GCaMP
GCaMP er en genetisk kodet calciumindikator oprindeligt udviklet i 2001 af Junichi Nakai.[1] Det er en syntetisk fusion af grønt fluorescerende protein (GFP), calmodulin (CaM) og M13, en peptidsekvens fra myosin letkæde kinase.[2] Når den er bundet til Ca2+, fluorescerer GCaMP grønt med en topexcitationsbølgelængde på 480 nm og en peak emission bølgelængde på 510 nm.[3] Det bruges i biologisk forskning til at måle intracellulære Ca2+ niveauer både in vitro og in vivo ved hjælp af viralt transficeret eller transgene celle- og dyrelinjer.[4] Den genetiske sekvens, der koder for GCaMP, kan indsættes under kontrol af promotorer eksklusivt for visse celletyper, hvilket muliggør celletypespecifik ekspression af GCaMP. Da Ca2+ er en second messenger, der bidrager til mange cellulære mekanismer og signalveje, tillader GCaMP forskere at kvantificere aktiviteten af Ca2+-baserede mekanismer og studere rollen af Ca2+ ioner i biologiske processer af interesse.
Referencer
- ^ Nakai J, Ohkura M, Imoto K. A high signal-to-noise Ca(2+) probe composed of a single green fluorescent protein. Nat Biotechnol. 2001 Feb;19(2):137-41. doi: 10.1038/84397. PMID 11175727.
- ^ Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, Kim DS. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. PMID 23868258; PMCID: PMC3777791.
- ^ Deciphering the molecular mechanism responsible for GCaMP6m's Ca2+-dependent change in fluorescence
- ^ Whitaker M. Genetically encoded probes for measurement of intracellular calcium. Methods Cell Biol. 2010;99:153-82. doi: 10.1016/B978-0-12-374841-6.00006-2. PMID 21035686; PMCID: PMC3292878.
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