Fig. 4
Quantification of cellular density into scala tympani of the base of cochlea following cochlear implantation. Cochlear implantation was performed in CX3CR1+/GFP Thy1+/YFP mice, fed on chow with 1200 ppm of PLX-5622 (PLX) or control chow (No PLX) for 7 days. Treatment with respective diets and electrical stimulation was continued until the desired endpoints (10, 28, or 56 days). Nuclei were labeled with Hoechst 3342 in 30-µm-thick mid-modiolar sections. Image analysis was performed in IMARIS image analysis software. Hoechst + cells were counted on maximum intensity z-projections of 3D confocal image stacks in scala tympani of the base of the cochlea. The volume of scala tympani and nuclear counts were made using an automated counting system in IMARIS image analysis software aided by custom-made macros. Nuclear density (Hoechst + cells) was calculated per 105 µm3. An average of 3 sections per animal was taken with ‘n’ being the number of animals. Number of cochlea analyzed in this study are as follows: at day 10 post-CI, Implanted ‘no PLX’ (n = 4), implanted PLX, (n = 3), contralateral ‘no PLX’ (n = 3), contralateral PLX (n = 5); at day 28 post-CI, Implanted ‘no PLX’ (n = 6), implanted PLX, (n = 5), contralateral ‘no PLX’ (n = 3), contralateral PLX(n = 4); at day 56 post-CI, Implanted ‘no PLX’ (n = 3), implanted PLX, (n = 6), contralateral ‘no PLX’ (n = 3), contralateral PLX (n = 4). Error bars indicate SEM