  - Prigge M
FAU - Tolo, Johan
AU  - Tolo J
FAU - Gordus, Andrew
AU  - Gordus A
FAU - Orger, Michael B
AU  - Orger MB
FAU - Severi, Kristen E
AU  - Severi KE
FAU - Macklin, John J
AU  - Macklin JJ
FAU - Patel, Ronak
AU  - Patel R
FAU - Pulver, Stefan R
AU  - Pulver SR
FAU - Wardill, Trevor J
AU  - Wardill TJ
FAU - Fischer, Elisabeth
AU  - Fischer E
FAU - Schuler, Christina
AU  - Schuler C
FAU - Chen, Tsai-Wen
AU  - Chen TW
FAU - Sarkisyan, Karen S
AU  - Sarkisyan KS
FAU - Marvin, Jonathan S
AU  - Marvin JS
FAU - Bargmann, Cornelia I
AU  - Bargmann CI
FAU - Kim, Douglas S
AU  - Kim DS
FAU - Kugler, Sebastian
AU  - Kugler S
FAU - Lagnado, Leon
AU  - Lagnado L
FAU - Hegemann, Peter
AU  - Hegemann P
FAU - Gottschalk, Alexander
AU  - Gottschalk A
FAU - Schreiter, Eric R
AU  - Schreiter ER
FAU - Looger, Loren L
AU  - Looger LL
LA  - eng
GR  - MC_U105178794/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
DEP - 20130304
PL  - Switzerland
TA  - Front Mol Neurosci
JT  - Frontiers in molecular neuroscience
JID - 101477914
PMC - PMC3586699
OTO - NOTNLM
OT  - calcium imaging
OT  - genetically encoded calcium indicator
OT  - multi-color imaging
OT  - optogenetics
OT  - protein engineering
EDAT- 2013/03/06 06:00
MHDA- 2013/03/06 06:01
PMCR- 2013/01/01
CRDT- 2013/03/06 06:00
PHST- 2013/01/04 00:00 [received]
PHST- 2013/02/11 00:00 [accepted]
PHST- 2013/03/06 06:00 [entrez]
PHST- 2013/03/06 06:00 [pubmed]
PHST- 2013/03/06 06:01 [medline]
PHST- 2013/01/01 00:00 [pmc-release]
AID - 10.3389/fnmol.2013.00002 [doi]
PST - epublish
SO  - Front Mol Neurosci. 2013 Mar 4;6:2. doi: 10.3389/fnmol.2013.00002. eCollection 
      2013.