Fig. 8

Post-behavior analysis of CC remyelination and endogenous glial cell response. a-b: Coronal CC sections immunolabeled for MOG to detect myelin in the wheel cohort mice after chronic CPZ with vehicle (a) or iNSC (b) injections. c-d: Innate immune cell response examined with immunolabeling for GFAP (c) or (d). Panel C shows astrogliosis and transplanted iNSCs (green, GFP) within the CC and adjacent areas in a section containing the region of the injection needle track. Panel D shows higher magnification of an iNSC (green, GFP; white arrow) in the CC with thin processes. IBA1 immunolabeled cells contain lipofuscin granules (yellow autofluorescence; yellow arrows). iNSCs did not immunolabel for IBA1. e-f: Coronal CC sections processed for in situ hybridization to detect oligodendrocytes expressing proteolipid protein (Plp1). Plp1 appears upregulated after iNSCs transplantation (f) compared to vehicle (e). Plp1 transcripts are localized in the cell body in most cells while a subset of cells show more intense labeling in the cell body and extending into processes. g-k: Quantification of tissue analysis in this post-behavior cohort. CC atrophy and demyelination are not attenuated by iNSCs (g, h). Transplanted iNSCs did not reduce astrogliosis (i) or the microglial response (j). In contrast, the iNSCs increased the oligodendrocyte population (k). Bar color reflects condition as chronic cuprizone followed by vehicle injection and wheels (blue) or iNSC transplant and wheels (green). Data are mean values ± sem. Statistical analysis used t-tests to compare vehicle and iNSC conditions. For immunolabeling (MOG, GFAP, IBA1), vehicle n = 12 and iNSC n = 11. For Plp1 in situ hybridization, vehicle n = 10, iNSC n = 9. Scale bars: A, B, shown in A = 200 μm; C = 100 μm; D = 20 μm; E, F, shown in E = 50 μm