Fig. 2

DRD3 specific immunoreactivity is associated to astrocytes in the mouse brain upon systemic inflammation. Wild-type mice received an i.p. injection of PBS or LPS (5 mg/kg) and 3 h later were sacrificed and immunofluorescence analysis was performed in brain slices. To determine the specific DRD3 immunostaining, a polyclonal anti-DRD3 antibody was pre-incubated or not with the peptide DRD3_15-29 (CGAENSTGVNRARPH) used as immunogen to develop the antibody. Afterward, brain sections were incubated with untreated anti-DRD3 or with anti-DRD3 pre-incubated with peptide DRD3_15-29, followed by incubation with the AlexaFluor488-coupled secondary antibody (green). Samples were subsequently immunostained with anti-GFAP-PE (red) antibody and nuclei were stained with DAPI (blue). a Representative images of the subventricular zone are shown (top panels; bar, 5 μm). Pseudocolored images showing areas in which DRD3-associated intensity correlates positively with GFAP-associated intensity are shown in bottom panels (bar, 5 μm). b Intensity correlation analysis of confocal microscopy images obtained from DRD3 and GFAP immunostaining. Colocalization intensity was determined as the intensity correlation quotients (ICQ). Data from four mice per group is shown. Values are the mean ± SEM. *p < 0.05; **p < 0.01; by two-way ANOVA followed by Sidak’s post-hoc test