(A) Top: Photometric traces of calcium responses in the PVT
to 10 repetitions of randomized odor (Left) and water (5μl, Right)
stimuli. Dashed line indicates the time of stimulus delivery. Scale bar, 10%
ΔF/F, 3 s. Bottom: Left Y axis, quantification of odor (black dot) and
free water (red dot) evoked ΔF/F over 10 repetitions. Right Y axis,
quantification of free water (orange circle) evoked licks over 10 repetitions.
Novel odor: n = 6 mice; Free water n = 12 mice; Licks: n = 12 mice.
(B) Trial structure of the Pavlovian conditioning paradigm.
ITI, inter-trial interval. CS: conditioned stimulus; US: unconditioned stimulus.
(C) Mean lick rate of well-trained animals (n = 7) shows
anticipatory licking in appetitive (blue) but not neutral (black) and aversive
(red) trials. Gray bar: 1s of CS delivery; Vertical dash line: US delivery.
(D) Mean photometric responses of the PVT to CS and US in both
appetitive (blue) and aversive (red) but not neutral (black) trials. Shade, SEM
across mice, n = 7 mice. (E) Z score heat maps (left) and pie chart
(right) for all task-responding neurons identified by in vivo
single-unit recording during Pavlovian tasks of well-trained animals. Neurons
are separated in five subgroups based on their tuning properties, and are
rank-ordered by their response onset times during reward cue stimulation. Each
row in the heat maps represents responses from the same neuron to different
stimuli. n = 85 neurons from 12 mice. (F, G) Mean photometric
responses (F, n = 7 mice) and quantification (G)
showing that CS and US response in the PVT are graded to different intensity of
reward (left, 5 vs 15 μl water) and punishment (right, air puff vs. tail
shock). AUC, area under curve (see methods). Scale bar, 2% (F,
left), 4% (F, right) ΔF/F, 1s. Big, small reward and
nothing: n = 7,7,7; Big, small punishment and nothing: n = 6,6,5.
*P < 0.05, **P < 0.01
(One-way ANOVA, Post-hoc Tukey’s test). Shade, SEM across mice in
C, D, F. Data are means ± SEM.