Fig. 1 |. Consumption of a novel flavor supports one-shot CTA learning and activates different brain regions than the same flavor when familiar.

a. Schematic of the CTA paradigm. b, Flavor preference across three consecutive daily retrieval tests for mice that consumed either a novel (top) or familiar (bottom) flavor and then were injected with either LiCl (red) or saline (black) on pairing day (n = 8 mice per group). The specific flavor (sweetened grape kool-aid) and amount consumed (1.2 ml) was the same for all groups. The familiar group was pre-exposed to the flavor on four consecutive days before conditioning, whereas the novel group was completely naïve. c, Schematic and example Fos expression data (100-μm maximum intensity projection) for the brainwide light sheet imaging pipeline. d, Schematic of the Consumption Fos timepoint (n = 12 mice per group). The line above “10 min” is a scale bar and the gray bar represents the 60-min wait before perfusion. e, Novel flavors preferentially activate sensory and amygdala regions. Left: Comparison of individual familiar (blue) and novel (red) flavor condition mice for every significantly novel flavor-activated brain region. Right: Visualization of the spatially resolved difference in Fos⁺ cell density across flavor conditions with Allen CCF boundaries overlaid. f, Familiar flavors preferentially activate limbic regions. Panels are analogous to e. P-values in b are from GLMM marginal effect z-tests corrected for multiple comparisons across retrieval days within each flavor condition. P-values in e,f are from GLMM marginal effect z-tests corrected for multiple comparisons across timepoints within each brain region. Error bars represent mean ± s.e.m. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. See Extended Data Table 1 for list of brain region abbreviations.