3.0.CO;2-R" data-track-action="article reference" href="https://doi.org/10.1002%2F%28SICI%291096-9861%2820000410%29419%3A3%3C364%3A%3AAID-CNE8%3E3.0.CO%3B2-R" aria-label="Article reference 7" data-doi="10.1002/(SICI)1096-9861(20000410)419:33.0.CO;2-R"Article CAS Google Scholar /p> 0.9999 and 0.7698, Mann Whitney and 2-tailed unpaired Student's t-test, respectively. At 3-4 months, n = 5 FIRE+/+ and 4 FIREΔ/Δ mice, at 6 months, n = 5 FIRE+/+ and 3 FIREΔ/Δ mice. d) Flow cytometry gating strategy for assessment of non-microglial myeloid cells (including BAMs; CD11b+ CD45hi) for panels (e) and (f). e) Intensity of expression of CSF1R in FIRE+/+ and FIREΔ/Δ mice in CD11b+CD45lo microglia (MG) versus CD11b+CD45hi myeloid cells. f) Mean Fluorescence Intensity (MFI) of CSF1R ± s.e.m. in non-microglial myeloid cells (including BAMs) in FIRE+/+ and FIREΔ/Δ mice. ****P < 0.0001, 2-tailed unpaired Student's t-test. n = 4 FIRE+/+ and 5 FIREΔ/Δ mice g) Images of astrocytes (SOX9; green; GFAP+; magenta) at 1 month of age in the corpus callosum. Scale bar, 75 μm. Inset shows magnified view of double positive cells. Scale bar, 25 μm. h) Mean SOX9+ GFAP+ cells/mm2 ± s.e.m. at 1 month of age in the corpus callosum. n = 5 mice per group. Non-significant, P = 0.4799, 2-tailed unpaired Student's t-test./p> 0.9999, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. c) Mean primary errors ± s.e.m. during training phase. n = 13 FIRE+/+ mice and 10 FIREΔ/Δ mice. Non-significant, Day 1: P = 0.9649; Day 2: P = 0.5968; Day 3: P = 0.8884; Day 4: P = 0.6028; Day 5: P = 0.9215; Day 6: P = 0.9437, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. d) Mean percentage of time spent in the target quadrant± s.e.m., n = 13 FIRE+/+ and 10 FIREΔ/Δ. Dotted line indicates 25% chance. Non-significant between genotypes, 1hr: P = 0.7337; 3d: P > 0.9999, 2-way ANOVA with Sidak's multiple comparisons test. e) Mean number of nose pokes into holes during 1hr probe test ± s.e.m., n = 13 FIRE+/+mice and 9 FIREΔ/Δ mice. Target **P = 0.0019, 2-way ANOVA with Sidak's multiple comparisons test. f) Mean number of nose pokes into holes during 3d probe test ± s.e.m., n = 13 FIRE+/+mice and 9 FIREΔ/Δ mice. Non-significant, P = 0.9329, 2-way ANOVA with Sidak's multiple comparisons test. g) Reversal phase: Target hole 2 (blue) is 180° from the original target; mice require cognitive flexibility to adapt to the new target. h) Mean primary latency (sec) ± s.e.m. over 3 training days. n = 13 FIRE+/+ and 9 FIREΔ/Δ. Non-significant across all training days between genotypes, Day 1: P = 0.9999; Day 2: P = 0.5186; Day 3: P = 0.9622, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. i) Mean primary errors ± s.e.m. during reversal days. n = 13 FIRE+/+ and 9 FIREΔ/Δ. Day 1: *P = 0.0488, Day 2: *P = 0.0142, Day 3: P = 0.6727, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. j) Mean percentage of time spent in the target quadrant during reversal probe test ± s.e.m. n = 13 FIRE+/+ mice and 9 FIREΔ/Δ mice. Dotted line indicates 25% chance. Non-significant, P = 0.6933, 2-tailed unpaired Student's t-test. k) Mean number of nose pokes into holes during reversal probe test ± s.e.m. n = 13 FIRE+/+ mice and 9 FIREΔ/Δ mice. Non-significant P = 0.9657, 2-way ANOVA with Sidak's multiple comparisons test. l) Schematic of Open Field test. m) Total distance travelled (m) during 10-min Open Field test ± s.e.m. n = 37 mice: n = 15 FIRE+/+ mice (7 young, 8 middle-aged); n = 22 FIREΔ/Δ mice (12 young, 10 middle-aged). Young mice = 4–8 weeks of age, Middle aged mice = 11–13 months of age. Non-significant, Young: P = 0.9997; Middle-aged: P > 0.999, 2-way ANOVA. n) Percentage (%) of time spent in the centre of the arena during Open Field test ± s.e.m. n = 37 mice: n = 15 FIRE+/+ mice (7 young, 8 middle-aged); n = 22 FIREΔ/Δ mice (12 young, 10 middle-aged). Young mice = 4–8 weeks of age, Middle aged mice = 11–13 months of age. Non-significant, Young: P = 0.7899; Middle-aged: P > 0.9995, 2-way ANOVA. o) Mean distance travelled (m) during training days ± s.e.m. n = 13 FIRE+/+ mice and 9 FIREΔ/Δ mice. Non-significant, P = 0.9607, >0.9999, >0.9999, 0.9955, 0.6583, 0.6034, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. p) Mean distance travelled (m) during reversal days ± s.e.m. n = 13 FIRE+/+ mice and 9 FIREΔ/Δ mice. Non-significant, P = 0.7856, 0.9784, 0.8626, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. q) Mean speed (m/s) during training days ± s.e.m. n = 13 FIRE+/+ mice and 9 FIREΔ/Δ mice. Non-significant, P = 0.9998, >0.9999, 0.6667, 0.9995, 0.9831, 0.9995, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test. r) Mean speed (m/s) during reversal days ± s.e.m. n = 13 FIRE+/+ mice and 9 FIREΔ/Δ mice. Non-significant, P = > 0.9999, 0.9940, 0.1561, Repeated measures 2-way ANOVA with Sidak's multiple comparisons test./p> 0.9999; CC1-: P > 0.9999, 1-way ANOVA with Tukey's multiple comparisons test. d) Representative images of EdU+ (magenta) CC1+ (green) OLIG2+ (white) triple positive cells (arrows). Scale bar, 25 µm. e) Images of corpus callosum 6 weeks post cognitive testing. Scale bar, 1 µm. f) Mean number of myelinated axons/mm2 ± s.e.m. in untrained versus trained mice. n = 3 mice/group in untrained category, 4 trained FIRE+/+ mice and 6 trained FIREΔ/Δ mice. FIRE+/+ mice *P = 0.0364, FIREΔ/Δ mice non-significant, P = 0.8537, 2-tailed unpaired Student's t-test. g) Mean percentage increase in myelinated axons in trained FIRE+/+ and FIREΔ/Δ mice. n = 3 mice per group in untrained category, 4 trained FIRE+/+ mice and 6 trained FIREΔ/Δ mice. h) Correlation between mean myelinated axons per mm2 and reversal day 1 (RD1) primary errors or average primary errors in FIRE+/+ and FIREΔ/Δ mice. Each data point represents an individual mouse. n = 4 FIRE+/+ and 6 trained FIREΔ/Δ mice./p>1 µm: *P = 0.0263, 2-tailed unpaired Student's t-test. e) Images of mature oligodendrocytes co-expressing OLIG2 (white) and CC1 (magenta) at 6 months of age. Scale bar, 75 µm. f) Mean OLIG2+ CC1+ cells/mm2 ± s.e.m. n = 5 FIRE+/+ and 3 FIREΔ/Δ. Non-significant, P = 0.6400, 2-tailed unpaired Student's t-test. Mean proportion of cells of the oligodendrocyte lineage (OLIG2+) which are mature (CC1+; black) or immature (CC1-; grey) ± s.e.m. n = 5 FIRE+/+ and 3 FIREΔ/Δ. Non-significant, CC1+: P = 0.9938; CC1-: ns P = 0.9938, 1-way ANOVA with Tukey's multiple comparisons test. g) Images of mature oligodendrocytes co-expressing OLIG2 (white) and CC1 (magenta) at 3-4 months of age. Scale bar, 75 µm. h) Mean OLIG2+ CC1+ cells/mm2 ± s.e.m. n = 5 mice/group. Non-significant, P = 0.9825, 2-tailed unpaired Student's t-test. Mean proportion of cells of the oligodendrocyte lineage (OLIG2+) which are mature (CC1+; black) or immature (CC1-; grey) ± s.e.m. n = 5 mice per group. Non-significant, CC1+: P = 0.7076; CC1-: P = 0.7076, 1-way ANOVA with Tukey's multiple comparisons test. i) Images of FIRE+/+ and FIREΔ/Δ mouse corpus callosum at 4.5 months of age indicating onset of demyelination in the latter (magenta asterisks), examples of myelinated axons of medium-large calibre indicated by green asterisks. Scale bars, 5 µm and 1 µm. j) Mean number of myelinated axons/mm2 ± s.e.m. n = 3 mice/group. **P = 0.0042, 2-tailed unpaired Student's t-test. k) Mean myelin thickness per small (<0.6 µm) or medium-large (>0.6 µm) axon diameter bin ± s.e.m. at 3-4 months of age. n = 3 mice/group. <0.6 µm: non-significant, P = 0.8978; >0.6 µm: *P = 0.0491, 2-way ANOVA with Sidak's multiple comparisons test. l) Mean of diameters (0.7290 µm) of demyelinated axons per representative image ± s.e.m. in FIREΔ/Δ mice. n = 3 mice./p>1 µm WT vs. Plp-creERT;Tgfbr1fl/fl **P = 0.0037 and Plp-creERT;Tgfbr1fl/fl vs. Tgfbr1fl/fl ***P = 0.0003, 2-way ANOVA with Sidak's multiple comparisons test. f) Mean number of myelinated axons ± s.e.m./axon diameter bin in 3-month-old FIREΔ/Δ mice following treatment with vehicle control or SRI-011381 hydrochloride from 2 to 3 months of age. n = 3 Vehicle-treated and 4 SRI-treated mice. Non-significant, P = 0.9202, 2-way ANOVA with Sidak's multiple comparisons test. g) Mean inner tongue thickness (µm) ± s.e.m. per axon diameter bin in 3-month-old FIREΔ/Δ mice following treatment with vehicle control or SRI-011381 hydrochloride from 2 to 3 months of age. n = 3 Vehicle-treated and 4 SRI-treated mice. 0.5-1.0 µm: *P = 0.0334, >1 µm: ****P < 0.0001, 2-way ANOVA with Sidak's multiple comparisons test. h) Mean myelin thickness (µm) ± s.e.m./axon diameter bin in 3-month-old FIREΔ/Δ mice following treatment with vehicle control or SRI-011381 hydrochloride from 2 to 3 months of age. n = 3 Vehicle-treated and 4 SRI-treated mice. 0.5-1.0 µm: **P = 0.0027, >1 µm: ****P < 0.0001, 2-way ANOVA with Sidak's multiple comparisons test./p>