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Nat Methods ; 12(9): 852-8, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26167640

RESUMO

Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca(2+) indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca(2+) detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).


Assuntos
Sinalização do Cálcio/fisiologia , Cálcio/metabolismo , Proteínas Luminescentes/metabolismo , Neurônios/citologia , Neurônios/fisiologia , Optogenética/métodos , Animais , Rastreamento de Células/métodos , Células Cultivadas , Drosophila , Luz , Proteínas Luminescentes/genética , Microscopia de Fluorescência/métodos , Engenharia de Proteínas/métodos , Ratos , Peixe-Zebra
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