Journal: bioRxiv

Article Title: Pervasive environmental chemicals impair oligodendrocyte development

doi: 10.1101/2023.02.10.528042

Figure Lengend Snippet: a, Representative immunohistochemistry images of human cortical organoids treated for 10 days with the flame retardant TDCIPP at 18 µM (approximate IC75 in mPSC-derived oligodendrocytes). Images show all cells (DAPI+, in blue), oligodendrocyte lineage cells (SOX10+, in green), and oligodendrocytes (CC1+, in magenta). b, Quantification of total cell number (DAPI+ per mm2), oligodendrocyte lineage cells (SOX10+ per mm2) and mature oligodendrocytes (SOX10+CC1+ per mm2) in whole cortical organoids. Data are presented as the mean value ± standard deviation from n ≥ 8 biological replicates (individual organoids) indicated by closed circle data points for TDCIPP-treated organoids. p-values were calculated using Student’s two-tailed t test. c, Pie chart showing the number of children ages 3–17 years old from the NHANES 2017–2018 dataset with undetectable and detectable levels of BDCIPP, the urine metabolite of TDCIPP. d, Density plot showing the range and quintiles of urine BDCIPP levels in children ages 3–17 years old from the NHANES 2017–2018 dataset. e, Boxplot showing creatinine-normalized levels of BDCIPP in children 3–17 years of age and adults aged 18 years and older. p-value was calculated using the Kruskal Wallis one-way ANOVA. f, Adjusted odds ratio for the neurodevelopmental outcome: requiring special education or early intervention. Significant odds ratios are highlighted in yellow (BDCIPP Q5 v Q1 OR = 2.7 [95% CI = 1.012–7.407) and gray (Female v Male OR = 0.376 [95% CI = 0.228–0.621]). g, Adjusted odds ratio for the neurodevelopmental outcome: gross motor limitations. Significant odds ratios are highlighted in yellow (BDCIPP Q5 v Q1 OR = 6.0 [95% CI = 1.243–29.426).

Article Snippet: Slides were washed with PBS and incubated overnight with anti-SOX10 (1:200, R&D, AF2864) and anti-APC CC1 (1:200, Millipore, MABC200), followed by labeling with Alexa Fluor-conjugated secondary antibodies (2 μg/mL, Thermo Fisher).

Techniques: Immunohistochemistry, Derivative Assay, Standard Deviation, Two Tailed Test

Journal: iScience

Article Title: Developing a human iPSC-derived three-dimensional myelin spheroid platform for modeling myelin diseases

doi: 10.1016/j.isci.2023.108037

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-CC1 (APC) , Millipore Sigma , Cat# MABC200; RRID: AB_11203645.

Techniques: Recombinant, Software

Journal: The Journal of Cell Biology

Article Title: TDP-43 mediates SREBF2-regulated gene expression required for oligodendrocyte myelination

doi: 10.1083/jcb.201910213

Figure Lengend Snippet: Reduced SREBF2 and its downstream targets in the TDP-43–deleted oligodendrocytes. (A) LDLR and SREBF2 mRNA expression in oligodendrocytes of ctrl and cKO mouse lumbar spinal cord (arrowhead), revealed through combined RNA fluorescent in situ hybridization (RNA-FISH) and GST-P1 IF staining. Images were taken from the ventral gray matter at P21 and P60, at 20× magnification. Scale bar: 20 µm. 3D reconstruction of oligodendrocytes for LDLR (green) and SREBF2 (magenta) mRNA quantification. DAPI (blue). Scale bar: 3 µm. (B) Quantification of SREBF2 (i, ii) and LDLR (iii, iv) puncta in oligodendrocytes of ctrl and cKO mouse lumbar spinal cord, at P21 (i, iii) and P60 (ii, iv). Puncta counts for individual oligodendrocytes (faded circle and triangles), and means (solid circle and triangle) for each animal ( n = 3) plotted, total mean ± SEM derived from all animals. Significance was tested using unpaired t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 3 per genotype, 10 cells were quantified per section, and at least five sections were quantified per animal. (C) Confocal images of colabeling of oligodendrocytes (CC1+, red) and key proteins involved in cholesterol metabolism (SREBF2, left; HMGCS1, middle; and LDLR, right, green) in the white matter of spinal cord sections from 60-d-old mice. Square areas were separated into individual channels, indicating reduction of SREBF2, HMGCS1, and LDLR protein in oligodendrocytes. n = 3 per genotype, six to eight slices per animals were stained and observed. Scale bar: 10 µm. Enlarged images of single cell split into individual channels for SREBF2/HMGCS1/LDLR (green) and CC1 (red). Scale bar: 10 µm. cKO, conditional knockout; ctrl, control.

Article Snippet: The antibodies used in this study were mouse monoclonal APC (Merck; clone CC1; OP80; 1:200), SREBF2 (Abcam; ab30682; 1:400), LDLR (Proteintech; 10785–1-AP; 1:500), and HMGCS1 (Proteintech; 17643–1-AP; 1:500).

Techniques: Expressing, In Situ Hybridization, Staining, Derivative Assay, Knock-Out

Journal: The Journal of Cell Biology

Article Title: TDP-43 mediates SREBF2-regulated gene expression required for oligodendrocyte myelination

doi: 10.1083/jcb.201910213

Figure Lengend Snippet: Reduced expression of cholesterol metabolism in the glia from FTD-TDP patients. (A) Schematic illustration of spinal organoids differentiation from iPSC. (B) Confocal images of TDP-43 (left), HMGCS1 (middle), and HMGCR (right) with mature oligodendrocyte marker (CC1). DAPI nuclear stain in blue. Enlarged images show TDP-43, HMGCS1, and LDLR1 with oligodendrocytes. Scale bar = 50 µm. For magnified images, scale bar = 10 µm. (C) Schematic of post-mortem human FTLD-TDP tissues used for this study. (D) Double IF shows cytoplasmic TDP-43 inclusions (green) associated with white matter oligodendrocytes. Oligodendrocytes with TDP-43 inclusions appear to show a decrease in HMGCR (top, red, arrowheads) and HMGCS1 (middle, red, arrowheads) staining compared with other glial cells (arrows). LDLR colocalizes with TDP-43 inclusions (bottom, red). DAPI nuclear counterstain is blue. White image was taken using 561-nm excitation and corresponds to autofluorescence (autofluo.). Scale bar = 5 µm. DIV, days in vitro; SHH, Sonic Hedgehog; AA, ascorbic acid; WM, white matter.

Article Snippet: The antibodies used in this study were mouse monoclonal APC (Merck; clone CC1; OP80; 1:200), SREBF2 (Abcam; ab30682; 1:400), LDLR (Proteintech; 10785–1-AP; 1:500), and HMGCS1 (Proteintech; 17643–1-AP; 1:500).

Techniques: Expressing, Marker, Staining, In Vitro

Journal: The Journal of Clinical Investigation

Article Title: Axonally derived matrilin-2 induces proinflammatory responses that exacerbate autoimmune neuroinflammation

doi: 10.1172/JCI71385

Figure Lengend Snippet: (A) Colabeling for MATN2 and IBA-1, CC1, or GFAP in the frontal cortices of healthy mice. Scale bar: 50 μm. (B) Labeling for MATN2 on longitudinal spinal cord sections of healthy, acute, and chronic EAE lesions. Scale bar: 12.5 μm. (C) IHC labeling for MATN2 in normal-appearing white matter (NAWM) and early acute and chronic human MS lesions. Acute lesions show lesional extracellular, astrocytic (black arrows), and perivascular expression of MATN2. Scale bar: 200 μm (NAWM); 50 μm (acute); 100 μm (chronic).

Article Snippet: Primary antibodies used were CC1 (Calbiochem, mouse), CD3 (Dako, rabbit), Fn (Chemicon, rabbit), GFAP (Dako, rabbit), IBA-1 (Wako Pure Chemical Industries, rabbit), Lam (Chemicon, rabbit), MATN2 (R&D Systems, goat), MATN2 (generated in-house; ref. 18 ; rabbit), NeuN (Chemicon, mouse), SMI312 (Abcam, mouse).

Techniques: Labeling, Expressing

Journal: Cancer research

Article Title: Age-dependent cellular and behavioral deficits induced by molecularly targeted drugs are reversible

doi: 10.1158/0008-5472.CAN-17-2254

Figure Lengend Snippet: (A) Illustration of the different stages of oligodendrocyte maturation and the subcortical white matter regions used in immunohistochemical analysis. (B) Representative CC1+Olig2+ confocal immunohistochemical images from Paradigm #1 (ED) at P18. (C to H) Quantification of total, (C to D) (Olig2+ cells), mature (E to F) (CC1+Olig2+ cells) and NG2-expressing oligodendrocytes (G to H) in the white matter at different time points. n = 4-5 mice per all groups and per age; one-way ANOVA, Bonferroni post-hoc test for individual comparisons. All data is presented as means ± s.e.m. *p < 0.05, **p < 0.01, ***p < 0.005. Scale bar is 50 microns (μm).

Article Snippet: The following primary antibodies and concentrations were used: mouse CC1, rabbit Olig2, rabbit NG2 chondroitin sulfate proteoglycan, rabbit cleaved (activated) caspase-3 (Casp-3) and guinea pig doublecortin (DCx) antibody were diluted 1:500 (EMD Millipore); chicken antibody glial fibrillary acidic protein (GFAP) was diluted 1:1000 (Abcam); mouse glutamine synthetase (GS), rabbit Sox2, chicken green fluorescent protein (GFP) and rabbit BrdU antibody were diluted 1:500 (Abcam); mouse NeuN antibody was diluted 1:250 (EMD Millipore); and rabbit Ki67 antibody was diluted 1:500 (Vector Labs).

Techniques: Immunohistochemical staining, Expressing

Journal: Cancer research

Article Title: Age-dependent cellular and behavioral deficits induced by molecularly targeted drugs are reversible

doi: 10.1158/0008-5472.CAN-17-2254

Figure Lengend Snippet: (A to D) Quantification of total number of proliferating cells (A to B) (Ki67+) and oligodendrocyte proliferation (C to D) (Ki67+Olig2+) cells for Paradigms #1 and #2. (E to H) Quantification of total apoptotic (E to F) (Casp-3+) and oligodendrocyte apoptotic (G to H) (Casp-3+Olig2+) cells for Paradigms #1 and #2. (I) BrdU pulse protocol for mice randomized to Paradigm #1. (J to K) Quantification of newly generated oligodendrocyte lineage cells (BrdU+Olig2+) and newly generated mature oligodendrocytes (BrdU+CC1+Olig2+) in Paradigm #1 at P30. (L) BrdU pulse protocol for mice randomized to Paradigm #2. (M to N) Quantification of newly generated oligodendrocyte lineage cells (BrdU+Olig2+) and newly generated mature oligodendrocytes (BrdU+CC1+Olig2+) in Paradigm #2 at P30. n = 4-5 mice per all groups and per age; one-way ANOVA, Bonferroni post-hoc test for individual comparisons. All data is presented as means ± s.e.m. *p < 0.05, **p < 0.01, ***p < 0.005.

Article Snippet: The following primary antibodies and concentrations were used: mouse CC1, rabbit Olig2, rabbit NG2 chondroitin sulfate proteoglycan, rabbit cleaved (activated) caspase-3 (Casp-3) and guinea pig doublecortin (DCx) antibody were diluted 1:500 (EMD Millipore); chicken antibody glial fibrillary acidic protein (GFAP) was diluted 1:1000 (Abcam); mouse glutamine synthetase (GS), rabbit Sox2, chicken green fluorescent protein (GFP) and rabbit BrdU antibody were diluted 1:500 (Abcam); mouse NeuN antibody was diluted 1:250 (EMD Millipore); and rabbit Ki67 antibody was diluted 1:500 (Vector Labs).

Techniques: Generated

Journal: Cancer research

Article Title: Age-dependent cellular and behavioral deficits induced by molecularly targeted drugs are reversible

doi: 10.1158/0008-5472.CAN-17-2254

Figure Lengend Snippet: (A) Experimental protocol of vehicle or drug administration, followed by randomization to either receive 12 hours a day of environmental enrichment from P17 – P30 or remain in typical housing environment. BrdU was administered daily from P17 – P20. Behavioral testing was performed between P30 - P32. (B) Quantification of newly generated mature oligodendrocytes (BrdU+CC1+Olig2+) at P30. (C) Quantification of newly generated post-mitotic neurons (BrdU+NeuN+) at P30. (D) The average number of foot slips on the 2 cm and 1 cm inclined beam-walking task. (E) Recognition memory was tested using the novel object recognition memory paradigm with a 12-hour delay between sample and test phase. In the test phase, the percent time spent with the novel object was calculated. For B and C, n = 4 mice per all groups. For D and E, n = 10-15 mice per all groups and per age. A one-way ANOVA, followed by unpaired t-test comparing non-enriched with enrichment. All data is presented as means ± s.e.m. ^p=0.05, *p < 0.05, **p < 0.01, ***p < 0.005.

Article Snippet: The following primary antibodies and concentrations were used: mouse CC1, rabbit Olig2, rabbit NG2 chondroitin sulfate proteoglycan, rabbit cleaved (activated) caspase-3 (Casp-3) and guinea pig doublecortin (DCx) antibody were diluted 1:500 (EMD Millipore); chicken antibody glial fibrillary acidic protein (GFAP) was diluted 1:1000 (Abcam); mouse glutamine synthetase (GS), rabbit Sox2, chicken green fluorescent protein (GFP) and rabbit BrdU antibody were diluted 1:500 (Abcam); mouse NeuN antibody was diluted 1:250 (EMD Millipore); and rabbit Ki67 antibody was diluted 1:500 (Vector Labs).

Techniques: Generated