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dc.contributor.authorAkerboom, Jasper
dc.contributor.authorCarreras Calderón, Nicole
dc.contributor.authorTian, Lin
dc.contributor.authorWabnig, Sebastian
dc.contributor.authorPrigge, Matthias
dc.contributor.authorTolö, Johan
dc.contributor.authorGordus, Andrew
dc.contributor.authorOrger, Michael B
dc.contributor.authorSeveri, Kristen E
dc.contributor.authorMacklin, John J
dc.contributor.authorPatel, Ronak
dc.contributor.authorPulver, Stefan R
dc.contributor.authorWardill, Trevor J
dc.contributor.authorFischer, Elisabeth
dc.contributor.authorSchüler, Christina
dc.contributor.authorChen, Tsai-Wen
dc.contributor.authorSarkisyan, Karen S
dc.contributor.authorMarvin, Jonathan S
dc.contributor.authorBargmann, Cornelia I
dc.contributor.authorKim, Douglas S
dc.contributor.authorKügler, Sebastian
dc.contributor.authorLagnado, Leon
dc.contributor.authorHegemann, Peter
dc.contributor.authorGottschalk, Alexander
dc.contributor.authorSchreiter, Eric R
dc.contributor.authorLooger, Loren L
dc.identifier.citationAkerboom , J , Carreras Calderón , N , Tian , L , Wabnig , S , Prigge , M , Tolö , J , Gordus , A , Orger , M B , Severi , K E , Macklin , J J , Patel , R , Pulver , S R , Wardill , T J , Fischer , E , Schüler , C , Chen , T-W , Sarkisyan , K S , Marvin , J S , Bargmann , C I , Kim , D S , Kügler , S , Lagnado , L , Hegemann , P , Gottschalk , A , Schreiter , E R & Looger , L L 2013 , ' Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics ' , Frontiers in Molecular Neuroscience , vol. 6 , 2 .
dc.identifier.otherPURE: 167959987
dc.identifier.otherPURE UUID: 15c02b2f-3ae9-46fc-8cb3-c9bcb09ee361
dc.identifier.otherPubMed: 23459413
dc.identifier.otherScopus: 84874185263
dc.identifier.otherORCID: /0000-0001-5170-7522/work/69463419
dc.descriptionThis work was supported by Goethe University Frankfurt; Deutsche Forschungsgemeinschaft (DFG), grant EXC115; European Union FP7 grant “EUTrigTreat and German Research Council (DFG) Grant MI 685/2-1.en
dc.description.abstractGenetically 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.
dc.relation.ispartofFrontiers in Molecular Neuroscienceen
dc.rightsCopyright © 2013 Akerboom, Carreras Calderón, Tian, Wabnig, Prigge, Tolö, Gordus, Orger, Severi, Macklin, Patel, Pulver, Wardill, Fischer, Schüler, Chen, Sarkisyan, Marvin, Bargmann, Kim, Kügler, Lagnado, Hegemann, Gottschalk, Schreiter and Looger. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.en
dc.subjectRC0321 Neuroscience. Biological psychiatry. Neuropsychiatryen
dc.titleGenetically encoded calcium indicators for multi-color neural activity imaging and combination with optogeneticsen
dc.typeJournal articleen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews.School of Psychology and Neuroscienceen
dc.description.statusPeer revieweden

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