Figure 6—figure supplement 1.
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- ZDB-FIG-240603-105
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- Spikol et al., 2024 - Genetically defined nucleus incertus neurons differ in connectivity and function
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Increased calcium signaling in gsc2 neurons upon optogenetic activation of the dHb. Calcium transients were imaged at 2.6 Hz before, during, and after illumination with 561 nm light in 7 dpf larvae. (A) Drawings depicting imaging of calcium transients and optogenetic activation using confocal microscopy. (B-C’) Representative maximum intensity projections of GCaMP7a fluorescence in (B) dHb and (B’) gsc2 neurons of the same larva, or (C) dHb and (C’) rln3a NI neurons of the same larva. Anterior to the top. Scale bar, 100 μm. (D-E’’) Tg(gsc2:QF2)c721 or (F-H’’) Tg(rln3a:QF2, he1.1:YFP)c836 driver lines in (D-H) TgBAC(gng8:GAL4FF)c426; Tg(UAS:GCaMP7a)zf415; Tg(QUAS:GCaMP7a)c594 larvae (D, E, F, G, H) with or (D’, E’, F’, G’, H’) without Tg(UAS:ReaChR-RFP)jf50. The average change in GCaMP7a signaling (%ΔF/F) is shown for (D, D’, F, F’) the dorsal habenulae, (E, E’) gsc2 neurons, (G, G’) rln3a NI neurons, and (H, H’) rln3a PAG neurons. Shading indicates standard deviation. Gaps at light onset and offset are due to latency in switching the laser configuration. (D’’, E’’, F’’, G’’, H’’) Average Fpost/Fpre is shown for (D’’, F’’) the dHb, (E’’) gsc2 neurons, (G’’) rln3a NI neurons, and (H’’) rln3a PAG neurons of ReaChR+ and ReaChR- larvae. Fpost is the area under the curve for 15 frames (5.8 s) during 561 nm illumination and Fpre is the area under the curve for 15 frames (5.8 s) preceding 561 nm illumination. (D’’, E’’, F’’, G’’, H’’) Black bars indicate mean ratios: (D’’) 0.75±0.15, n=6 ReaChR- larvae, 2.95±0.41, n=5 ReaChR+ larvae, ***p=0.0004. (E’’) 1.07±0.15, n=6 ReaChR- larvae, 1.86±0.17, n=5 ReaChR+ larvae, **P=0.0073. (F’’) 1.22±0.29, n=5 ReaChR- larvae, 11.08±6.54, n=5 ReaChR+ larvae, *p=0.032. (G’’) 1.82±0.32, n=5 ReaChR- larvae, 1.97±0.59, n=5 ReaChR+ larvae, = 0.83. (H’’) 2.13±0.27, n=5 ReaChR- larvae, 1.83±0.27, n=5 ReaChR+ larvae, p=0.45. Extended y-axis in F’’ to display higher values. |