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Enhanced oligodendrogenesis in the spinal cord gray matter of adult ALS mice. ( a ) Protocol used to trace the fate of NG2 + cells in PDGFαR-CreER;Z/EG ± SOD1 (G93A) mice. Cohorts of NG2 + cells were labeled with EGFP by 4HT injection at P30 or P60 then analyzed at different ages. ( b ) Fluorescence images showing EGFP + NG2 + cell progeny in the ventral horn of the spinal cord in control or SOD1 (G93A) mice 60 days after 4HT administration at P30 (P30+60). Scale bar: 100 μm. ( c – e ) Maps of the location of adult born oligodendrocytes generated from NG2 + cells in the spinal cord of control (blue circles) or SOD1 (G93A) mice (red circles), from P60 to P75 ( c ), from P60 to P90 ( d ), or from P60 to P120 (or end stage for ALS mice) ( e ). Oligodendrocytes (EGFP + CC1 + ) in four randomly sampled lumbar spinal cord sections from each group (from 3–4 mice) are shown. ( f – h ) Graphs showing the density of newly generated EGFP + CC1 + oligodendrocytes in different regions of the adult spinal cord from control (white bars) and SOD1 (G93A) mice (gray bars). Mean + s.e.m. ( n = 9 sections from 3 mice per group) * P < 0.05, ** P < 0.001, *** P < 0.0005, unpaired Student’s t test. i ) Graph showing the overall density of oligodendrocytes (Olig2 + CC1 + ) in different regions of the spinal cord of P120 and end stage SOD1 (G93A) mice. Mean + s.e.m. ( n = 12 sections per group from 4 mice per group) N.S., non-significant, one-way ANOVA with Tukey test.

Journal: Nature neuroscience

Article Title: Degeneration and impaired regeneration of gray matter oligodendrocytes in amyotrophic lateral sclerosis

doi: 10.1038/nn.3357

Figure Lengend Snippet: Enhanced oligodendrogenesis in the spinal cord gray matter of adult ALS mice. ( a ) Protocol used to trace the fate of NG2 + cells in PDGFαR-CreER;Z/EG ± SOD1 (G93A) mice. Cohorts of NG2 + cells were labeled with EGFP by 4HT injection at P30 or P60 then analyzed at different ages. ( b ) Fluorescence images showing EGFP + NG2 + cell progeny in the ventral horn of the spinal cord in control or SOD1 (G93A) mice 60 days after 4HT administration at P30 (P30+60). Scale bar: 100 μm. ( c – e ) Maps of the location of adult born oligodendrocytes generated from NG2 + cells in the spinal cord of control (blue circles) or SOD1 (G93A) mice (red circles), from P60 to P75 ( c ), from P60 to P90 ( d ), or from P60 to P120 (or end stage for ALS mice) ( e ). Oligodendrocytes (EGFP + CC1 + ) in four randomly sampled lumbar spinal cord sections from each group (from 3–4 mice) are shown. ( f – h ) Graphs showing the density of newly generated EGFP + CC1 + oligodendrocytes in different regions of the adult spinal cord from control (white bars) and SOD1 (G93A) mice (gray bars). Mean + s.e.m. ( n = 9 sections from 3 mice per group) * P < 0.05, ** P < 0.001, *** P < 0.0005, unpaired Student’s t test. i ) Graph showing the overall density of oligodendrocytes (Olig2 + CC1 + ) in different regions of the spinal cord of P120 and end stage SOD1 (G93A) mice. Mean + s.e.m. ( n = 12 sections per group from 4 mice per group) N.S., non-significant, one-way ANOVA with Tukey test.

Article Snippet: We used primary antibodies to APC (CC1) (mouse, Calbiochem, 1:50), BrdU (rat, Accurate, 1:500), Caspase-3 (rabbit, Cell Signaling, 1:500), ChAT (goat, Millipore, 1:300), EGFP (goat, Frontier Institute, 1:500), EGFP (rabbit, a gift from Dr. Richard Huganir, Johns Hopkins University, 1:500), GFAP (mouse, NeuroMab, 1:500), GFAP (rabbit, DAKO, 1:1,000), Iba1 (rabbit, Wako, 1:1,000), Ki67 (rabbit, Abcam, 1:500), NG2 (guinea pig, 1:3,600), Olig2 (guinea pig, a gift from Dr. Ben Novitch, UCLA, 1:20,000), Olig2 (rabbit, Millipore, 1:500), PDGFαR (rabbit, a gift from Dr. William Stallcup, Burnham Institute, 1:500), SMI32 (mouse, Covance, 1:1,000), and hSOD1 (rabbit, 1:70).

Techniques: Labeling, Injection, Fluorescence, Control, Generated

Progressive degeneration of oligodendrocytes in the spinal cord ventral gray matter of ALS mice. ( a ) Protocol used to track the fate of early-born oligodendrocytes. Littermate control and SOD1 (G93A) expressing PLP-CreER;ROSA26-EYFP mice received 4HT (2 mg) at P35 and were analyzed at P40, P50, P90 and P120 (or end stage for ALS mice). ( b ) Schematic showing the rationale and possible outcomes of oligodendrocyte fate analysis. ( c,d ) Plot of the number of EYFP + oligodendrocytes (CC1 + Olig2 + ) in ventral gray mater (GM) ( c ) and ventral white matter (WM) ( d ) of PLP-CreER;ROSA26-EYFP ± SOD1 (G93A) mice, expressed relative to the number observed at P35+15 (P50). Note that control group had additional time point (P35+10). Means ± s.e.m. ( n = 9 sections from 3 mice per each time point of each group). * P < 0.05, ** P < 0.001, *** P < 0.0005. Unpaired Student’s t -test was used to compare mean values between control and SOD1 (G93A) mice at each time point. One-way ANOVA with Tukey test was used for age-dependent relative changes in EYFP + oligodendrocytes in each group. ( e ) Confocal images showing the density and morphology of EYFP + (stained with anti-EGFP antibody) CC1 + Olig2 + spinal cord oligodendrocytes (yellow arrows) in ventral gray matter at P120 (control) or at end stage SOD1 (G93A) mice. Scale bars: 20μm.

Journal: Nature neuroscience

Article Title: Degeneration and impaired regeneration of gray matter oligodendrocytes in amyotrophic lateral sclerosis

doi: 10.1038/nn.3357

Figure Lengend Snippet: Progressive degeneration of oligodendrocytes in the spinal cord ventral gray matter of ALS mice. ( a ) Protocol used to track the fate of early-born oligodendrocytes. Littermate control and SOD1 (G93A) expressing PLP-CreER;ROSA26-EYFP mice received 4HT (2 mg) at P35 and were analyzed at P40, P50, P90 and P120 (or end stage for ALS mice). ( b ) Schematic showing the rationale and possible outcomes of oligodendrocyte fate analysis. ( c,d ) Plot of the number of EYFP + oligodendrocytes (CC1 + Olig2 + ) in ventral gray mater (GM) ( c ) and ventral white matter (WM) ( d ) of PLP-CreER;ROSA26-EYFP ± SOD1 (G93A) mice, expressed relative to the number observed at P35+15 (P50). Note that control group had additional time point (P35+10). Means ± s.e.m. ( n = 9 sections from 3 mice per each time point of each group). * P < 0.05, ** P < 0.001, *** P < 0.0005. Unpaired Student’s t -test was used to compare mean values between control and SOD1 (G93A) mice at each time point. One-way ANOVA with Tukey test was used for age-dependent relative changes in EYFP + oligodendrocytes in each group. ( e ) Confocal images showing the density and morphology of EYFP + (stained with anti-EGFP antibody) CC1 + Olig2 + spinal cord oligodendrocytes (yellow arrows) in ventral gray matter at P120 (control) or at end stage SOD1 (G93A) mice. Scale bars: 20μm.

Article Snippet: We used primary antibodies to APC (CC1) (mouse, Calbiochem, 1:50), BrdU (rat, Accurate, 1:500), Caspase-3 (rabbit, Cell Signaling, 1:500), ChAT (goat, Millipore, 1:300), EGFP (goat, Frontier Institute, 1:500), EGFP (rabbit, a gift from Dr. Richard Huganir, Johns Hopkins University, 1:500), GFAP (mouse, NeuroMab, 1:500), GFAP (rabbit, DAKO, 1:1,000), Iba1 (rabbit, Wako, 1:1,000), Ki67 (rabbit, Abcam, 1:500), NG2 (guinea pig, 1:3,600), Olig2 (guinea pig, a gift from Dr. Ben Novitch, UCLA, 1:20,000), Olig2 (rabbit, Millipore, 1:500), PDGFαR (rabbit, a gift from Dr. William Stallcup, Burnham Institute, 1:500), SMI32 (mouse, Covance, 1:1,000), and hSOD1 (rabbit, 1:70).

Techniques: Control, Expressing, Staining

Early disruption of oligodendrocyte structure in the spinal cord of ALS mice. ( a,b ) Confocal images of EGFP + structures in the ventral spinal cord gray matter from control MOBP-EGFP (P120) and MOBP-EGFP;SOD1 (G93A) mice (end stage). Panels in ( b ) show regions highlighted by white squares in ( a ). Arrowheads indicate large EGFP + structures that were Olig2 − and CC1 − . Scale bars: 20 μm. ( c ) Measured volume of Olig2 − EGFP + fragments in each 250,000 μm 3 imaged volume. Mean + s.e.m. ( n = 6 – 9 sections from 3 mice per group) ** P = 1.5 × 10 −5 , *** P = 8 × 10 −6 , unpaired Student’s t -test.

Journal: Nature neuroscience

Article Title: Degeneration and impaired regeneration of gray matter oligodendrocytes in amyotrophic lateral sclerosis

doi: 10.1038/nn.3357

Figure Lengend Snippet: Early disruption of oligodendrocyte structure in the spinal cord of ALS mice. ( a,b ) Confocal images of EGFP + structures in the ventral spinal cord gray matter from control MOBP-EGFP (P120) and MOBP-EGFP;SOD1 (G93A) mice (end stage). Panels in ( b ) show regions highlighted by white squares in ( a ). Arrowheads indicate large EGFP + structures that were Olig2 − and CC1 − . Scale bars: 20 μm. ( c ) Measured volume of Olig2 − EGFP + fragments in each 250,000 μm 3 imaged volume. Mean + s.e.m. ( n = 6 – 9 sections from 3 mice per group) ** P = 1.5 × 10 −5 , *** P = 8 × 10 −6 , unpaired Student’s t -test.

Article Snippet: We used primary antibodies to APC (CC1) (mouse, Calbiochem, 1:50), BrdU (rat, Accurate, 1:500), Caspase-3 (rabbit, Cell Signaling, 1:500), ChAT (goat, Millipore, 1:300), EGFP (goat, Frontier Institute, 1:500), EGFP (rabbit, a gift from Dr. Richard Huganir, Johns Hopkins University, 1:500), GFAP (mouse, NeuroMab, 1:500), GFAP (rabbit, DAKO, 1:1,000), Iba1 (rabbit, Wako, 1:1,000), Ki67 (rabbit, Abcam, 1:500), NG2 (guinea pig, 1:3,600), Olig2 (guinea pig, a gift from Dr. Ben Novitch, UCLA, 1:20,000), Olig2 (rabbit, Millipore, 1:500), PDGFαR (rabbit, a gift from Dr. William Stallcup, Burnham Institute, 1:500), SMI32 (mouse, Covance, 1:1,000), and hSOD1 (rabbit, 1:70).

Techniques: Disruption, Control