Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging

Circuit mapping requires knowledge of both structural and functional connectivity between cells. While 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 (pa-GECIs) that combines attributes of high-contrast photo-labeling with high-sensitivity Ca(2+) detection in a single-color, protein-sensor. We demonstrate the utility of pa-GECIs in cultured neurons and in vivo in Drosophila and zebrafish larvae. We show how single cells can be selected out of dense populations for Golgi-like visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is readily transferrable to other sensors based on circularly permutated GFP (cpGFP)