Fig. 4

Pharmacological mGlu3 receptor blockade and microglial reactivity in response to inflammatory stimulation in vitro. a, b Microglial cells sorted from P4 pups were stained with Iba1 (green) and DAPI (blue) under basal conditions ± LY 2389575 (5 μM). Representative photomicrographs at × 40 magnification are shown in a (scale bar = 50 μm). The cell area, cell perimeter, and cell circularity were assessed in b (mean cell number, 162 ± 10). Data (mean ± SEM). Unpaired t test; **p < 0.01. c, d mRNA expression of pro-inflammatory (c) and anti-inflammatory/immune-regulatory (d) markers in microglial cells sorted from P4 pups cultured under basal or pro-inflammatory (IL-1β + INFγ) conditions ± LY 2389575 (5 μM). The data (mean ± SEM) are relative to the gene expression under basal CTRL conditions. One-way ANOVA followed by the Newman-Keuls multiple comparison test; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001 effect of LY 2389575; ^$p < 0.05, ^$$p < 0.01, ^$$$p < 0.001, ^$$$$p < 0.0001 effect of IL-1β + INFγ. e, f Microglial cells sorted from P7 pups were stained with Iba1 (green) and DAPI (blue) under basal conditions ± LY 2389575 (5 μM). Representative photomicrographs at × 40 magnification are shown in e (scale bar = 50 μm). The cell area, cell perimeter, and cell circularity were assessed in f (mean cell number, 162 ± 10). Data (mean ± SEM). Unpaired t test *p < 0.01, **p < 0.01. g, h mRNA expression of pro-inflammatory (g) and anti-inflammatory/immune-regulatory (h) markers in microglial cells sorted from P7 pups and cultured under basal and pro-inflammatory (IL-1β + INFγ) conditions ± LY 2389575 (5 μM). The data (mean ± SEM) are relative to the gene expression under basal CTRL conditions. One-way ANOVA followed by the Newman-Keuls multiple comparison test *p < 0.05, **p < 0.01, ***p < 0.001 effect of LY 2389575; ^$$p < 0.01, ^$$$p < 0.001, ^$$$$p < 0.0001 effect of IL-1β + INFγ