Fig. 1

A novel inducible myelinating glia-specific CST cKO mouse model recapitulates AD-like adult-onset CNS-specific sulfatide loss without affecting oligodendrocyte/myelin homeostasis. (A) Tamoxifen injection and tissue harvest/analysis protocol. CST Cre⁻ and CST Cre⁺ mice were injected with tamoxifen at 3–4 mo of age and sacrificed at 4.5 and 9 mo post-injection, at 7.5 and 12 mo of age, respectively. (B) RNA extracted from the cerebrum (CRM) and spinal cord (SC) of CST Cre⁻ and CST Cre⁺ mice was assessed using the NanoString nCounter mouse Neuroinflammation Panel, linear mRNA counts for the Cst (Gal3st1) gene are shown. NanoString nSolver recommended test (heteroscedastic Welch’s t-Test), n = 3–4. (C) Lipid extracts from CRM, SC, and sciatic nerve (SN) of CST Cre⁻ and CST Cre⁺ mice were assessed by shotgun lipidomics; total sulfatide levels are shown as nmol/mg of total protein. See Fig. S2B-F for additional lipid classes. Two-way ANOVA with Bonferroni posthoc test for multiple comparisons, n = 4–8. (D) Heatmap displaying log2 fold changes of oligodendrocyte/myelin function-related genes included in the Nanotring panel (except Gal3st1, which is shown separately) from CRM, SC of CST cKO mice, as well as from 1 and 2 mo old CST KO mice for comparison, relative to their respective controls. Heteroscedastic Welch’s t-Test, n = 3–4. P-value is displayed within each heat map cell (*p < 0.05). (E) Oligodendrocyte pathway scores were obtained using nSolver 4.0 Advanced Analysis (NanoString Technologies). Two-way ANOVA with Bonferroni posthoc test for multiple comparisons, n = 3–4. (F) CRM and SC NP40 homogenate supernatants were assessed by Western blot using antibodies against myelin-associated glycoprotein (MAG) and 2′,3′-Cyclic nucleotide 3′-phosphodiesterase (CNP). Two-tailed unpaired t-Test, n = 3–4. *p < 0.05, **p < 0.01, ***p < 0.001. Data represent the mean ± S.E.M