Extended Data Fig. 10: NLOT’s participation in CFC, and diversity in molecular NLOT types and projections.
From: Neuronal types in the mouse amygdala and their transcriptional response to fear conditioning
a, Pearson pairwise correlation (green, low; yellow, high) of 18 IEGs (rows and columns) for VGLUT1-8, resolved by post-CFC sampling time point and batch-specific home cage (HC) control. Top row, batch B (HC-2h-recall); bottom row, batch A (HC-8h-24h); genes are ordered by hierarchical clustering, indicated on the right. b, Expression of retro-AAV-hSyn1-GCaMP6f (cyan) in the injection site (left; MOB, with accessory olfactory nucleus (AON) strongly labeled), and recording site (right; NLOT) of one mouse. NLOT zoom-ins, 3 a.-p. positions reveal GCaMP6f expression in layer 1 and 2 NLOT, and tissue damage (outlined) indicates proximal optic-fiber tip placement. c, Examples of individual calcium-traces in three consecutive tone-shock pairings; with tone and shock duration indicated. d,e, NLOT projection tracing by injection of retro-AAV-hSyn1 to MOB (GFP; cyan) and BLA (mCherry; magenta). MOB-projecting NLOT neurons (cyan) primarily populate ipsi- and contralateral NLOT layer 2, BLA-projecting NLOT neurons (magenta) dominate layer 3; both populations partially express Sim1 (e). f, Spatial correlation to three Visium ST sections, for NLOT populations VGLUT1 6-8. g, Gene expression visualized as single-cell violin plots. h, Two additional populations sparsely label NLOT layer 3, but also BLA-v; indicated by inferred spatial distributions (AMBA volumetric correlation), and individual marker genes in Allen Mouse Brain Atlas (Bok, https://mouse.brain-map.org/experiment/show/71064032; Pamr1, https://mouse.brain-map.org/experiment/show/70744322; Lamp5, https://mouse.brain-map.org/experiment/show/70927827; Thrsp, https://mouse.brain-map.org/experiment/show/71587845). i, NLOT projection tracing (as d,e) in posterior sections reveal MOB-projecting (but not BLA-projecting) NLOT neurons (cyan) also populate NLOT layer 1, and putative bed nucleus of the accessory olfactory tract (BA). j, Individual genes expressed in NLOT layer 1 and BA (bed nucleus of the accessory olfactory tract), from Allen Mouse Brain Atlas (Slc17a7, https://mouse.brain-map.org/experiment/show/70436317; Slc17a6, https://mouse.brain-map.org/experiment/show/73818754; Gad2, https://mouse.brain-map.org/experiment/show/79591669; Lhfp, https://mouse.brain-map.org/experiment/show/73769323; Trh, https://mouse.brain-map.org/experiment/show/71016631).