Fig. 9

Transplantation of iNSCs alters oligodendrocyte populations during recovery. a: Tissues from the cohorts with Miss-step wheel access (Figs. 7 and 8) were further analyzed based on Plp1 transcripts as either low, perinuclear signal (Plp1^low) or high and extending into processes (Plp1^high). iNSC transplantation increased Plp1^low cells during remyelination with wheel running access. b-c: Nuclear Ki67 identified rare proliferating cells. iNSC transplantation increased the proportion of Ki67 cells immunolabeled for the NG2, a marker of OPCs, in the corpus callosum ipsilateral (ipsi) to iNSC/vehicle injections (b). Example of a pair of proliferating OPCs (c, arrows) co-labeled for NG2 and nuclear Ki67 (overlap appears yellow) and quiescent OPCs (c, arrowheads). d-e: Analysis of additional mice without wheel access shows reduced Plp1^low cells (d) and increased Plp1^high cells (e) during remyelination after chronic CPZ. This difference indicates that Plp1^low and Plp1^high cells represent distinct lineage stages and/or responses after chronic CPZ. f-h: Cohorts using acute CPZ demyelination to induce extensive remyelination with and without wheel exposure. Without wheels, CPZ decreased Plp1^low cells (f) while increasing Plp1^high cells (g) as well as newly forming oligodendrocytes identified high expression of Enpp6 (Enpp6^high) (h). Miss-step wheel access during remyelination simultaneously reduced Enpp6^high cells and increased Plp1^low cells (f, h). i: Model integrating expression of Plp1 and Enpp6 transcripts during remyelination after acute and chronic CPZ with iNSC and wheel effects. For oligodendrocyte graphs, bar color reflects condition: no cuprizone (white), cuprizone without wheels (black), cuprizone with wheels (blue), and mice with iNSC (green). Data are mean values ± sem. For chronic CPZ mice from the wheel running cohort, two-way ANOVA was used with Sidak’s multiple comparisons test of CPZ vehicle (n = 10) versus CPZ iNSC (n = 9). Ki67 and NG2 data analyzed using a contingency table with Fisher’s exact test for CPZ vehicle (n = 3) versus CPZ iNSC (n = 3). Chronic CPZ mice without wheel exposure had naïve (n = 5) and CPZ (n = 5) conditions compared by t-tests. The acute CPZ study compared conditions of naïve (n = 6; n = 5 for Enpp6) and CPZ (n = 6) mice with wheel exposure and CPZ (n = 6) mice without wheels using one way ANOVA with Sidak’s multiple comparisons test