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. 2020 Dec 9;108(5):887–904.e12. doi: 10.1016/j.neuron.2020.09.010

Figure 5.

Figure 5

Acute APAP Treatment Influences Microglia Activity, and TrpV1 Channels Acutely Regulate Microglia Activation in Dp(16) Animals

(A) Experimental protocol.

(B) Voltage-activated current profile in microglia of WTCX3CR1-GFP and Dp(16)CX3CR1-GFP hippocampal slices (left inset) treated acutely with vehicle or APAP. Voltage currents were analyzed using ramps ranging from −100 to 80 mV (right inset). Data are expressed as average current (picoAmpere,pA) normalized for the capacitance (picoFarad,pF) for each mV step of all analyzed cells ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; two-way RM ANOVA; FInteraction (9, 315) = 10.55; p < 0.001; Holm-Sidak post hoc test. In parenthesis: analyzed cells, animals.

(C and D) Quantification of the mobility index and process speed of microglial cells. Bars represent the average mobility index and process speed in all analyzed fields ± SEM, and circles represent data points for each field (3 animals per genotype). ∗∗∗p < 0.001; (C) two-way ANOVA, FInteraction (1, 43) = 15.07, p < 0.001; Holm-Sidak post hoc test; (D) two-way ANOVA, FInteraction (1, 225) = 31.25, p < 0.001; Holm-Sidak post hoc test. The vehicle-treated cell data (dotted circles) are from Figures 1K and 1L for comparison.

(E–G) Voltage-activated current profile in microglia of WTCX3CR1-GFP and Dp(16)CX3CR1-GFP hippocampal slices treated with vehicle or the indicated drug (OLDA, 2 μM; AM-251, 1 μM; AM-630, 1 μM; capsazepine, 10 μM). Voltage currents were analyzed using ramps ranging from −100 to 80 mV, as in the inset. Data are expressed as average current (picoAmpere,pA) normalized for the capacitance (picoFarad,pF) for each mV step of all analyzed cells ± SEM. ∗ represents the statistical difference between Dp(16) and WT vehicle, ∗p < 0.05, ∗∗∗p < 0.001; § represents the statistical difference between Dp(16) vehicle and Dp(16) treated, §p < 0.05, §§p < 0.01, §§§p < 0.001; (E) two-way RM ANOVA, FInteraction (27, 1,341) = 14.35, p < 0.001; Holm-Sidak post hoc test; (F) two-way RM ANOVA, FInteraction (27, 1,449) = 20.34, p < 0.001; Holm-Sidak post hoc test; (G) two-way RM ANOVA, FInteraction (27, 1,580) = 18,89, p < 0.001; Holm-Sidak post hoc test. In parenthesis: analyzed cells, animals.

(H) Cartoon depicting the putative mechanism of action of APAP. FAAH, fatty acid amide hydrolase.