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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 Bhailal
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.date.accessioned2014-02-21T20:28:18Z
dc.date.issued2013
dc.identifier.citationAkerboom, Jasper, Nicole Carreras Calderón, Lin Tian, Sebastian Wabnig, Matthias Prigge, Johan Tolö, Andrew Gordus, and et al. 2013. Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics. Frontiers in Molecular Neuroscience 6:2.en_US
dc.identifier.issn1662-5099en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:11732109
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.en_US
dc.description.sponsorshipMolecular and Cellular Biologyen_US
dc.language.isoen_USen_US
dc.publisherFrontiers Media S.A.en_US
dc.relation.isversionofdoi:10.3389/fnmol.2013.00002en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3586699/pdf/en_US
dash.licenseLAA
dc.subjectcalcium imagingen_US
dc.subjectgenetically encoded calcium indicatoren_US
dc.subjectmulti-color imagingen_US
dc.subjectprotein engineeringen_US
dc.subjectoptogeneticsen_US
dc.titleGenetically Encoded Calcium Indicators for Ulti-Color Neural Activity Imaging and Combination With Optogeneticsen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalFrontiers in Molecular Neuroscienceen_US
dash.depositing.authorPatel, Ronak Bhailal
dc.date.available2014-02-21T20:28:18Z
dc.identifier.doi10.3389/fnmol.2013.00002*
workflow.legacycommentsFLAG8 HMSen_US
dash.authorsorderedfalse
dash.contributor.affiliatedPatel, Ronak Bhailal


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