Figure 2

Diminished microglia cell numbers and extent of CD68 ⁺ content in CXCR3-/- cuprizone brains. A. Sagittal brain sections of wild type (WT) and CXCR3-/- were analyzed for the localization of activated microglia at regions of the corpus callosum (Cc), thalamus (Th) and cerebellum (Cb). B. Analysis of serial sagittal brain sections stained for tomato lectin documents a significant reduction of activated microglia in cuprizone-fed CXCR3-/- mice compared to WT animals (n = 10 to 14 at equal sections, time point and genotype, *P <0.05, **P <0.005, ***P <0.001). D. These observations were made within regions of the thalamus (Th) and cerebellum (Cb), but were not found in the white matter of the frontal corpus callosum (Cc) of CXCR3-/- brains. Besides the reduced lectin⁺ area fraction in the cerebellum of CXCR3-/- mice, we also found a differential spatial distribution of microglia surrounding the cerebellar nuclei together with a consistent cluster at the vestibular nuclei of the medulla after 5 weeks (D, WT Cb versus CXCR3-/- Cb). {AU Query: Please define ‘D’ from the previous sentence so readers know what this abbreviation represents.} Because most overt differences of microglia accumulation between CXCR3-/- and WT were visible after 5 weeks of cuprizone diet, we further investigated microglia using double fluorescence immunolabeling for CD68 (red) and MBP (green) (Myelin basic protein). C. At the designated insets (D) we analyzed the CD68⁺ content in both genotypes and found significantly reduced particle perimeters in all studied central nervous system (CNS) regions in CXCR3-/- compared to WT brains (n ≥1,900 CD68⁺ particles per genotype, **P <0.01, ***P <0.001). Data represent mean ± SEM.