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. 2022 Mar 17;219(4):e20202304. doi: 10.1084/jem.20202304

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© 2022 Kurematsu et al.

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Figure 3. — PS exposure on less-active spines, and its suppression by olfactory inputs in the adult OB. (A and B) Representative images c-Fos staining (red) of the GCL in the naris-occluded (A) and opened (B) side of OB in adult AiCE-Tg mice (n = 3 mice) with unilateral naris occlusion. Nuclei were stained with Hoechst 33342 (blue). Arrows indicate c-Fos⁺ signals in the GCL. (C) Density of the c-Fos⁺ cells in the GCL (n = 3 mice, pooled from two independent experiments, t₍₂₎ = −7.7, P = 0.016, paired t test). (D and E) Representative images of the EPL in the naris-occluded (D) and opened (E) side of OB in the adult AiCE-Tg mice (n = 3 mice) stained for PS (white), PSD95 to define spines (red), and EGFP (green, EGFP-CapZ). Active and less-active spines, defined here by the level of EGFP signal (see Materials and Methods), in E are enlarged and shown by orthogonal view in (E¹ and E²) and (E³ and E⁴), respectively. Arrowheads and arrows (E and E¹–E⁴) indicate active spines and representative PS⁺ spines, respectively. (F) Representative single z-plane and orthogonal images of the active spine (arrowhead) in the EPL of the naris-opened side of the OB of adult AiCE-Tg mice (n = 3 mice) stained for the postsynaptic marker PSD95 (red), EGFP (green, EGFP-CapZ), and a presynaptic marker Bassoon (white). (G) Proportion of PS⁺ population in active and less-active PSD95⁺ spines in the EPL of the adult OB (pooled from two independent experiments; 0 PS⁺/629 active spines, 282 PS⁺/44,371 less-active spines from two mice; P = 0.037, Fisher’s exact test). (H–J) Representative images of the EPL in the OB of naris-opened (H, n = 4 mice), naris-occluded (I, n = 4 mice), and odor-enriched (J, n = 6 mice) adult WT mice stained for PS (white) and PSD95 (red). PSD95⁺PS⁺ spines (arrows) are enlarged and shown by orthogonal view in the boxes (H¹, H², I¹, I², I³, and J¹). Different z-level images of PSD95⁺PS⁺ spine in (H¹ and I¹) are also shown by a single z-plane and orthogonal view (z + 0.5 µm). (K) Density of the PSD95⁺PS⁺ spines (n = 4 [opened], 4 [occluded], 6 [odor-enriched] mice pooled from two independent experiments; F_(2,11) = 33.3, P = 0.000022, one-way ANOVA; opened versus occluded, P = 0.0041, opened versus enriched, P = 0.011, occluded versus enriched, P = 0.000015, Tukey–Kramer test). GCL, granule cell layer; EPL, external plexiform layer. Scale bars: A, B, 20 µm; D–F and H–J, 2 µm. *, P < 0.05; ***, P < 0.005. Data shown are mean ± SEM.