{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Alerie Guzman de la Fuente"],"journal":["The Journal of Cell Biology"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-JCBD-201505119"],"attach_to":["JCB"],"legend":["Figure 5: (A-F) Immunostaining with antibodies to VDR together with Sox10 (A), Olig1 (B), MHC class II (C), MOG (D), NeuN (E), and GFAP (F). (G,H) Immunostaining with VDR and MOG shows nuclear localization and higher expression in the plaque border (G) and near absence in a chronic lesion (H). (I) VDR is highly expressed in active plaques and remyelinated plaques (n=5, 1 way ANOVA, Bonferroni post hoc test). (J) Nuclear VDR is expressed in active and shadow plaques to a greater extent than in chronic plaques; cytoplasmic VDR density is simi-lar for all lesion types (n=5, 1 way ANOVA, Bonferroni post hoc test). Mean ± S.E.M are shown. **P<0.01, ***P<0.001. Scale bar 50µm.<br /><br />","Figure 1: (A) Western blot showing expression of NRs in OPCs and mature oligodendrocytes (OLG) (B) CoIP showing the binding between VDR and RXRγ. (C) CoIP of subcellular fractions showing VDR and RXRγ complexes in nucleus and cytoplasm of OPCs and OLG. (D) Quantifi-cation shows no difference in VDR-RXRγ binding in OPCs and OLG (n=3, unpaired Student t-test). (E, F) Duolink immunostaining showing VDR-RXRγ binding (red) in OPCs stained for NG2 (E) and O4 (F). (G) Ethidium bromide is injected into the rat CCP to induce focal demye-lination. Immunostaining of a 21dpl CCP lesion showing colocalization of APC-VDR (H, yellow arrows) and VDR-Olig2 (I, blue arrows). (J) The proportion of Olig2+ cells expressing VDR in the nucleus increases at 14 and 21 dpl being similar to NAWM (n=4, 1 way ANOVA, B","Figure 4: (A) Cerebellar slices were exposed to the vehicle or 2µM ZK159222 for 8DIV: MBP (green) and NFH (red). (B) Treatment with 0.2µM and 2µM ZK159222 impair myelination (n=5, 1 way ANOVA, Dunnett’s post hoc test). (C) Cerebellar slices were demyelinated with lysolecithin (Lyso) and exposed to vehicle (DMSO) and ZK159222 for 8 days. (D) Exposure to 0.2µM and 2µM ZK159222 after demyelination impairs remyelination in a dose-dependent manner (n=3, 2 way ANOVA, Dunnett’s post hoc test). Scale bar 50µm, Mean ± S.E.M are shown.  *P<P0.05, **P<0.01, ***P<0.001","Figure 5: (A-F) Immunostaining with antibodies to VDR together with Sox10 (A), Olig1 (B), MHC class II (C), MOG (D), NeuN (E), and GFAP (F). (G,H) Immunostaining with VDR and MOG shows nuclear localization and higher expression in the plaque border (G) and near absence in a chronic lesion (H). (I) VDR is highly expressed in active plaques and remyelinated plaques (n=5, 1 way ANOVA, Bonferroni post hoc test). (J) Nuclear VDR is expressed in active and shadow plaques to a greater extent than in chronic plaques; cytoplasmic VDR density is simi-lar for all lesion types (n=5, 1 way ANOVA, Bonferroni post hoc test). Mean ± S.E.M are shown. **P<0.01, ***P<0.001. Scale bar 50µm.","Supplementary figure 1: (A) OPCs were treated for 24h with vehicle, 50nM 9cRA or 50nM 9cRA plus ZK159222. (B) Treatment of OPCs with 9cRA and ZK15922 shows that Zk159222 restores 9cRA differentiation to control levels (n=3, 1 way ANOVA, Bonferroni post hoc test). Scale bar 50µm. Mean ± S.E.M. are shown. ***P<0.001.","Supplementary Figure 2: (A) Representative images of OPCs treated with vehicle or 0.1µM VitD and stained for MBP and Olig2. (B) Vitamin D does not increase OPC differentiation (n=3, 1 way ANOVA). (C, D) Vitamin D does not alter OPC proliferation (n=4, 1 way ANOVA). (E) Vitamin D does not cause cell death as measured by PI staining (n=3, Kruskal Wallis). (F) OPCs were treated with vehicle, 9cRA or VitD together with 9cRA for 24h. (G) Co-treatment with vitamin D and 9cRA has no additional effect on OPC differentiation (n=3, 1 way ANOVA, Bonferroni post hoc test). Scale bar 50µm. Mean ± S.E.M. are shown.","Figure 4: (A) Cerebellar slices were exposed to the vehicle or 2µM ZK159222 for 8DIV: MBP (green) and NFH (red). (B) Treatment with 0.2µM and 2µM ZK159222 impair myelination (n=5, 1 way ANOVA, Dunnett’s post hoc test). (C) Cerebellar slices were demyelinated with lysolecithin (Lyso) and exposed to vehicle (DMSO) and ZK159222 for 8 days. (D) Exposure to 0.2µM and 2µM ZK159222 after demyelination impairs remyelination in a dose-dependent manner (n=3, 2 way ANOVA, Dunnett’s post hoc test). Scale bar 50µm, Mean ± S.E.M are shown.  *P","Figure 4: (A) Cerebellar slices were exposed to the vehicle or 2µM ZK159222 for 8DIV: MBP (green) and NFH (red). (B) Treatment with 0.2µM and 2µM ZK159222 impairs myelination (n=5, 1 way ANOVA, Dunnett’s post hoc test). (C) Cerebellar slices were demyelinated with lysolecithin (Lyso) and exposed to vehicle (DMSO) and ZK159222 for 8 days. (D) Exposure to 0.2µM and 2µM ZK159222 after demyelination impairs remyelination in a dose-dependent manner (n=3, 2 way ANOVA, Dunnett’s post hoc test). Scale bar 50µm, Mean ± S.E.M are shown.  *P<P0.05, **P<0.01, ***P<0.001","Figure 3: (A) OPCs treated with vehicle or 0.1µM Vitamin D and stained for MBP and Olig2.  (B) Vitamin D increases OPC differentiation (n=4, 1 way ANOVA, Dunnett’s post hoc test). (C, D) Western blots demonstrating increased MBP after 48h of VitD treatment (n=5, 1 way ANOVA, Dunnett’s post hoc test). (E, F) Vehicle and 0.1µM VitD treated OPCs stained for Ki67 and Olig2. Treatment with VitD decreases the number of Olig2+ cells proliferating (n=6, 1 way ANOVA Dunnett’s post hoc test). (G) Vitamin D treatment did not alter OPC cell death as measured by PI (n=2, 1 way ANOVA). (H,I) Treatment with both VitD and ZK159222 in OPCs derived from MGCs exposed to control (H) (n=7, 1 way ANOVA, Dunnett’s post hoc test) and charcoal-stripped serum (I) (n=3, 1 way ANOVA, Dunnett’s posthoc test) show that","Figure 2: (A) OPCs treated with vehicle or 0.2µM ZK159222 were stained with antibodies to Ki67 and Olig2. (B) Treatment with ZK15922 for 24h increases OPC proliferation (n=5, 1 way ANOVA, Dunnett’s posthoc test) (C) OPCs treated with vehicle or 0.2µM ZK159222 stained for MBP and Olig2. (D) Treatment with ZK159222 for 24h impairs OPC differentiation (n=5, 1 way ANOVA, Dunnett’s posthoc test). (E, F) Treatment with ZK159222 for 48h decreases MBP protein levels (n=3, 1 way ANOVA, Dunnett’s posthoc test). (G) Treatment with ZK159222 does not affect cell death quantified by propidium iodide staining (n=4, 1 way ANOVA). (H-L) Purified OPCs were treated with non-targeting siRNA or VDR siRNA for 48h and stained with antibodies to MBP(H), Ki67 (I) and Olig2. (J) Western blot showing that VDR protei"],"repository":["bioimages"],"figure_sub":["Image 632949 (Figure 3 - C)","Image 632860 (Figure 2 - C)","Figure S2 - G","Figure S2 - D","Image 632883 (Figure 6 - A)","Image 632951 (Figure 1 - B)","Image 632911 (Figure 5 - D)","Figure S2 - A","Image 632958 (Figure 1 - A)","Image 632866 (Figure 2 - I)","Image 632869 (Figure 4 - C)","Image 632983 (Figure S2 - A)","Image 632978 (Figure S2 - D)","Image 632955 (Figure 1 - A)","Image 632877 (Figure 5 - E)","Image 632982 (Figure S2 - G)","Figure 1 - H and I","Image 632975 (Figure 3 - E)","Figure S2","Image 632880 (Figure 5 - H)","Image 632966 (Figure 3 - A)","Image 632969 (Figure 2 - A)","Image 632861 (Figure 2 - E)","Image 632963 (Figure 1 - C)","Image 632976 (Figure S2 - A)","Image 632965 (Figure 1 - F)","Image 632863 (Figure 2 - H)","Image 632956 (Figure 1 - A)","Figure 1 - F","Image 632910 (Figure 5 - A)","Figure 1 - A","Image 632862 (Figure 2 - E)","Figure 1 - E","Image 632950 (Figure 3 - A)","Image 632960 (Figure 1 - A)","Figure 1 - C","Figure 1 - B","Image 632970 (Figure 2 - A)","Image 632909 (Figure 4 - A )","Figure 2 - H","Figure 2 - E","Figure 2 - I","Figure 2 - J","Image 632979 (Figure S2 - D)","Figure 2 - C","Image 632878 (Figure 5 - F)","Figure 2 - A","Image 632980 (Figure S2 - G)","Figure 3 - E","Image 632974 (Figure 3 - E)","Image 632954 (Figure 1 - A)","Image 632879 (Figure 5 - G)","Image 632868 (Figure 2 - J)","Figure 3 - A","Image 632962 (Figure 1 - C)","Figure 3 - C","Image 632864 (Figure 2 - H)","Figure 4 - A","Image 632948 (Figure 3 - C)","Image 632952 (Figure 1 - B)","Image 632972 (Figure 1 - A)","Image 632865 (Figure 2 - I)","Image 632959 (Figure 1 - A)","Image 632961 (Figure 1 - B)","Image 632971 (Figure 1 - H and I)","Figure 4 - C","Image 632859 (Figure 2 - C)","Figure 5 - A","Image 632875 (Figure 5 - C)","Figure 5 - F","Figure 5 - G","Image 632881 (Figure 6 - A)","Image 632981 (Figure S2 - G)","Figure 5 - H","Figure 5 - B","Figure 5 - C","Image 632964 (Figure 1 - E)","Figure 5 - D","Figure 5 - E","Figure 5","Figure 4","Figure 6","Image 632870 (Figure 4 - C)","Image 632882 (Figure 6 - A)","Image 632908 (Figure 4 - A )","Image 632953 (Figure 1 - A)","Figure 6 - A","Figure 1","Image 632947 (Figure 2 - J)","Image 632874 (Figure 5 - B)","Figure 3","Figure 2"],"pubmed_authors":["Hilary J. Lewis","Robin J.M. Franklin","Peter van Wijngaarden","Jeffrey K. Huang","Charles ffrench-Constant","Christophe Kerninon","Alerie Guzman de la Fuente","Clare A. Jones","Oihana Errea","Ginez A. Gonzalez","Andrew A. Jarjour","Brahim Nait-Oumesmar","Chao Zhao"],"additional_accession":[]},"is_claimable":false,"name":"Vitamin D receptor–retinoid X receptor heterodimer signaling regulates oligodendrocyte progenitor cell differentiation","description":"The mechanisms regulating differentiation of oligodendrocyte (OLG) progenitor cells (OPCs) into mature OLGs are key to understanding myelination and remyelination. Signaling via the retinoid X receptor γ (RXR-γ) has been shown to be a positive regulator of OPC differentiation. However, the nuclear receptor (NR) binding partner of RXR-γ has not been established. In this study we show that RXR-γ binds to several NRs in OPCs and OLGs, one of which is vitamin D receptor (VDR). Using pharmacological and knockdown approaches we show that RXR–VDR signaling induces OPC differentiation and that VDR agonist vitamin D enhances OPC differentiation. We also show expression of VDR in OLG lineage cells in multiple sclerosis. Our data reveal a role for vitamin D in the regenerative component of demyelinat","dates":{"release":"2015-12-07T11:27:59Z","modification":"2018-11-29T11:27:59Z","creation":"2018-11-29T11:27:59Z"},"accession":"S-JCBD-201505119","cross_references":{"doi":["10.1083/jcb.201505119"]}}