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. 2022 Mar 21;11:e75230. doi: 10.7554/eLife.75230

Figure 2. Early deletion of TDP-43 in premyelinating oligodendrocytes leads to seizure development, premature death, and increased oligodendrocyte turnover.

(A) Diagram of oligodendrocyte development showing Mobp-iCre induces Tardbp deletion at the premyelinating oligodendrocyte stage whereas Mog-iCre targets myelinating oligodendrocytes. (B) Kaplan–Meier survival curve for Mobp-TDP43 (orange) and Mog-TDP43 (blue) mouse lines. Dotted line indicates the 95% confidence interval. Log-rank (Mantel–Cox) statistical test showed significance at p value <0.0001 (n = 58, 33, 40, and 32). Red arrowhead points to ~P110 when the animals first exhibit spontaneous seizures. (C) ASPA+ oligodendrocyte staining in the motor cortex of Mobp-TDP43 and MOG-TDP43 mouse lines at P30, P90, and P180. (D) Quantification of ASPA+ oligodendrocyte density shows that there is no statistical difference between the samples at any given timepoint (one-way ANOVA with Tukey’s multiple comparisons test, n = 3, n.s. p value >0.05). (E) Immunostaining for Ki-67 and NG2 to identify proliferating oligodendrocyte precursor cells (OPCs) in Mobp-TDP43 and Mog-TDP43 at P90. (F) Quantification of Ki-67+NG2+ proliferating OPC density shows that it is statistically significantly increased in Mobp-TDP43 KO whereas Mog-TDP43 KO shows a trend toward an increased density (one-way ANOVA with Tukey’s multiple comparisons test, n = 4, ***p value <0.001, n.s. p value = 0.3776).

Figure 2.

Figure 2—figure supplement 1. Quantification of Mog-iCre recombination efficiency.

Figure 2—figure supplement 1.

(A) Maximum intensity z-projection confocal images of the motor cortex from Mog-iCre × RCE mice at P90 and P180 immunostained with GFP and ASPA. (B) Quantification of % recombination (EGFP+ASAP+/ASPA+) and % ASPA+/EGFP+ to show recombination efficiency and specificity in P90 and P180 mouse cortex. n = 3.
Figure 2—figure supplement 2. Characterization of Mobp-iCre and Mobp-iCreERT2 mouse lines.

Figure 2—figure supplement 2.

(A) Schematic of tamoxifen injection in different Mobp-iCreER mouse lines at P35. (B) Representative images of ASPA (oligodendrocyte marker) and NG2 (oligodendrocyte precursor cell [OPC] marker) from Line #10 without tamoxifen and Line #8,16 with and without tamoxifen injections. (C) Quantification of % recombination (EGFP+ASAP+/ASPA+) and % ASPA+/EGFP+ to show recombination efficiency and specificity in Lines #10 and #8,16 in cortex, corpus callosum (CC), spinal cord gray matter (SC GM), and spinal cord white matter (SC WM).
Figure 2—figure supplement 3. Mobp-TDP43 and Mog-TDP43 mouse lines exhibit differences in response to loss of TDP-43.

Figure 2—figure supplement 3.

(A) Mobp and Mog expression in oligodendrocyte lineage cells (adapted from https://greenberg.hms.harvard.edu/gene-database/ Hrvatin et al., 2018). (B) Mobp and Mog expression in the lineage of oligodendrocytes (adapted from http://linnarssonlab.org/oligodendrocytes/ Marques et al., 2016).
Figure 2—figure supplement 4. Differential gene expression between premyelinating (orange) and myelinating (blue) oligodendrocytes (OL) (reproduced from the raw single-cell RNA-Seq dataset in Hrvatin et al., 2018).

Figure 2—figure supplement 4.

Top 16 differentially expressed genes are shown. All genes are statistically significantly differentially expressed (q value <0.01).
Figure 2—figure supplement 5. Mobp-TDP43 KO animals exhibit a higher oligodendrocyte turnover rate and increased astrogliosis.

Figure 2—figure supplement 5.

(A) In situ hybridization for lncOL1 (9630013A20Rik) shows increased number of premyelinating oligodendrocytes in P150 Mobp-TDP43 KO compared to the Control. (B) Immunostaining for GFAP in the motor cortex at P180 in Mobp-TDP43 and Mog-TDP43 mouse lines. Scale bar = 500 μm. (C) Quantification of mean GFAP intensity (arbitrary unit; a.u.) in the motor cortex. One-way ANOVA with Tukey’s multiple comparisons test, ***p value <0.001, n.s. p value >0.05.