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

Figure 7. Generation of tamoxifen-inducible PdgfrαCreERT2:FASN-floxed conditionally mutant mice.

(a) Tamoxifen-mediated nuclear translocation of Pdgfrα-driven CreERT2 induces conditional Fasn allele inactivation (i-cMU) and expression of yellow fluorescent protein (YFP). (b) Timeline of de-/re-myelination experiments: Focal demyelination of the ventral spinal cord white matter was induced by injection of lysolecithin 4 weeks post-tamoxifen administration. The tissue was analysed 7, 10, 14, 21 and 56 days post-lysolecithin injection (dpl). (c) Schematic of lysolecithin-induced demyelinated focal lesion in the ventral funiculus of the thoracic spinal cord (level T12). Square insert indicates the area where immunostaining images were acquired. (d) Representative immunostainings of lesioned areas in cross-sectioned thoracic spinal cords from CT and i-cMU mice 7 dpl, n = 3 mice analyzed for each, CT and i-cMU. Note the prominent FASN expression in the cytoplasm of recombined aOPC-derived differentiated OLs (CC1+ YFP+ FASN+, examples indicated by arrowheads) in CT, but not in i-cMU (CC1+ YFP+ FASN-, examples indicated by dotted lines) lesions. Nuclear marker: DAPI. Scale bar: 20 μm, applies to entire panel. CT = control, i-cMU = inducible conditional mutant, dpl = days post-lysolecithin injection.

Figure 7.

Figure 7—figure supplement 1. Recombination efficiency in spinal cords of PdgfrαCreERT2:Rosa26-loxPstoploxP-YFP control animals induced with tamoxifen.

Figure 7—figure supplement 1.

(a) Exemplary immunostaining of cross-sectioned thoracic spinal cord white matter from 5 weeks post tamoxifen, n = 3 CT mice. Cre-dependent expression of the reporter Rosa26-loxPstoploxP-YFP allele is detectable in aOPCs (PDGFRα+ YFP+, example indicated by arrow), whereas other aOPCs remain unrecombined (PDGFRα+ YFP-, examples indicated by arrowheads). Nuclear marker: DAPI. Scale bar: 20 μm, applying to entire panel. (b) Corresponding graph to (a) with quantification of percentage of recombined aOPCs (PDGFRα+ YFP+) over total aOPCs (PDGFRα+). Data points represent n = 3 CT mice; 3 spinal cord sections were quantified per animal, with at least 36 PDGFRα+ YFP+ cells quantified per animal. Bar represents mean ± SEM.