FIGURE

Fig. 1

ID

ZDB-FIG-170829-17

Publication

Förster et al., 2017 - An optogenetic toolbox for unbiased discovery of functionally connected cells in neural circuits

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Fig. 1

Optobow allows all-optical mapping of connected neurons. a Design of the Optobow-c construct. mCerulean and tdTomato are both membrane-targeted (CAAX motif). b Sketch for Optobow-c expression in the zebrafish optic tectum. Different cell types are indicated by their arborization patterns in the tectal neuropil. Addition of Cre by transient injections results in random expression of either ChrimsonR (labeled by tdTomato, magenta) or GCaMP6f (green). Unrecombined cells are labeled by mCerulean (blue). Upon photostimulation of a single ChrimsonR cell (dashed orange outline) connected cells will be highlighted by a rise in GCaMP fluorescence (dark green cell). c In vivo tectal expression of Optobow-c. Scale bar, 20 µm. d Microscope setup for all-optical connectivity mapping. Two independent infrared-pulsed lasers are used to photostimulate ChrimsonR cells (magenta) and to image GCaMP fluorescence (green), respectively. A spatial light modulator (SLM) allows precisely targeted stimulation of single cells. See methods for abbreviations. e Dorsal view of tectal Optobow-c expression. Single fluorescent channels show a cluster of three ChrimsonR- (left) and 18 GCaMP6f-expressing cells (middle). Photostimulation was either confined to a single ChrimsonR cell (Stim1, dashed orange outline) or to a control region of equal size (Stim2). White dashed line marks skin. Dotted rectangles show regions of calcium imaging during photostimulation. A close-up showing a single confocal plane (z = 2.5 µm) of the photostimulated region (Stim1) is shown in the merge on the right. Despite the close proximity of the ChrimsonR and the GCaMP cell#2, exclusive expression of either component allows restricted stimulation of the ChrimsonR-expressing cell. Scale bar, 15 µm. f Calcium transients acquired at 10 fps shown as Z-scores, obtained simultaneously from the regions indicated in e. Photostimulation events are highlighted by dashed lines with stimulation lengths indicated below. Off-target stimulation did not result in significant GCaMP activity (Stim2)

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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