Fig. 5

Loss of C3aR in the Smarca5 cKO cerebellum results in increased Bergmann glia disorganization. At P1, BLBP labeling in mutant cerebellums showed the cell bodies of the Bergmann glia to be mislocalized (a), being frequently positioned immediately adjacent to the EGL (arrows; the EGL is indicated with a dashed line). BLBP labeling was further altered in the dKO samples in that it was weak throughout the cerebellum, except within the EGL, where it labeled strongly. Iba1⁺ cells were also labeled to look for any correspondence in Bergmann glia disorganization and phagocyte localization, which was not observed. In P10 samples, the orderly array of Bergmann glia cell bodies within the Purkinje cell layer observed in control samples was mostly absent in both Smarca5 cKO and dKO samples. Both of these mutants also displayed increased GFAP labeling and aberrant arborization, which was worsened in the dKO sections. Within fissures of the cerebellum (b), the nearly absent EGL and strongly GFAP⁺ and disorganized processes of the Bergmann glia in the P10 dKO contrasted with a more normal-appearing Smarca5 cKO single mutant. Calbindin labeling identified the Purkinje cells. c Transcript expression for GFAP was increased in both of the Smarca5 cKO and dKO cerebellums, being higher in the dKO mutants (n = 4 mice/genotype; **p < 0.005, ***p < 0.0001). Transcripts for the inflammatory cytokines IL6 and TNF were not increased (IL6, n = 4–5 mice/genotype; TNF, n = 5 mice/genotype). d Total GFAP protein was increased in both the Smarca5 cKO and dKO cerebellums relative to cerebellums from controls and C3aR KO mice. e Quantification of the GFAP/BLBP ratio from the immunoblots shown in D (**p < 0.01; ***p < 0.001). Images in a and b were reconstructions of optical sections. Scale bar in a = 40 μm, and applies to all panels; scale bar in b = 40 μm, and applies to both panels