Figure 2. Disruption of NR1 in oligodendroglial lineage cells does not alter EAE disease course, nor interrupt neonatal OPC maturation and differentiation. A, Five days of TM treatment reduces severity of EAE if MOG-peptide immunization is only 10 d post-TM (insert). B, Mean clinical scores of PRG+TM mice did not differ from those in PRG+V and PR+TM mice. C, Cumulative disease index (summed clinical score) on day 51 post-MOG immunization is similar in PRG+TM, PRG+V and PR+TM mice. D, E, Representative confocal images showing SMI312+ axons in dorsal column and MBP+ myelin in PRG+TM and PRG+V mice on day 51 post-MOG immunization. F, RT-qPCR showing the absence of NR3A mRNA in NR3A−/− mice, and that NR1 mRNA levels were similar in these three groups. Note that two primer sets targeting NR3A confirmed the lack of mRNA transcripts in NR3A KO mice. G, H, NR3A KO mice show similar EAE course (G), and similar cumulative disease index (H) to their littermates. I, Percentage of EYFP+ cells that are CC1+ mature oligodendrocytes and NG2+ OPCs in forebrain fimbria (Fim) and hippocampus (Hip) on P15 of PR and PRG mice that received TM on P7. J, K, Orthogonal confocal images showing colocalization (arrowheads) of EYFP and MBP (J) and Opalin (K), a protein expressed in myelinating oligodendrocytes, in fimbria on P15 PRG mice that received TM on P7. Note MBP is mainly restricted to EYFP+ processes, whereas Opalin is in both cell body and processes (arrowheads). L, PCR data confirming the Grin1 deletion in CNP-Cre, Grin1flox/flox NR1 KO but not in Grin1flox/flox non-KO mice (same strategy as in Fig. 1A). M, N, Sox10+/NG2+ OPCs and Sox10+/CC1+ mature oligodendrocytes in the corpus callosum on P22 mice. Arrows and arrowheads point to Sox10+/CC1+/NG2− mature oligodendrocytes and Sox10+/CC1−/NG2+ OPCs, respectively. O, Western blotting showing MBP expression in P22 forebrain between NR1 KO and non-KO groups. Scale bars: D, E, J, K, 10 μm; M, 25 μm.