Journal: Frontiers in Neuroscience

Article Title: Disease-associated oligodendrocyte signatures are spatiotemporally dysregulated in spinocerebellar ataxia type 3

doi: 10.3389/fnins.2023.1118429

Figure Lengend Snippet: Onset of behavioral deficits in Q84 mice associates with mature oligodendrocyte transcript dysregulation. (A) Schematic of the recording timepoints for four phenotypic characterizations of Q84 mice. Tissue was collected for qPCR analysis at 3-, 4-, 8-, and 16-weeks-old. Q84; Mobp -eGFP tissue was collected for immunohistochemical analysis at 4-, 8-, and 16-weeks-old. Recordings of panel (B) weight, (C) total locomotor activity (x-y beam breaks), and (D) rearing activity (z beam breaks) in hemizygous (Q84/WT) and homozygous (Q84/Q84) Q84 mice relative to their WT littermates (WT/WT) at 2-, 3-, 4-, 8-, and 14-weeks-old ( n = 16–20 mice/genotype. Mixed-effects analysis with a post-hoc Tukey’s multiple comparison test was performed for behavioral analyses * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; no asterisk, not significant. Red asterisks indicate comparison of Q84/Q84 to WT/WT. (E,F) QPCR analysis of oligodendrocyte maturation markers ( Plp1 , Mal , Mobp ) in Q84 mice in the (E) brainstem, (F) spinal cord, (G) cerebellum, and (H) forebrain. Each data points represents an independent mouse per genotype. For qPCR analysis, unpaired Student’s t -tests or Mann–Whitney tests were performed, depending on normality distribution as determined by Shapiro–Wilk tests: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: TaqMan primers used include the following: Beta Actin (ThermoFisher; Cat # Mm02619580; Burlington, MA, United States), Plp1 (ThermoFisher; Cat #Mm01297210_m1; Burlington, MA, United States), Ugt8a (ThermoFisher; Cat #Mm00495930_m1; Burlington, MA, United States), Mobp (ThermoFisher; Cat #Mm02745649_m1; Burlington, MA, United States), and Mal (ThermoFisher; Cat #Mm01339780_m1; Burlington, MA, United States).

Techniques: Immunohistochemical staining, Activity Assay, MANN-WHITNEY

Journal: Frontiers in Neuroscience

Article Title: Disease-associated oligodendrocyte signatures are spatiotemporally dysregulated in spinocerebellar ataxia type 3

doi: 10.3389/fnins.2023.1118429

Figure Lengend Snippet: Mature oligodendrocytes are reduced in a spatiotemporal pattern that corresponds to disease progression. (A) Mouse cross between Q84 SCA3 mice and Mobp -eGFP reporter mice. (B) Depiction of where images were taken for histologically assessed regions. (C) Representative images of Mobp -eGFP+ mature oligodendrocytes in the pons, DCN, CST, CC, cortex, and striatum at 4 weeks (left) and 16 weeks of age (right). Scale bar 50 μm. (D) Quantification of mature oligodendrocytes in the pons, DCN, CST, CC, cortex, and striatum of Q84; Mobp -eGFP+ mice at 4 (left), 8 (middle), and 16 weeks of age (right) ( n = 4–6 mice/genotype, 3 images/mouse). One-way ANOVA with a post-hoc Tukey’s multiple comparisons test was performed: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: TaqMan primers used include the following: Beta Actin (ThermoFisher; Cat # Mm02619580; Burlington, MA, United States), Plp1 (ThermoFisher; Cat #Mm01297210_m1; Burlington, MA, United States), Ugt8a (ThermoFisher; Cat #Mm00495930_m1; Burlington, MA, United States), Mobp (ThermoFisher; Cat #Mm02745649_m1; Burlington, MA, United States), and Mal (ThermoFisher; Cat #Mm01339780_m1; Burlington, MA, United States).

Techniques:

Journal: Bioengineered

Article Title: Four specific biomarkers associated with the progression of glioblastoma multiforme in older adults identified using weighted gene co-expression network analysis

doi: 10.1080/21655979.2021.1975980

Figure Lengend Snippet: Detail of hub genes in each significant module

Article Snippet: The primary antibodies including ERMN (1:200; Cat No. 66,605-1-Ig, Proteintech, Wuhan, China), MOBP (1:100; Cat No.12472-1-AP, Proteintech, Wuhan, China), PLP1 (1:100; Cat No. A20009, Abconal, Wuhan, China) and OPALIN (1:300; Cat. ab121425, Abcam, China), were added to the sections and incubated overnight at 4°C, followed by incubation with anti-mouse (1:2000; cat. no. BM3895) and anti-rabbit (1:2000; cat. no. BM3894) horseradish peroxidase-conjugated goat secondary antibodies (Boster, Wuhan, China) after washing three times with PBS.

Techniques:

Journal: Bioengineered

Article Title: Four specific biomarkers associated with the progression of glioblastoma multiforme in older adults identified using weighted gene co-expression network analysis

doi: 10.1080/21655979.2021.1975980

Figure Lengend Snippet: ERMN, MOBP, PLP1 and OPALIN were highly expressed in the GBM tissues provided by the older patients with lower KPS scores . (a) IHC stain determined the expression of ERMN, MOBP, PLP1 and OPALIN in GBM tissues provided by the older patients with low and high KPS scores. (b) ROC analysis was performed to determine the diagnostic value of ERMN, MOBP, PLP1 and OPALIN to distinguish the GBM tissues provided by the older patients with low and high KPS scores

Article Snippet: The primary antibodies including ERMN (1:200; Cat No. 66,605-1-Ig, Proteintech, Wuhan, China), MOBP (1:100; Cat No.12472-1-AP, Proteintech, Wuhan, China), PLP1 (1:100; Cat No. A20009, Abconal, Wuhan, China) and OPALIN (1:300; Cat. ab121425, Abcam, China), were added to the sections and incubated overnight at 4°C, followed by incubation with anti-mouse (1:2000; cat. no. BM3895) and anti-rabbit (1:2000; cat. no. BM3894) horseradish peroxidase-conjugated goat secondary antibodies (Boster, Wuhan, China) after washing three times with PBS.

Techniques: Staining, Expressing, Diagnostic Assay

Journal: Bioengineered

Article Title: Four specific biomarkers associated with the progression of glioblastoma multiforme in older adults identified using weighted gene co-expression network analysis

doi: 10.1080/21655979.2021.1975980

Figure Lengend Snippet: Detail IHC score of ERMN, MOBP, PLP1 and OPALIN in GBM tissues provided by the elder patients with low KPS score and high KPS score

Article Snippet: The primary antibodies including ERMN (1:200; Cat No. 66,605-1-Ig, Proteintech, Wuhan, China), MOBP (1:100; Cat No.12472-1-AP, Proteintech, Wuhan, China), PLP1 (1:100; Cat No. A20009, Abconal, Wuhan, China) and OPALIN (1:300; Cat. ab121425, Abcam, China), were added to the sections and incubated overnight at 4°C, followed by incubation with anti-mouse (1:2000; cat. no. BM3895) and anti-rabbit (1:2000; cat. no. BM3894) horseradish peroxidase-conjugated goat secondary antibodies (Boster, Wuhan, China) after washing three times with PBS.

Techniques:

Journal: bioRxiv

Article Title: BrainPalmSeq: A curated RNA-seq database of palmitoylating and de-palmitoylating enzyme expression in the mouse brain

doi: 10.1101/2021.11.23.468539

Figure Lengend Snippet: (A) Graph of expression data for Zdhhc9 extracted from BrainPalmSeq. Original data from oligodendrocyte scRNAseq study by Marques et al. Expression units: mean log2(counts per 10,000 + 1). (B) Diagram illustrating workflow to generate a list of oligodendrocyte enriched palmitoylation substrates, GO annotated for myelin sheath for experimental validation. (C) STRING diagram of myelin sheath annotated palmitoylation substrates. (D) Western blot following Acyl-RAC palmitoylation assay in HEK293 cells to identify palmitoylated and unpalmitoylated fractions of FLAG-MOBP either without or with co-transfection of FLAG-GOLGA7 and HA-ZDHHC9. Input = unprocessed protein lysate. NSB control = non-specific binding of unpalmitoylated protein to sepharose resin in control pipeline. Palm fraction = palmitoylated protein (bound to sepharose resin). Unpalm fraction = unpalmitoylated protein (did not bind to sepharose resin). (E-F) As in (D) but for FLAG-PLP1 (E) or FLAG-CNP (F). (G) Graphs quantifying the ratio of palmitoylated to unpalmitoylated protein either with or without co-transfections with FLAG-GOLGA7 and HA-ZDHHC9. n = 4-6 HEK cell cultures per condition. Statistics shown for palmitoylated fraction. Two-way ANOVA; Šídák’s post hoc ; mean ± SEM. MOBP: p = 0.0004, 95% CI −0.6594 to −0.2266; PLP1: p = 0.0046, 95% CI −0.1660 to −0.03011; CNP: p = 0.6981, 95% CI - 0.3274 to 0.1742.

Article Snippet: Human FLAG-MOBP (CAT#: RC223946), FLAG-PLP1 (CAT#: RC218616) and FLAG-CNP (CAT#: RC207038) were acquired from Origene, Maryland, USA.

Techniques: Expressing, Western Blot, Cotransfection, Binding Assay, Transfection

Journal: The American Journal of Pathology

Article Title: Modulation of Sox10, HIF-1α, Survivin, and YAP by Minocycline in the Treatment of Neurodevelopmental Handicaps following Hypoxic Insult

doi: 10.1016/j.ajpath.2015.05.016

Figure Lengend Snippet: Western blot analysis of postnatal day 20 (P20) isolated SVZ tissues harvested from normoxia-reared nursing pups treated with either plain water, 5% sucrose (vehicle), or minocycline and reduced oxygen–reared nursing pups treated with either plain water, 5% sucrose (vehicle), or minocycline. Minocycline treatment of hypoxic-reared nursing C57BL/6 pups elicits statistically significant increases in Sox10 (A), HIF-1α (B), EPO (C), BDNF (D), phospho-AKT (E), cleaved notch1 (F), YAP (G), survivin (H), GPR124 (I), CD31 (J), PCNA (K), MAG (M), PLP (N), MOBP (O), and synapsin (P), and decreases in CCASP3(L) compared to vehicle-only and normoxia-reared pups. Data are expressed as means ± SD. n = 3. ∗P < 0.05, ∗∗P < 0.005. CCASP3, cleaved caspase 3; EPO, erythropoietin; GPR, G-protein coupled receptor; H2O, water-treated; HIF, hypoxia-inducible factor; HX, hypoxic; MAG, myelin associated glycoprotein; MC, minocycline-treated; MOBP, myelin-associated oligodendrocyte basic protein; NX, normoxic; PCNA, proliferating cell nuclear antigen; PLP, proteolipid protein; SC, sucrose (vehicle); SVZ, subventricular zone; YAP, yes-associated protein 1.

Article Snippet: Antibody directed against Sox10 was purchased from Abgent (San Diego, CA); antibodies directed against HIF-1α, HIF-2α, PLP, and GPR124 were purchased from Novus Biologicals (Littleton, CO); antibodies directed against proliferating cell nuclear antigen (PCNA), cleaved Notch1, cleaved caspase 3, synapsin, Sox2, survivin, and YAP were purchased from Cell Signaling Technology (Danvers, MA); antibodies directed against MAG and MOBP were purchased from Santa Cruz Biotechnology (Dallas, TX); antibodies directed against β-actin, Ki-67, and glial fibrillary acidic protein (GFAP) were purchased from Abcam (Cambridge, MA); antibodies directed against nestin were purchased from Millipore (Darmstadt, Germany).

Techniques: Western Blot, Isolation

Journal: The American Journal of Pathology

Article Title: Modulation of Sox10, HIF-1α, Survivin, and YAP by Minocycline in the Treatment of Neurodevelopmental Handicaps following Hypoxic Insult

doi: 10.1016/j.ajpath.2015.05.016

Figure Lengend Snippet: Western blot analysis of postnatal day 70 (P70) isolated SVZ tissues harvested from reduced oxygen–reared nursing pups treated with either 5% sucrose (vehicle), or minocycline. A–G: Similar to the changes noted in SVZ tissues at P20 and P40, Sox10 (A), HIF-1α (B), PCNA (C), cleaved notch1 (D), CCASPAS3 (E), GPR124 (G), and PLP (G) are still elevated in the minocycline-treated pups compared to the vehicle-only treated pups. Changes noted in the cerebral cortex mirror those noted in the SVZ tissues at P20 and P40. Specifically, significant increases are noted in Sox10 (H), cleaved notch1 (I), HIF-1α (J), HIF-2α (K), PCNA (L), MAG (N), MOBP (O), PLP (P), and synapsin (Q) and decreases in CCASPAS3 (M) in minocycline treated pups. Data are expressed as means ± SD. n = 3. ∗P < 0.05, ∗∗P < 0.005. CCASPAS3, cleaved caspase 3; CT, control; GPR, G-protein coupled receptor; HIF, hypoxia-inducible factor; IB, immunoblot; MAG, myelin associated glycoprotein; MC, minocycline-treated; MOBP, myelin-associated oligodendrocyte basic protein; PCNA, proliferating cell nuclear antigen; PLP, proteolipid protein; SVZ, subventricular zone.

Article Snippet: Antibody directed against Sox10 was purchased from Abgent (San Diego, CA); antibodies directed against HIF-1α, HIF-2α, PLP, and GPR124 were purchased from Novus Biologicals (Littleton, CO); antibodies directed against proliferating cell nuclear antigen (PCNA), cleaved Notch1, cleaved caspase 3, synapsin, Sox2, survivin, and YAP were purchased from Cell Signaling Technology (Danvers, MA); antibodies directed against MAG and MOBP were purchased from Santa Cruz Biotechnology (Dallas, TX); antibodies directed against β-actin, Ki-67, and glial fibrillary acidic protein (GFAP) were purchased from Abcam (Cambridge, MA); antibodies directed against nestin were purchased from Millipore (Darmstadt, Germany).

Techniques: Western Blot, Isolation, Control

Journal: Aging (Albany NY)

Article Title: Amyotrophic lateral sclerosis, gene deregulation in the anterior horn of the spinal cord and frontal cortex area 8: implications in frontotemporal lobar degeneration

doi: 10.18632/aging.101195

Figure Lengend Snippet: Main significant clusters of altered genes in frontal cortex of ALS samples

Article Snippet: Myelin-Associated Oligodendrocyte Basic Protein , MOBP , Hs01094434_m1.

Techniques: Binding Assay, Activity Assay, Membrane, Transmission Assay, Translocation Assay

Journal: Aging (Albany NY)

Article Title: Amyotrophic lateral sclerosis, gene deregulation in the anterior horn of the spinal cord and frontal cortex area 8: implications in frontotemporal lobar degeneration

doi: 10.18632/aging.101195

Figure Lengend Snippet: Genes, gene symbols and TaqMan probes used for the study of gene expression in the anterior horn of the spinal cord and frontal cortex area 8 in ALS cases and controls including probes for normalization ( AARS , GUS-β , HPRT-1 and XPNPEP-1 )

Article Snippet: Myelin-Associated Oligodendrocyte Basic Protein , MOBP , Hs01094434_m1.

Techniques: Gene Expression, Immunopeptidomics

Journal: bioRxiv

Article Title: Atxn2 -CAG100-knock-in affects mouse lifespan and vestibulo-cerebellar function via neural disconnection

doi: 10.1101/333443

Figure Lengend Snippet: Downregulations of mRNA levels were observed for several factors involved in axon myelination, where specific and sensitive antibodies were unavailable. (A) Among oligodendrocytic factors, this included Mal (Myelin and Lymphocyte Protein) and two isoforms of Mobp (Myelin-Associated Oligodendrocyte Basic Protein), in addition to Mbp (Myelin Basic Protein), which is shown for comparison. (B) Among neuronal factors, this included Hapln1, Hapln2, Hapln3, Hapln4 (Hyaluronan and Proteoglycan Link proteins), Prnp (Prion protein) and Rtn4 (NoGo-A). Only the Prion protein mRNA was dysregulated in the opposite direction in Atxn2 -KO tissue. (C) Among factors of unclear origin and without membrane association, this included Rgs8 (Regulator of G Protein Signaling 8) and Klk6 (Kallikrein Related Peptidase 6). Analysis was performed for KO mice at the age of 6 months when obesity is evident, comparing with CAG100Hom mice at the age of 14 months when weight loss is strong. Fold-changes are shown within the bars, t-test was used in general. (D) Schematic representation of the dysregulated transcripts and proteins in the cerebellum of 14-month-old CAG100Hom mouse.

Article Snippet: The TaqMan ® Assays utilized for this study are: Atxn2 (Mm01199894_m1), Calb1 (Mm00486647-m1), Cnp (Mm01306641_m1), Hapln1 (Mm00618325_m1), Hapln2 (Mm00480745_m1), Hapln3 (Mm00724203_m1), Hapln4 (Mm00625974_m1), Hprt1 (Mm00446968_m1), Klk6 (Mm00478322_m1), Mag (Mm00487538_m1), Mal (Mm01339780_m1), Mbp (Mm01266402_m1), Mobp long transcripts (Mm02745649_m1), Mobp short transcripts (Mm01348317_g1), Mog (Mm00447824_m 1), Nefh (Mm01191456_m1), Nefl (Mm01315666_m1), Nefm (Mm00456201_m1), Nptn (Mm00485990_m1), Plp1 (Mm01297210_m1), Prnp (Mm004483 89_m 1), Rgs8 (Mm01290239_m1), Rtn4 (Mm00445861_m1), Snca (Mm00447333_m1), Tbp (Mm00446973_m1), Tuba4a (Mm00849767_s1).

Techniques: