Extended Data Fig. 6: Fear conditioning with contextual novelty induces astrocyte Fos ensembles.
From: The astrocytic ensemble acts as a multiday trace to stabilize memory
a, Left: Whole-brain mapping pipeline. Alongside NoFC and FC groups (Fig. 1a), an additional cohort received 4-OHT immediately after CtxA exposure with shocks but no habituation (FC+Novelty). Brains were fixed and processed for STPT, 3D reconstruction, regional segmentation, and single-cell counting as in Fig. 1a. b, 3D rendering of the whole brain and three brain regions - the hippocampus (HPC; CA1, CA2, CA3 and dentate gyrus), the amygdala (lateral, basolateral and basomedial amygdalar subregions), primary visual cortex (VISp) - showing that Fos-mNG astrocytes (white dots). D dorsal, V ventral, A anterior, P posterior, M medial, L lateral. Scale bars, 500 µm. c-d, Heatmap of BAE count fold changes (vs NoFC in c, vs FC in d, n = 4 mice/group). *FDR < 0.05. *FDR < 0.05; full stats in Supplementary Table 2. Levene’s test followed by one-way ANOVA or Kruskal-Wallis; post-hoc Tukey or Dunn with Bonferroni. FDR controlled by Benjamini-Hochberg. Full data in Supplementary Table 2. Primary visual area (VISp), primary somatosensory area barrel field (SSp-bfd), orbital cortex (ORB), prelimbic cortex (PL), infralimbic cortex (ILA), entorhinal cortex (ENT), anterior cingulate cortex (ACA), olfactory cortex (OLF), retrosplenial cortex (RSP), lateral amygdala (LA), basal amygdala (BA), central amygdala (CEA), mediodorsal thalamus (MD), dorsal geniculate thalamus (GENd), anterodorsal thalamus (AD), anteroventral thalamus (AV), ventroposterior medial nucleus of thalamus (VPM), caudoputamen (CP), nucleus accumbens (ACB), dentate gyrus (DG), hippocampal CA1 (CA1), hippocampal CA3 (CA3), para-subiculum (PAR), and pre-subiculum (PRE). e, Hippocampus/CA1 BAE counts higher in FC+Novelty. FC-, Novelty-: n = 6 mice, FC+, Novelty-: n = 4 mice, FC+, Novelty+: n = 4 mice. Data are mean ± SEM. *p < 0.05. Sample sizes and replicates in Supplementary Table 1.
