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. 2019 May 7;8:e44702. doi: 10.7554/eLife.44702

Figure 2. De novo fatty acid synthesis is not required to achieve correct numbers of oligodendrocyte lineage cells during development.

(a) Representative immunostaining of ventral white matter in cross-sectioned spinal cords of P4 and P10 mice identifying proliferating (EdU+) OLs (Olig2+; examples indicated by arrowheads), n = 3 mice for each, CT and cMU. Nuclear marker: DAPI. Scale bar: 50 μm, applying to entire panel. (b) Corresponding graphs to (a) with quantification of percentage of proliferating OLs (EdU+ Olig2+) over total number of OLs (Olig2+) in spinal cord white matter of CT and cMU mice at P4 and P10. Data points represent n = 3 mice for each, CT and cMU; random fields of both dorsal and ventral white matter of 4 sections quantified per animal, with at least 83 Olig2+ cells quantified per section (unpaired two-tailed two sample Student’s t-test; at P4: cMU vs. CT, p=0.6876, t = 0.4326; P10: cMU vs. CT, p=0.5448, t = 0.6608). (c) Representative immunostaining of ventral white matter in cross-sectioned spinal cords from P4, P10, and P14 mice identifying differentiated OLs (CC1+; examples indicated by arrowheads), OLs (Olig2+) and total number of cells (DAPI+), n = 3 mice for each, CT and cMU at P4 and P10, n = 5 mice for each, CT and cMU at P14. Nuclear marker: DAPI. Scale bar: 50 μm, applying to entire panel. (d, e) Corresponding graphs to (c) with quantification of percentage of total OLs (Olig2+) (d) and differentiated OLs (CC1+ Olig2+) (e) over total number of cells (DAPI+), in the spinal cord white matter of CT and cMU mice at P4, P10 and P14. Data points represent n = 3 mice for each, CT and cMU at P4 and P10, and n = 5 mice for each, CT and cMU at P14. Random fields of both dorsal and ventral white matter of at least 3 sections quantified per animal, with at least 83 Olig2+ cells quantified per section (unpaired two-tailed two sample Student’s t-test; % Olig2+/DAPI+ at P4: cMU vs. CT, p=0.8280, t = 0.2319; at P10: cMU vs. CT, p=0.6694, t = 0.46; at P14: cMU vs. CT, p=0.3340, t = 1.028; % CC1+ Olig2+ / DAPI+ at P4: cMU vs. CT, p=0.3405, t = 1.081; at P10: cMU vs. CT, p=0.8490, t = 0.2031; at P14: cMU vs. CT, p=0.2061, t = 1.376). Bars represent mean ±SEM. CT = control, cMU = conditional mutant.

Figure 2.

Figure 2—figure supplement 1. FASN expression in oligodendrocyte progenitors is marginal and dispensable for their early maintenance.

Figure 2—figure supplement 1.

(a) Representative immunostaining of ventral white matter in cross-sections of lumbar spinal cords from P14 CT mice depicting marginal (if any) FASN expression in OPCs (PDGFRα+; examples indicated by arrows and outlining) compared to comparatively high expression in differentiated OLs (CC1+; example indicated by arrowhead). n = 3 examined CT mice. Scale bar: 20 μm, applying to entire panel. (b) Representative immunostainings of ventral white matter in cross-sectioned lumbar spinal cords from P4, P10 and P14 CT and cMU mice, n = 3 mice for each, CT and cMU, at P4 and P10, and n = 4 mice for each, CT and cMU, at P14. Images identify OPCs (PDGFRα+ Olig2+, arrows). Nuclear marker: DAPI. Scale bar: 20 μm, applying to entire panel. (c) Corresponding graphs to (b) with quantification of percentage of OPCs (PDGFRα+ Olig2+) over total number of cells (DAPI+) in the spinal cord white matter of CT and cMU mice at P4, P10 and P14. Data points represent n = 3 mice for each, CT and cMU, at P4 and P10, and n = 4 mice for each, CT and cMU, at P14. Random fields of both dorsal and ventral white matter in at least 3 sections quantified per animal, with at least 121 PDGFRα+ Olig2+ cells quantified per section (unpaired two-tailed two sample Student’s t-test; at P4: cMU vs. CT, p=0.0903, t = 2.223; at P10: cMU vs. CT, p=0.8527, t = 0.198; at P14: cMU vs. CT, p=0.1030, t = 1.922). Bars represent mean ±SEM. CT = control, cMU = conditional mutant.