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. Author manuscript; available in PMC: 2021 Jul 16.
Published in final edited form as: Cell Rep. 2021 Jul 6;36(1):109313. doi: 10.1016/j.celrep.2021.109313

Figure 2. Microglia interact with PNN-coated neurons upon repeated ketamine stimulation.

Figure 2.

(A) Immunostaining for Iba1 (green) and parvalbumin (Pvalb, cyan) after 3× saline or KXA in primary somatosensory cortex S1. Zoom-in, single-plane orthogonal projection. Arrow, microglia in close distance of Pvalb+ neurons. Scale bar: 10 μm. (B) Minimal Pvalb+ neuron-microglia median distance in μm after treatment. Each condition 3 animals as indicated in legend, 5 microglia per animal. Linear regression model with animal ID as random effect: P♀ < .0001, P♂ < .0001. (C-D) Ex-vivo live imaging of brain slices of Cx3Cr1CreERT2 het/Ai9 het mice dissected 2-hours after 2× KXA. Snapshots of microglia (green) and WFA (magenta) with orthogonal side projection. Scale bar: 10 μm. (D) Time course of mean WFA volume within microglia percentage ± SEM after treatment. Linear regression model: P < .0001 with selected post-hoc comparisons. (E-F) Mean percentage CD68 volume within microglia ± SEM after (E) saline or KXA or (F) 3× KXA recovery. See also Figures S5A and S5B. Linear regression model: (C) P < .0001, (D) P < .0001 with selected post-hoc comparisons. (G) Mean percentage of PNN volume within microglial CD68 ± SEM upon treatment. Linear regression model: P < .0001 with selected post-hoc comparisons. Each dot represents one animal (3–5 animals per condition with exception of the control bar in (E) and (G), which shows the mean value of all controls. See also Figures S5A and S6B, respectively). (H) Immunostaining for PNN (WFA, magenta), endosomal-lysosomal marker CD68 (blue), and microglia (Iba1, green) after 2× KXA. Below: single plane orthogonal projections. Scale bar: 5 μm. ***P < .001, **P < .01, nsP > .05.