Figure 2
Assessment of microglial cells in lumbar spinal cord of SOD1G93A mice.
(a–c) FACS analysis from lumbar spinal cord of WT, SOD1G93A untreated and treated with GW2580 showing microglial cell counts (CD45low, CD11b+) at 12 and 16 weeks. (n = 4 animals per group and time point). (b,c) Representative density FACS plots showing the reduction of microglia in 16 weeks old SOD1G93A mice after GW2580 treatment. (d–f) Representative micrographs of L4-L5 ventral horns stained for Iba1 from (d) WT, (e) SOD1 control or (f) GW2580-treated SOD1 mice. (g) Graph showing immunohistochemical quantification of Iba1+ cells in the ventral horns of spinal cord tissue sections of 16 weeks of age SOD1G93A mice untreated or treated with GW2580 (n = 4 per group). Note, that immunohistochemical analysis highly correlates with counts obtained by FACS analysis. (h) Quantification of microglial proliferation with EdU assay by FACS analysis. Note that selective blockade of CSF1R reduces proliferation of microglial cells in SOD1G93A mice at 16 weeks of age (n = 4 animals per group and time point). (i,j) Representative images of microglial proliferation by double immunofluorescence for EdU (red), Iba1 (green) and DAPI (blue) in the ventral horn of L4-L5 spinal cord tissue sections of (i) SOD1G93A control and (j) treated with GW2580 at 16 weeks of age. Note that CSF1R blockade significantly reduces microglia proliferation but only at 16 weeks of age. Scale bars: (d–f) 200 μm; (i,j) 50 μm. *p < 0.05 compared to SOD1G93A untreated, #p < 0.05 compared to WT. Error bars indicate SEM.
