PMC

(a) GCaMP6 response to water intake in distinct groups of PAM neurons driven by DDC-GAL4 in thirsty (left panel) or water-sated (right panel) flies, respectively. Scale bar, 20 μm. (b,c) Time-lapse recordings of GCaMP6 responses (b) in thirsty (left panel) or water-sated (right panel) flies; the average responses (c) to water intake in distinct groups of PAM neurons driven by DDC-GAL4. Each value represents mean±s.e.m. (N=8 for thirsty flies; N=11 for water-sated flies). NS, not significant (P>0.05). *P<0.05; t-test. (d) GCaMP6 response to water intake in PAM-β′1 neurons driven by VT8167-GAL4 in thirsty (left panel) or water-sated (right panel) flies, respectively. Scale bar, 20 μm. (e,f) Time-lapse recordings of GCaMP6 responses (e) in thirsty (left panel) or water-sated (right panel) flies; the average responses (f) to water intake in PAM-β′1 neurons driven by VT8167-GAL4. Each value represents mean±s.e.m. (N=8 for thirsty flies; N=10 for water-sated flies). *P<0.05; t-test.

A. Schematic of the approach for assessing changes in the similarity of cue-evoked InsCtx activity patterns to ongoing activity patterns during Thirsty and Quenched states. See for further details.
B. Top: example session with InsCtx neuronal responses to the water cue and subsequent water consumption (‘cue + consumption’, change from baseline activity). Activity was sorted by the difference in ongoing activity between Thirsty (T) and Quenched (Q) states (ΔState, right, values: −0.3 to 1.4). Bottom: average licking.
C. Correlation between InsCtx neurons’ ‘cue + consumption’ evoked responses and their changes in ongoing activity between Thirsty and Quenched states (ΔState), for the example in B.
D. Left: example pattern similarity of InsCtx peri-cue population activity in the Thirsty state to ongoing activity in the Thirsty vs. Quenched states. Mean±s.e.m. across 45 presentations of each cue. Right: mean±s.e.m across 9 FOVs from 7 mice. Gray shaded areas: cue period.
E. Pattern similarity of InsCtx peri-cue population activity during gradual quenching to ongoing activity in the Thirsty vs. Quenched states. Gray shaded areas: cue period.
F. Population activity along the Large Pupil vs. Small Pupil axis (arousal), in comparison with the Thirsty vs. Quenched axis (physiological state, see also D, right). Left: these axes are approximately orthogonal (mean angle: 97±8°). Right: pattern similarity of InsCtx peri-cue population activity during a thirsty state to ongoing activity along the Large Pupil vs. Small Pupil axis. Mean±s.e.m across 9 FOVs from 7 mice. Values of 1 and 0 correspond to mean ongoing activity patterns in epochs with large and small pupil, respectively.
G. Pattern similarity during water cue trials in ‘Thirsty’, ‘Quenched’, and ‘Quenched+SFO^GLUT activation’ states. Gray shaded areas: cue period. Mean±s.e.m across 4 mice.
H. Similar to G, but for food cues across ‘Hungry’, ‘Sated’, and ‘Sated+AgRP activation’ states. Gray shaded areas: cue period. H: hungry; S: sated. Mean±s.e.m across 4 mice.

Flowchart of Thirsty Choose Water study

Flow of participants through the stages of the Thirsty for a Smile study.