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  • 93
    BioVendor Instruments gfap
    a) Illustration shows the location of the hippocampus at Bregma +1.77mm. b) Bar chart shows the absorbance of <t>GFAP,</t> measured using an <t>ELISA</t> kit. In the hippocampus Bonferroni analysis revealed higher GFAP levels in the WT females compared with WT males.
    Gfap, supplied by BioVendor Instruments, used in various techniques. Bioz Stars score: 93/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore glial fibrillar acid protein gfap
    a) Illustration shows the location of the hippocampus at Bregma +1.77mm. b) Bar chart shows the absorbance of <t>GFAP,</t> measured using an <t>ELISA</t> kit. In the hippocampus Bonferroni analysis revealed higher GFAP levels in the WT females compared with WT males.
    Glial Fibrillar Acid Protein Gfap, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies glial fibrillar acid protein gfap
    Full thickness brain biopsy of case 2. The H E stain (A), immunostaining for glial fibrillar acid protein (B), axons (neurofilament cocktail) (C) and myelin (Luxol fast blue/cresyl violet) (D) shows a mild pallor of the myelin and reduction of axon density towards the deep white matter (separated by a yellow dotted line in D) where frequent axonal spheroids are seen (inset in E). The axonal spheroids label with antibodies for <t>neurofilaments</t> (E), amyloid precursor protein (F) and ubiquitin (G). Increased numbers of CD68 positive microglial cells are present in the deeper white matter (H), which show yellow-light brown cytoplasm on H E and negative control sections and appear blue when viewed as a negative colour inversion image (insets in H). Scale bar: 1 mm in A–D, 5 µm in E–H, 10 µm insets in E and H.
    Glial Fibrillar Acid Protein Gfap, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 811 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore mouse anti glial fibrillar acid protein
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker BLBP (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker <t>SOX-2</t> (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Mouse Anti Glial Fibrillar Acid Protein, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Quanterix gfap
    <t>GFAP</t> <t>CSF</t> and serum levels in multiple sclerosis patients and patients with other non-inflammatory neurological diseases (OND). GFAP: glial fibrillary acidic protein. CSF: cerebrospinal fluid, PMS: progressive multiple sclerosis, RRMS: relapsing-remitting multiple sclerosis. P-values were calculated with Kruskal-Wallis test followed by Dunn’s multiple comparison tests. * p
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    93
    Neuromics gfap
    PDE11A4 is selectively expressed in neurons. Sagittal sections (~2.64 mm lateral from Bregma) were taken from <t>PDE11A</t> wild-type (WT) and knockout (KO) mice and stained for PDE11A4 (PD11A-112; green), nuclei (DAPI; blue), and a marker for neurons (neuron specific enolase, NSE; red), astrocytes (glial fibrillary acidic protein, <t>GFAP;</t> red), dendrites (MAP2; red), or axon bundles (myelin basic protein, MBP; red). (a) Staining for PDE11A4 is far stronger in hippocampi of PDE11A WT mice vs KO mice, arguing for specificity of the antibody. Further, PDE11A4 expression was far stronger in the ventral hippocampal formation (VHIPP) of PDE11A WT mice vs the dorsal hippocampal formation (DHIPP) of PDE11A WT mice, consistent with previous reports using in situ hybridization for mRNA and western blotting for protein. PDE11A4 protein can be seen in the cell body layer and throughout stratum radiatum of CA1. At the anterior edge of the CA1 field (facing left), a slightly more intense patch of staining that extends throughout stratum radiatum in the shape of a narrow triangle is reliably observed across animals. The anatomical localization of this staining suggests it may actually reflect CA2, although it is thought that CA2 is minimally present in VHIPP. Labeling in the cell body layer and stratum radiatum of ventral CA1 abruptly stops anteriorly at the border for CA3 and dorsally at the stratum lacunosum of dentate gyrus. Labeling for PDE11A4 can also be seen in the axon bundle projecting out of the hippocampus. (b) A closer view shows that PDE11A4 is expressed in a subset of neuronal cell bodies, particularly those neurons lying adjacent to the stratum radiatum. (c) In contrast, PDE11A4 does not appear to be expressed in astrocytes. (d) Consistent with its expression throughout the stratum radiatum, PDE11A4 protein expression colocalizes in some instances with the dendritic marker MAP2. (e) PDE11A4 protein expression also colocalizes with MBP. PDE11A4 protein expression in axons is consistent with the fact that faint PDE11A4 protein expression can be measured when western blotting brain regions that, themselves, do not express PDE11A mRNA but do receive projections from the hippocampus (eg, prefrontal cortex and striatum). Histogram stretch and gamma adjustments applied uniformly across PDE11A KO and WT sections for graphical clarity of images.
    Gfap, supplied by Neuromics, used in various techniques. Bioz Stars score: 93/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    OriGene gfap
    Optimization of transient expression for WT and AxD-associated <t>GFAP</t> mutant proteins in SW13vim- cells. ( A ) Western blot of SW13vim- cells transfected for 24 hr with the designated GFAP constructs. NTC, non-transfected control. Top and bottom blots show GFAP and pan-actin, respectively, in the Triton X-100-soluble fraction (TX-100). Middle blot is a total cell lysate (TCL) blot of GFAP from the same <t>transfections.</t> ( B ) Corresponding immunofluorescence staining of GFAP in SW13vim- cells after 24 hr of transfection. Scale bars = 10 µm.
    Gfap, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Biorbyt gfap
    Optimization of transient expression for WT and AxD-associated <t>GFAP</t> mutant proteins in SW13vim- cells. ( A ) Western blot of SW13vim- cells transfected for 24 hr with the designated GFAP constructs. NTC, non-transfected control. Top and bottom blots show GFAP and pan-actin, respectively, in the Triton X-100-soluble fraction (TX-100). Middle blot is a total cell lysate (TCL) blot of GFAP from the same <t>transfections.</t> ( B ) Corresponding immunofluorescence staining of GFAP in SW13vim- cells after 24 hr of transfection. Scale bars = 10 µm.
    Gfap, supplied by Biorbyt, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    The Jackson Laboratory gfap promoter gfap cre
    Effects of global ERK1 and neuronal ERK2 deletion on pERK immunoreactivity at 15 min post‐30 min HI insult A – C , control (ERK1/2 WT ) animal ( A ), global deletion of ERK1 and ERK2 WT (ERK1 KO ) ( B ), global ERK1 deletion and homozygous neuronal ERK2 deletion (ERK1 KO ERK2 ΔSyn ) ( C ). C , pERK immunoreactivity is almost completely reduced following deletion of both copies of ERK1 and ERK2. D–I , quantification and distribution of pERK immunoreactivity at high magnification. D , pERK staining in the contralateral pyriform cortex of ERK1/2 WT with strong neuronal reactivity and prominent dendritic staining which disappears in the presence of global ERK1 deletion and homozygous neuronal ERK2 mutation ERK1 KO ERK2 ΔSyn . E – I , residual immunoreactivity on the ipsilateral side. E and H , pERK alone. F and I , immunofluorescence double labelling with <t>GFAP</t> demonstrating co‐localisation of pERK in astrocytes, particularly pronounced in ERK1 WT ERK2 ΔSyn (white arrows). Scale bar = 25 µm [Color figure can be viewed at <t>http://wileyonlinelibrary.com</t> ]
    Gfap Promoter Gfap Cre, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    gfap  (Abcam)
    94
    Abcam gfap
    Approximate onset and duration of reactive astrogliosis, impairment of perivascular <t>AQP4</t> polarity and accumulation of p-tau in different brain regions following TBI. The horizontal bars intend to illustrate the approximate onset and extent of the changes in <t>GFAP</t> and p-tau immunoreactivities, and impairment of AQP4 polarity. The increase in shading intensity reflects an increased severity of these pathological changes.
    Gfap, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 3032 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Synaptic Systems gfap 134b1
    Approximate onset and duration of reactive astrogliosis, impairment of perivascular <t>AQP4</t> polarity and accumulation of p-tau in different brain regions following TBI. The horizontal bars intend to illustrate the approximate onset and extent of the changes in <t>GFAP</t> and p-tau immunoreactivities, and impairment of AQP4 polarity. The increase in shading intensity reflects an increased severity of these pathological changes.
    Gfap 134b1, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam gfap ab233621
    Approximate onset and duration of reactive astrogliosis, impairment of perivascular <t>AQP4</t> polarity and accumulation of p-tau in different brain regions following TBI. The horizontal bars intend to illustrate the approximate onset and extent of the changes in <t>GFAP</t> and p-tau immunoreactivities, and impairment of AQP4 polarity. The increase in shading intensity reflects an increased severity of these pathological changes.
    Gfap Ab233621, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore gfap cy3
    ENPs labeled by Sox10 -H2BVenus expression exhibit stem cell characteristics Sox10 -H2BVenus+ cells from 14dpc fetal gut give rise to neurospheres in culture ( a ) and express the neural crest stem cell markers p75 LNTR ( b ) and nestin ( c ) detected by <t>Cy3</t> (red) IHC labeling (200X). Sox10 -H2BVenus+ cells isolated from 14dpc gut grown in low density cultures give rise to multipotent colonies ( d ) that contain neurons (peripherin+), glia <t>(GFAP+)</t> and myofibroblasts (SMA+) labeled by triple immunofluorescence, 100X magnification. Detection of myofibroblasts with SMA-FITC resulted in co-visualization of SMA+ cells with H2BVenus+ nuclei in multipotent (SMA panel) colonies. RT-PCR detects expression ( e ) of stem cell genes (Abcg2, Bmi1, Dll1, Klf4, Lgr5, Msi1, Myc, Nes, Ngfr, Notch1, Pou5f1, Sox2), neural crest genes ( FoxD3, Snai1, Snai2, Sox9, Sox10, Twist1 ), neuronal progenitor marker genes ( Ascl1, Neurod1, Neurog1, Phox2b, Prph1, Uchl1 ), and peripheral glia markers ( Erbb3, Fabp7, Gfap, Mpz, Nrg1, Nrtn, Olig2, S100b ) in Sox10 -H2BVenus+ enteric populations at discrete developmental stages. Housekeeping control genes ( Actb, Ipo8, Ubc ) are shown for comparison. Lanes include 100bp Molecular Weight Marker (M), no template control (H 2 O), total 14.5dpc fetal mouse RNA (Total E14), flow-sorted 14.5dpc gut H2BVenus+ ENPs (E14 Sox10+), cultured neurospheres from 14.5dpc H2BVenus+ gut (E14 NS), postnatal day 6 H2BVenus+ cells isolated from gut muscle strips (P6 GMS).
    Gfap Cy3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies gfap mo761
    ENPs labeled by Sox10 -H2BVenus expression exhibit stem cell characteristics Sox10 -H2BVenus+ cells from 14dpc fetal gut give rise to neurospheres in culture ( a ) and express the neural crest stem cell markers p75 LNTR ( b ) and nestin ( c ) detected by <t>Cy3</t> (red) IHC labeling (200X). Sox10 -H2BVenus+ cells isolated from 14dpc gut grown in low density cultures give rise to multipotent colonies ( d ) that contain neurons (peripherin+), glia <t>(GFAP+)</t> and myofibroblasts (SMA+) labeled by triple immunofluorescence, 100X magnification. Detection of myofibroblasts with SMA-FITC resulted in co-visualization of SMA+ cells with H2BVenus+ nuclei in multipotent (SMA panel) colonies. RT-PCR detects expression ( e ) of stem cell genes (Abcg2, Bmi1, Dll1, Klf4, Lgr5, Msi1, Myc, Nes, Ngfr, Notch1, Pou5f1, Sox2), neural crest genes ( FoxD3, Snai1, Snai2, Sox9, Sox10, Twist1 ), neuronal progenitor marker genes ( Ascl1, Neurod1, Neurog1, Phox2b, Prph1, Uchl1 ), and peripheral glia markers ( Erbb3, Fabp7, Gfap, Mpz, Nrg1, Nrtn, Olig2, S100b ) in Sox10 -H2BVenus+ enteric populations at discrete developmental stages. Housekeeping control genes ( Actb, Ipo8, Ubc ) are shown for comparison. Lanes include 100bp Molecular Weight Marker (M), no template control (H 2 O), total 14.5dpc fetal mouse RNA (Total E14), flow-sorted 14.5dpc gut H2BVenus+ ENPs (E14 Sox10+), cultured neurospheres from 14.5dpc H2BVenus+ gut (E14 NS), postnatal day 6 H2BVenus+ cells isolated from gut muscle strips (P6 GMS).
    Gfap Mo761, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology gfap pe
    Decreased expression of <t>DCX</t> in the infected brain. Cells isolated from MCMV-infected and control brains were analyzed for intracellular doublecortin (DCX) and glial fibrillary acidic protein <t>(GFAP)</t> at 7 d p.i. Histogram overlays from isotype (grey line, filled), infected (blue line, filled), and control (red line, tinge) are shown for: A . DCX, a marker for young/immature neurons and B . GFAP, a marker for glial precursors. Infected brains showed reduced expression levels of intracellular DCX compared to control brains while there was no difference in the expression levels of GFAP in MCMV infected versus control brains. C . Data were derived from 3 independent experiments, n = 3–5 neonates. * p
    Gfap Pe, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Agilent technologies poly gfap
    Decreased expression of <t>DCX</t> in the infected brain. Cells isolated from MCMV-infected and control brains were analyzed for intracellular doublecortin (DCX) and glial fibrillary acidic protein <t>(GFAP)</t> at 7 d p.i. Histogram overlays from isotype (grey line, filled), infected (blue line, filled), and control (red line, tinge) are shown for: A . DCX, a marker for young/immature neurons and B . GFAP, a marker for glial precursors. Infected brains showed reduced expression levels of intracellular DCX compared to control brains while there was no difference in the expression levels of GFAP in MCMV infected versus control brains. C . Data were derived from 3 independent experiments, n = 3–5 neonates. * p
    Poly Gfap, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher murine gfap
    Decreased expression of <t>DCX</t> in the infected brain. Cells isolated from MCMV-infected and control brains were analyzed for intracellular doublecortin (DCX) and glial fibrillary acidic protein <t>(GFAP)</t> at 7 d p.i. Histogram overlays from isotype (grey line, filled), infected (blue line, filled), and control (red line, tinge) are shown for: A . DCX, a marker for young/immature neurons and B . GFAP, a marker for glial precursors. Infected brains showed reduced expression levels of intracellular DCX compared to control brains while there was no difference in the expression levels of GFAP in MCMV infected versus control brains. C . Data were derived from 3 independent experiments, n = 3–5 neonates. * p
    Murine Gfap, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Agilent technologies unconjugated gfap
    Transient expression of NFIA in neuroepithelial stem cells confers glial competency. A , Quantitative PCR of candidate genes associated with glial competency treated in serum (1% FBS) conditions for 30 days. **One-way ANOVA (p-value = 0.025, n= 3 biologically independent experiments, mean values are represented by a black bar). B , Overexpression of NFIA leads to profound morphological changes within 5 days of doxycycline treatment marked by yellow arrowheads (n= 5 biologically independent experiments). C , Quantitative PCR analysis of <t>GFAP</t> and NFIA expression in NSCs treated with doxycycline for 5 days and subsequent removal for an additional 3 and 5 days or continuous treatment (+dox) (n= 3 biologically independent experiments, mean values are represented in bar graph). D , Intracellular FACS analysis for GFAP and <t>CD44</t> during the differentiation of NFIA-induced NSCs at 56 (p6) and 77 (p8). E , Quantification of the percentage of GFAP expressing cells at different timepoints (n=3 biologically independent experiments, mean values are represented in bar graph). F , Immunofluorescence staining of GFAP and SLC1A2 in d60 astrocyte culture (n= 5 biologically independent experiments). G , Quantitative PCR analysis of genes associated with NSCs, neurons, astrocytes and oligodendrocytes from NFIA-induced astrocytes. H , Heatmap of normalized read-counts representing genes associated with astrocyte identity ( Supplemental Table 1 ). Yellow = Zhang et al., purple = TCW et al., brown = Santos et al., green = this study. Scale bars are 50 μ m. Error bars are calculated by S.E.M.
    Unconjugated Gfap, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cell Signaling Technology Inc gfap
    Effects of brain cells in FUS-MB treated 5XFAD a Representative images of neuronal loss in the entorhinal cortex by H E and TUNEL staining. Neuronal damage was observed in the entorhinal cortex of dCLN ligated 5XFAD, but no neuronal damage was observed in dCLN ligated 5XFAD after FUS-MB. b . Representative merged images of <t>GFAP</t> (green), <t>Aβ</t> (red), and DAPI (blue) immunostaining in hippocampus and entorhinal cortex (left). The isolated images of GFAP to compare the changes of reactive astrocytes (right). Reactive astrocytes were reduced only in entorhinal cortex after FUS-MB. c . Representative merged images of Iba1(red), Aβ (green), and DAPI (blue) immunostaining (left). The isolated images of Iba1 to compare the changes of microglia activation (right). Microglial activation was reduced in the entire brain by FUS-MB.
    Gfap, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1799 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    a) Illustration shows the location of the hippocampus at Bregma +1.77mm. b) Bar chart shows the absorbance of GFAP, measured using an ELISA kit. In the hippocampus Bonferroni analysis revealed higher GFAP levels in the WT females compared with WT males.

    Journal: Brain research

    Article Title: Sex effects of Interleukin-6 deficiency on neuroinflammation in aged C57Bl/6 mice

    doi: 10.1016/j.brainres.2009.12.091

    Figure Lengend Snippet: a) Illustration shows the location of the hippocampus at Bregma +1.77mm. b) Bar chart shows the absorbance of GFAP, measured using an ELISA kit. In the hippocampus Bonferroni analysis revealed higher GFAP levels in the WT females compared with WT males.

    Article Snippet: A commercially available sandwich ELISA was used to detect GFAP (Biovendor).

    Techniques: Enzyme-linked Immunosorbent Assay

    Full thickness brain biopsy of case 2. The H E stain (A), immunostaining for glial fibrillar acid protein (B), axons (neurofilament cocktail) (C) and myelin (Luxol fast blue/cresyl violet) (D) shows a mild pallor of the myelin and reduction of axon density towards the deep white matter (separated by a yellow dotted line in D) where frequent axonal spheroids are seen (inset in E). The axonal spheroids label with antibodies for neurofilaments (E), amyloid precursor protein (F) and ubiquitin (G). Increased numbers of CD68 positive microglial cells are present in the deeper white matter (H), which show yellow-light brown cytoplasm on H E and negative control sections and appear blue when viewed as a negative colour inversion image (insets in H). Scale bar: 1 mm in A–D, 5 µm in E–H, 10 µm insets in E and H.

    Journal: Journal of Neurology, Neurosurgery, and Psychiatry

    Article Title: Hereditary leukoencephalopathy with axonal spheroids: a spectrum of phenotypes from CNS vasculitis to parkinsonism in an adult onset leukodystrophy series

    doi: 10.1136/jnnp-2015-310788

    Figure Lengend Snippet: Full thickness brain biopsy of case 2. The H E stain (A), immunostaining for glial fibrillar acid protein (B), axons (neurofilament cocktail) (C) and myelin (Luxol fast blue/cresyl violet) (D) shows a mild pallor of the myelin and reduction of axon density towards the deep white matter (separated by a yellow dotted line in D) where frequent axonal spheroids are seen (inset in E). The axonal spheroids label with antibodies for neurofilaments (E), amyloid precursor protein (F) and ubiquitin (G). Increased numbers of CD68 positive microglial cells are present in the deeper white matter (H), which show yellow-light brown cytoplasm on H E and negative control sections and appear blue when viewed as a negative colour inversion image (insets in H). Scale bar: 1 mm in A–D, 5 µm in E–H, 10 µm insets in E and H.

    Article Snippet: Sections were examined by immunohistochemistry with the following antibodies: glial fibrillar acid protein (GFAP) (polyclonal, 1:2500, Dako), phosphorylated neurofilaments (clone SMI31, 1:5000, Sternberg), neurofilament cocktail (clone 2F11, 1:500, Dako/Cappel), myelin basic protein (clone SMI94, 1:2000, Sternberger), amyloid precursor protein (clone 22C11, 1:800, Chemicon/Millipore), amyloid-β (clone 6F3D, 1:100, Dako), ubiquitin (polyclonal, 1:1200, Dako), p62 (3/P62LCK Ligand, 1:100, BD Transduction), α-synuclein (clone KM51, 1:50, Leica/Novocastra), hyperphosphorylated τ (clone AT8, 1:1200, INNOGENETICS), TDP-43 (clone 2E2-D3, 1:3000, Abnova), CD68 (clone PG-M1, 1:100, Dako), CD3 (LN10, 1:100, Leica/Novocastra), CD20 (clone 7D1, 1:200, Dako).

    Techniques: Staining, Immunostaining, Negative Control

    Full thickness brain biopsy of case 1. The H E stained section (A) shows frequent axonal swellings in the subcortical white matter (inset). Immunostaining for glial fibrillar acid protein (B) reveals a severe chronic fibrillar and reactive stellate astrogliosis in the subcortical white matter and in the cortex. Immunostaining for myelin basic protein with SMI94 antibody (C) and of axons with SMI31 antibody (D) reveals no apparent myelin or axon loss. SMI31 immunoreactive axonal spheroids are frequent (E) while periodic acid-Schiff (PAS) positive pigmented glial cells (F, red arrowhead) are sparse. Axonal spheroids are positive for p62 (G), amyloid precursor protein (H) and amyloid-β (I), and negative for α-synuclein (J). Occasional scattered PAS-positive cells in the white matter (K, red arrowhead) show positive labelling for the macrophage lysosome marker CD68 (L). A negative control section (M) highlights the yellow-brown pigment in the cytoplasm of these monocyte-derived cells (brown arrowheads), which appears blue in a negative—complete colour inversion image (N, blue arrowheads). Scale bar: 1 mm in A–D, 10 µm inset in A, 50 µm in E–F, 50 µm in G–N.

    Journal: Journal of Neurology, Neurosurgery, and Psychiatry

    Article Title: Hereditary leukoencephalopathy with axonal spheroids: a spectrum of phenotypes from CNS vasculitis to parkinsonism in an adult onset leukodystrophy series

    doi: 10.1136/jnnp-2015-310788

    Figure Lengend Snippet: Full thickness brain biopsy of case 1. The H E stained section (A) shows frequent axonal swellings in the subcortical white matter (inset). Immunostaining for glial fibrillar acid protein (B) reveals a severe chronic fibrillar and reactive stellate astrogliosis in the subcortical white matter and in the cortex. Immunostaining for myelin basic protein with SMI94 antibody (C) and of axons with SMI31 antibody (D) reveals no apparent myelin or axon loss. SMI31 immunoreactive axonal spheroids are frequent (E) while periodic acid-Schiff (PAS) positive pigmented glial cells (F, red arrowhead) are sparse. Axonal spheroids are positive for p62 (G), amyloid precursor protein (H) and amyloid-β (I), and negative for α-synuclein (J). Occasional scattered PAS-positive cells in the white matter (K, red arrowhead) show positive labelling for the macrophage lysosome marker CD68 (L). A negative control section (M) highlights the yellow-brown pigment in the cytoplasm of these monocyte-derived cells (brown arrowheads), which appears blue in a negative—complete colour inversion image (N, blue arrowheads). Scale bar: 1 mm in A–D, 10 µm inset in A, 50 µm in E–F, 50 µm in G–N.

    Article Snippet: Sections were examined by immunohistochemistry with the following antibodies: glial fibrillar acid protein (GFAP) (polyclonal, 1:2500, Dako), phosphorylated neurofilaments (clone SMI31, 1:5000, Sternberg), neurofilament cocktail (clone 2F11, 1:500, Dako/Cappel), myelin basic protein (clone SMI94, 1:2000, Sternberger), amyloid precursor protein (clone 22C11, 1:800, Chemicon/Millipore), amyloid-β (clone 6F3D, 1:100, Dako), ubiquitin (polyclonal, 1:1200, Dako), p62 (3/P62LCK Ligand, 1:100, BD Transduction), α-synuclein (clone KM51, 1:50, Leica/Novocastra), hyperphosphorylated τ (clone AT8, 1:1200, INNOGENETICS), TDP-43 (clone 2E2-D3, 1:3000, Abnova), CD68 (clone PG-M1, 1:100, Dako), CD3 (LN10, 1:100, Leica/Novocastra), CD20 (clone 7D1, 1:200, Dako).

    Techniques: Staining, Immunostaining, Marker, Negative Control, Derivative Assay

    WT and PAR1 -/- mice show similar vasculature characteristics. ( A and B ) Factor VIII immunostained and hematoxylin counterstained WT ( A, n = 5 animals) and PAR1 -/- ( B, n = 5 animals) sections from striatum. Arrows indicate examples of Factor VIII immunostained vascular profiles. ( C and D ) GFAP immunostained and hematoxylin counterstained WT ( C, n = 5 animals) and PAR1 -/- ( D, n = 5 animals) cortical brain sections. ( Insets ) Electron microscopic images of a portion of a WT ( C ) and PAR1 -/- ( D ) capillary showing GFAP immunoreactivity around capillary. ( E and F ) Electron microscopic image of WT ( E, n = 5 animals) and PAR1 -/- ( F, n = 5 animals) brain capillary from striatum. ( G ) Quantification of vascular density (capillaries per mm 2 ) and capillary lumen area (μm 2 ) in WT ( n = 5) and PAR1 -/- ( n = 5) mice ( P > 0.05, unpaired t tests). Error measurements are SEM. ( H ) Average infarct volume was significantly reduced after intracerebroventricular injection of 6 mM PAR1 antagonist BMS-200261 ( n = 6, P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: The contribution of protease-activated receptor 1 to neuronal damage caused by transient focal cerebral ischemia

    doi: 10.1073/pnas.2235594100

    Figure Lengend Snippet: WT and PAR1 -/- mice show similar vasculature characteristics. ( A and B ) Factor VIII immunostained and hematoxylin counterstained WT ( A, n = 5 animals) and PAR1 -/- ( B, n = 5 animals) sections from striatum. Arrows indicate examples of Factor VIII immunostained vascular profiles. ( C and D ) GFAP immunostained and hematoxylin counterstained WT ( C, n = 5 animals) and PAR1 -/- ( D, n = 5 animals) cortical brain sections. ( Insets ) Electron microscopic images of a portion of a WT ( C ) and PAR1 -/- ( D ) capillary showing GFAP immunoreactivity around capillary. ( E and F ) Electron microscopic image of WT ( E, n = 5 animals) and PAR1 -/- ( F, n = 5 animals) brain capillary from striatum. ( G ) Quantification of vascular density (capillaries per mm 2 ) and capillary lumen area (μm 2 ) in WT ( n = 5) and PAR1 -/- ( n = 5) mice ( P > 0.05, unpaired t tests). Error measurements are SEM. ( H ) Average infarct volume was significantly reduced after intracerebroventricular injection of 6 mM PAR1 antagonist BMS-200261 ( n = 6, P

    Article Snippet: Sections were incubated at room temperature with Factor VIII antibody (prediluted, rabbit polyclonal, DAKO) or GFAP (1:100, mouse monoclonal, DAKO) and appropriate blocking reagents (animal research kit, DAKO).

    Techniques: Mouse Assay, Injection

    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker BLBP (A’, red). Merged images (A ″ ). Immuno-staining for GFAP (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).

    Journal: Hippocampus

    Article Title: Identification of a Sustained Neurogenic Zone at the Dorsal Surface of the Adult Mouse Hippocampus and Its Regulation by the Chemokine SDF-1

    doi: 10.1002/hipo.22428

    Figure Lengend Snippet: CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker BLBP (A’, red). Merged images (A ″ ). Immuno-staining for GFAP (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).

    Article Snippet: Briefly, sections were blocked in phosphate buffer containing 0.1% Triton X-100 and incubated overnight at 4 °C with the following primary antibodies: CD45 (1/300, rat, Millipore, CA) and Iba-1 (1/300, rabbit, Wako Chemicals USA, VA) for microglia; CD45 and F4/80 (1/300, rabbit, Santa Cruz Biotechnology) for macrophages; Nestin (1/150, rat, BD Pharmingen, CA) for early neural progenitors, SOX-2 (1/200, rabbit, Millipore, CA) for neuronal stem cells, GFAP (1/300, mouse, Sigma-Aldrich, MO) and BLBP (1/100, rabbit, Millipore) for radial glia, DCX [1:700; Guina pig, Millipore, CA) for migratory neuroblasts, NeuN (1/300, mouse, Milli-pore, MA), Prox-1 (1/500, rabbit, Millipore, CA)] for DG granular neurons, calretinin and calbindin (1/250, rabbit, Millipore, MA) for mature DG neurons, laminin (1/100, rabbit, Millipore, CA), vWF (1/100, rabbit, Santa Cruz Biotechnology, CA) for blood vessels.

    Techniques: Staining, Marker, Immunostaining, Expressing

    Expression of EGFP in a previously unrecognized region with characteristics of NSCs in the adult brain of CXCR4-EGFP tg mice. A-A ″ , a-a ″ : Experimental protocol used to dissect the dorsal surface of the hippocampus (DSH) from the brain in adult mice. The hippocampus was dissected as shown in (A-A ″ ), and the DSH was displayed (a). (a’, a ″ ) DSH viewed at low power under an fluorescent binocular stereomicroscope equipped with a black and white CCD camera showing bright GFP fluorescence (a’), as shown by green dots in (a ″ ). B-B ″ : Confocal reconstruction of representative whole mount preparations from areas highlighted in (a’, a ″ ), showing strong EGFP fluorescence in the SHZ. B: A face view of the DSH showing the location of the SHZ, also (B’). B ″ : Horizontal sections through the DSH showing strong EGFP expression (green) in the SHZ (left side of image) and the DG (right side of image) with some CXCR4-EGFP cells scattered between the SHZ toward the DG (B ″ ), exhibiting long uni-or dual processes, suggestive of their capacity to migrate in direction to the DG (B ″ ). Boxed areas in panels (C, C’, and C ″ ) show representative higher magnification views of CXCR4-EGFP cells at the transitional area between the SHZ and the DG. C: Shows densely compacted cells in the SHZ with no apparent extracellular space. C’ and C ″ : Shows streams of CXCR4- EGFP cells sometimes with long processes. D, D’, and D ″ : Representative higher magnification image of these cells costained with the Tbr-2 antibody, indicating that these cells are neuronal progenitors. E–I: Whole mount preparations of the SHZ stained for EGFP antibody (green) showing CXCR4-EGFP cells also expressing a wide variety of stem cell markers (red). E–E ″ : Example of a whole mount SHZ preparation stained for the stem/progenitor marker nestin (red), showing that virtually all the EGFP cells (E) costained for nestin (E’); Merged image (E ″ ). Whole mount SHZ preparations (F–F ″ , G–G ″ ) and cross sections (F ‴ , G ‴ ) showing that all SHZ cells (F, G), costained for GFAP (red, F) and the neuronal stem marker SOX-2 (red, G’); (F ″ , F ‴ , and G ″ , G ‴ ) are merged images. To detect proliferating cells, animals were injected with BrdU, two times a day for 1 week, and euthanized 24 h later. Whole mount SHZ preparation (H-H ″ ) and cross sections (H ‴ ) show that virtually all EGFP cells (H, I’, green) incorporated BrdU (H’, H ″ , H ‴ , I, red), and also coexpressed the cell cycle marker Ki67 (I’, I ″ ). K: Neurospheres (NSs) obtained from cultures of SHZ whole mount preparations, indicating their self-renewal potential in cultures. Immunostaining of SHZ NSs for CXCR4 (K’, K ‴ ) and nestin (K ″ , K ‴ ) demonstrates uniform expression of CXCR4 (blue) and nestin (red), further indicating the identity of SHZ cells as progenitor cells in cultures. L: Chemokines such as SDF-1 increased [Ca 2+ ] i in whole NSs, indicating the expression of functional CXCR4 receptors. (L’) Example of three cells dissociated from SHZ NSs that show responses to SDF-1. Addition of another chemokine such MCP-1 also increased [Ca 2+ ] i as did the purinergic receptor agonist ATP. Scale bars are 2 mm in panels (A-A ″ ), 500 μm in panels a-a ″ , 250 μm in panels (B-B ″ ), 100 μm in panels (EE ″ , F-F ″ , G-G ‴ , H-H ‴ ), 100 μm in panels (I-I ″ ), and 20 μm in panels (C-C ‴ ), and (D-D ‴ ’, F ‴ , G ‴ , and H ‴ .]

    Journal: Hippocampus

    Article Title: Identification of a Sustained Neurogenic Zone at the Dorsal Surface of the Adult Mouse Hippocampus and Its Regulation by the Chemokine SDF-1

    doi: 10.1002/hipo.22428

    Figure Lengend Snippet: Expression of EGFP in a previously unrecognized region with characteristics of NSCs in the adult brain of CXCR4-EGFP tg mice. A-A ″ , a-a ″ : Experimental protocol used to dissect the dorsal surface of the hippocampus (DSH) from the brain in adult mice. The hippocampus was dissected as shown in (A-A ″ ), and the DSH was displayed (a). (a’, a ″ ) DSH viewed at low power under an fluorescent binocular stereomicroscope equipped with a black and white CCD camera showing bright GFP fluorescence (a’), as shown by green dots in (a ″ ). B-B ″ : Confocal reconstruction of representative whole mount preparations from areas highlighted in (a’, a ″ ), showing strong EGFP fluorescence in the SHZ. B: A face view of the DSH showing the location of the SHZ, also (B’). B ″ : Horizontal sections through the DSH showing strong EGFP expression (green) in the SHZ (left side of image) and the DG (right side of image) with some CXCR4-EGFP cells scattered between the SHZ toward the DG (B ″ ), exhibiting long uni-or dual processes, suggestive of their capacity to migrate in direction to the DG (B ″ ). Boxed areas in panels (C, C’, and C ″ ) show representative higher magnification views of CXCR4-EGFP cells at the transitional area between the SHZ and the DG. C: Shows densely compacted cells in the SHZ with no apparent extracellular space. C’ and C ″ : Shows streams of CXCR4- EGFP cells sometimes with long processes. D, D’, and D ″ : Representative higher magnification image of these cells costained with the Tbr-2 antibody, indicating that these cells are neuronal progenitors. E–I: Whole mount preparations of the SHZ stained for EGFP antibody (green) showing CXCR4-EGFP cells also expressing a wide variety of stem cell markers (red). E–E ″ : Example of a whole mount SHZ preparation stained for the stem/progenitor marker nestin (red), showing that virtually all the EGFP cells (E) costained for nestin (E’); Merged image (E ″ ). Whole mount SHZ preparations (F–F ″ , G–G ″ ) and cross sections (F ‴ , G ‴ ) showing that all SHZ cells (F, G), costained for GFAP (red, F) and the neuronal stem marker SOX-2 (red, G’); (F ″ , F ‴ , and G ″ , G ‴ ) are merged images. To detect proliferating cells, animals were injected with BrdU, two times a day for 1 week, and euthanized 24 h later. Whole mount SHZ preparation (H-H ″ ) and cross sections (H ‴ ) show that virtually all EGFP cells (H, I’, green) incorporated BrdU (H’, H ″ , H ‴ , I, red), and also coexpressed the cell cycle marker Ki67 (I’, I ″ ). K: Neurospheres (NSs) obtained from cultures of SHZ whole mount preparations, indicating their self-renewal potential in cultures. Immunostaining of SHZ NSs for CXCR4 (K’, K ‴ ) and nestin (K ″ , K ‴ ) demonstrates uniform expression of CXCR4 (blue) and nestin (red), further indicating the identity of SHZ cells as progenitor cells in cultures. L: Chemokines such as SDF-1 increased [Ca 2+ ] i in whole NSs, indicating the expression of functional CXCR4 receptors. (L’) Example of three cells dissociated from SHZ NSs that show responses to SDF-1. Addition of another chemokine such MCP-1 also increased [Ca 2+ ] i as did the purinergic receptor agonist ATP. Scale bars are 2 mm in panels (A-A ″ ), 500 μm in panels a-a ″ , 250 μm in panels (B-B ″ ), 100 μm in panels (EE ″ , F-F ″ , G-G ‴ , H-H ‴ ), 100 μm in panels (I-I ″ ), and 20 μm in panels (C-C ‴ ), and (D-D ‴ ’, F ‴ , G ‴ , and H ‴ .]

    Article Snippet: Briefly, sections were blocked in phosphate buffer containing 0.1% Triton X-100 and incubated overnight at 4 °C with the following primary antibodies: CD45 (1/300, rat, Millipore, CA) and Iba-1 (1/300, rabbit, Wako Chemicals USA, VA) for microglia; CD45 and F4/80 (1/300, rabbit, Santa Cruz Biotechnology) for macrophages; Nestin (1/150, rat, BD Pharmingen, CA) for early neural progenitors, SOX-2 (1/200, rabbit, Millipore, CA) for neuronal stem cells, GFAP (1/300, mouse, Sigma-Aldrich, MO) and BLBP (1/100, rabbit, Millipore) for radial glia, DCX [1:700; Guina pig, Millipore, CA) for migratory neuroblasts, NeuN (1/300, mouse, Milli-pore, MA), Prox-1 (1/500, rabbit, Millipore, CA)] for DG granular neurons, calretinin and calbindin (1/250, rabbit, Millipore, MA) for mature DG neurons, laminin (1/100, rabbit, Millipore, CA), vWF (1/100, rabbit, Santa Cruz Biotechnology, CA) for blood vessels.

    Techniques: Expressing, Mouse Assay, Fluorescence, Staining, Marker, Injection, Immunostaining, Functional Assay

    GFAP CSF and serum levels in multiple sclerosis patients and patients with other non-inflammatory neurological diseases (OND). GFAP: glial fibrillary acidic protein. CSF: cerebrospinal fluid, PMS: progressive multiple sclerosis, RRMS: relapsing-remitting multiple sclerosis. P-values were calculated with Kruskal-Wallis test followed by Dunn’s multiple comparison tests. * p

    Journal: Scientific Reports

    Article Title: Serum GFAP as a biomarker for disease severity in multiple sclerosis

    doi: 10.1038/s41598-018-33158-8

    Figure Lengend Snippet: GFAP CSF and serum levels in multiple sclerosis patients and patients with other non-inflammatory neurological diseases (OND). GFAP: glial fibrillary acidic protein. CSF: cerebrospinal fluid, PMS: progressive multiple sclerosis, RRMS: relapsing-remitting multiple sclerosis. P-values were calculated with Kruskal-Wallis test followed by Dunn’s multiple comparison tests. * p

    Article Snippet: GFAP and NfL measurements Both, GFAP and NfL were measured in CSF and serum using the Simoa technology and GFAP Discovery and NfL Early Access assays (Quanterix Corporation, Lexington, MA, USA).

    Techniques:

    Receiver Operating Characteristic Curve Results for Biomarkers at the Acute Postinjury Time Point and 24 to 48 Hours After Injury AUC indicates area under the curve; GFAP, glial fibrillary acidic protein; NF-L, neurofilament light chain; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Journal: JAMA Network Open

    Article Title: Association of Blood Biomarkers With Acute Sport-Related Concussion in Collegiate Athletes

    doi: 10.1001/jamanetworkopen.2019.19771

    Figure Lengend Snippet: Receiver Operating Characteristic Curve Results for Biomarkers at the Acute Postinjury Time Point and 24 to 48 Hours After Injury AUC indicates area under the curve; GFAP, glial fibrillary acidic protein; NF-L, neurofilament light chain; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Article Snippet: Glial fibrillary acidic protein (GFAP), ubiquitin C-terminal hydrolase-L1 (UCH-L1), neurofilament light chain, and tau were quantified using the Quanterix Simoa multiplex assay.

    Techniques:

    Receiver Operating Characteristic Curve Results for Biomarkers Differentiating Athletes With Concussion With Either Loss of Consciousness (LOC) or Posttraumatic Amnesia (PTA) (LOC-PTA) From Contact Sport Controls and Non–Contact Sport Controls at the Acute Postinjury Time Point and 24 to 48 Hours After Injury AUC indicates area under the curve; GFAP, glial fibrillary acidic protein; NF-L, neurofilament light chain; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Journal: JAMA Network Open

    Article Title: Association of Blood Biomarkers With Acute Sport-Related Concussion in Collegiate Athletes

    doi: 10.1001/jamanetworkopen.2019.19771

    Figure Lengend Snippet: Receiver Operating Characteristic Curve Results for Biomarkers Differentiating Athletes With Concussion With Either Loss of Consciousness (LOC) or Posttraumatic Amnesia (PTA) (LOC-PTA) From Contact Sport Controls and Non–Contact Sport Controls at the Acute Postinjury Time Point and 24 to 48 Hours After Injury AUC indicates area under the curve; GFAP, glial fibrillary acidic protein; NF-L, neurofilament light chain; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Article Snippet: Glial fibrillary acidic protein (GFAP), ubiquitin C-terminal hydrolase-L1 (UCH-L1), neurofilament light chain, and tau were quantified using the Quanterix Simoa multiplex assay.

    Techniques:

    Baseline and Postinjury Biomarker Levels in Athletes With Concussion With Either Loss of Consciousness (LOC) or Posttraumatic Amnesia (PTA) (LOC-PTA), Athletes With Concussion With No LOC or PTA (No LOC-PTA), Contact Sport Controls, and Non–Contact Sport Controls Biomarker levels represent natural log (ln) transformed scale. Error bars indicate SEs. GFAP indicates glial fibrillary acidic protein; NF-L, neurofilament light chain; RTP, return to play; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Journal: JAMA Network Open

    Article Title: Association of Blood Biomarkers With Acute Sport-Related Concussion in Collegiate Athletes

    doi: 10.1001/jamanetworkopen.2019.19771

    Figure Lengend Snippet: Baseline and Postinjury Biomarker Levels in Athletes With Concussion With Either Loss of Consciousness (LOC) or Posttraumatic Amnesia (PTA) (LOC-PTA), Athletes With Concussion With No LOC or PTA (No LOC-PTA), Contact Sport Controls, and Non–Contact Sport Controls Biomarker levels represent natural log (ln) transformed scale. Error bars indicate SEs. GFAP indicates glial fibrillary acidic protein; NF-L, neurofilament light chain; RTP, return to play; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Article Snippet: Glial fibrillary acidic protein (GFAP), ubiquitin C-terminal hydrolase-L1 (UCH-L1), neurofilament light chain, and tau were quantified using the Quanterix Simoa multiplex assay.

    Techniques: Biomarker Assay, Transformation Assay

    Baseline and Postinjury Biomarker Levels in the Concussion, Contact Sport Control, and Non–Contact Sport Control Groups Biomarker levels represent natural log (ln) transformed scale. Error bars indicate SEs. GFAP indicates glial fibrillary acidic protein; NF-L, neurofilament light chain; RTP, return to play; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Journal: JAMA Network Open

    Article Title: Association of Blood Biomarkers With Acute Sport-Related Concussion in Collegiate Athletes

    doi: 10.1001/jamanetworkopen.2019.19771

    Figure Lengend Snippet: Baseline and Postinjury Biomarker Levels in the Concussion, Contact Sport Control, and Non–Contact Sport Control Groups Biomarker levels represent natural log (ln) transformed scale. Error bars indicate SEs. GFAP indicates glial fibrillary acidic protein; NF-L, neurofilament light chain; RTP, return to play; and UCH-L1, ubiquitin C-terminal hydrolase-L1.

    Article Snippet: Glial fibrillary acidic protein (GFAP), ubiquitin C-terminal hydrolase-L1 (UCH-L1), neurofilament light chain, and tau were quantified using the Quanterix Simoa multiplex assay.

    Techniques: Biomarker Assay, Transformation Assay

    PDE11A4 is selectively expressed in neurons. Sagittal sections (~2.64 mm lateral from Bregma) were taken from PDE11A wild-type (WT) and knockout (KO) mice and stained for PDE11A4 (PD11A-112; green), nuclei (DAPI; blue), and a marker for neurons (neuron specific enolase, NSE; red), astrocytes (glial fibrillary acidic protein, GFAP; red), dendrites (MAP2; red), or axon bundles (myelin basic protein, MBP; red). (a) Staining for PDE11A4 is far stronger in hippocampi of PDE11A WT mice vs KO mice, arguing for specificity of the antibody. Further, PDE11A4 expression was far stronger in the ventral hippocampal formation (VHIPP) of PDE11A WT mice vs the dorsal hippocampal formation (DHIPP) of PDE11A WT mice, consistent with previous reports using in situ hybridization for mRNA and western blotting for protein. PDE11A4 protein can be seen in the cell body layer and throughout stratum radiatum of CA1. At the anterior edge of the CA1 field (facing left), a slightly more intense patch of staining that extends throughout stratum radiatum in the shape of a narrow triangle is reliably observed across animals. The anatomical localization of this staining suggests it may actually reflect CA2, although it is thought that CA2 is minimally present in VHIPP. Labeling in the cell body layer and stratum radiatum of ventral CA1 abruptly stops anteriorly at the border for CA3 and dorsally at the stratum lacunosum of dentate gyrus. Labeling for PDE11A4 can also be seen in the axon bundle projecting out of the hippocampus. (b) A closer view shows that PDE11A4 is expressed in a subset of neuronal cell bodies, particularly those neurons lying adjacent to the stratum radiatum. (c) In contrast, PDE11A4 does not appear to be expressed in astrocytes. (d) Consistent with its expression throughout the stratum radiatum, PDE11A4 protein expression colocalizes in some instances with the dendritic marker MAP2. (e) PDE11A4 protein expression also colocalizes with MBP. PDE11A4 protein expression in axons is consistent with the fact that faint PDE11A4 protein expression can be measured when western blotting brain regions that, themselves, do not express PDE11A mRNA but do receive projections from the hippocampus (eg, prefrontal cortex and striatum). Histogram stretch and gamma adjustments applied uniformly across PDE11A KO and WT sections for graphical clarity of images.

    Journal: Neuropsychopharmacology

    Article Title: Phosphodiesterase 11A (PDE11A), Enriched in Ventral Hippocampus Neurons, is Required for Consolidation of Social but not Nonsocial Memories in Mice

    doi: 10.1038/npp.2016.106

    Figure Lengend Snippet: PDE11A4 is selectively expressed in neurons. Sagittal sections (~2.64 mm lateral from Bregma) were taken from PDE11A wild-type (WT) and knockout (KO) mice and stained for PDE11A4 (PD11A-112; green), nuclei (DAPI; blue), and a marker for neurons (neuron specific enolase, NSE; red), astrocytes (glial fibrillary acidic protein, GFAP; red), dendrites (MAP2; red), or axon bundles (myelin basic protein, MBP; red). (a) Staining for PDE11A4 is far stronger in hippocampi of PDE11A WT mice vs KO mice, arguing for specificity of the antibody. Further, PDE11A4 expression was far stronger in the ventral hippocampal formation (VHIPP) of PDE11A WT mice vs the dorsal hippocampal formation (DHIPP) of PDE11A WT mice, consistent with previous reports using in situ hybridization for mRNA and western blotting for protein. PDE11A4 protein can be seen in the cell body layer and throughout stratum radiatum of CA1. At the anterior edge of the CA1 field (facing left), a slightly more intense patch of staining that extends throughout stratum radiatum in the shape of a narrow triangle is reliably observed across animals. The anatomical localization of this staining suggests it may actually reflect CA2, although it is thought that CA2 is minimally present in VHIPP. Labeling in the cell body layer and stratum radiatum of ventral CA1 abruptly stops anteriorly at the border for CA3 and dorsally at the stratum lacunosum of dentate gyrus. Labeling for PDE11A4 can also be seen in the axon bundle projecting out of the hippocampus. (b) A closer view shows that PDE11A4 is expressed in a subset of neuronal cell bodies, particularly those neurons lying adjacent to the stratum radiatum. (c) In contrast, PDE11A4 does not appear to be expressed in astrocytes. (d) Consistent with its expression throughout the stratum radiatum, PDE11A4 protein expression colocalizes in some instances with the dendritic marker MAP2. (e) PDE11A4 protein expression also colocalizes with MBP. PDE11A4 protein expression in axons is consistent with the fact that faint PDE11A4 protein expression can be measured when western blotting brain regions that, themselves, do not express PDE11A mRNA but do receive projections from the hippocampus (eg, prefrontal cortex and striatum). Histogram stretch and gamma adjustments applied uniformly across PDE11A KO and WT sections for graphical clarity of images.

    Article Snippet: PBT) 3 × 10 min and then either PBT/10% BlokHen (Aves BH-1001—Tigard, OR) or PBT/5% milk for 1 h. Slides were probed overnight at 4 °C with antibodies against PDE11A (1:100 of FabGennix PD11–112- Frisco, TX), PDE11A4 (1:10 000 of Aves PDE11A#1), NSE (1:500 of Aves), MBP (1:100 of Aves), GFAP (1:500 of Aves), or MAP2 (1:2000 of Neuromics- CH22103—Minneapolis, MN).

    Techniques: Knock-Out, Mouse Assay, Staining, Marker, Expressing, In Situ Hybridization, Western Blot, Labeling

    Optimization of transient expression for WT and AxD-associated GFAP mutant proteins in SW13vim- cells. ( A ) Western blot of SW13vim- cells transfected for 24 hr with the designated GFAP constructs. NTC, non-transfected control. Top and bottom blots show GFAP and pan-actin, respectively, in the Triton X-100-soluble fraction (TX-100). Middle blot is a total cell lysate (TCL) blot of GFAP from the same transfections. ( B ) Corresponding immunofluorescence staining of GFAP in SW13vim- cells after 24 hr of transfection. Scale bars = 10 µm.

    Journal: eLife

    Article Title: Site-specific phosphorylation and caspase cleavage of GFAP are new markers of Alexander disease severity

    doi: 10.7554/eLife.47789

    Figure Lengend Snippet: Optimization of transient expression for WT and AxD-associated GFAP mutant proteins in SW13vim- cells. ( A ) Western blot of SW13vim- cells transfected for 24 hr with the designated GFAP constructs. NTC, non-transfected control. Top and bottom blots show GFAP and pan-actin, respectively, in the Triton X-100-soluble fraction (TX-100). Middle blot is a total cell lysate (TCL) blot of GFAP from the same transfections. ( B ) Corresponding immunofluorescence staining of GFAP in SW13vim- cells after 24 hr of transfection. Scale bars = 10 µm.

    Article Snippet: Site directed mutagenesis, in vitro assembly, transfections, and immunofluorescence Mutagenesis of GFAP (Origene, Rockville, MD, in vector CMV6-XL6) was performed using the QuikChange II mutagenesis kit (Agilent) to generate the designated point mutants.

    Techniques: Expressing, Mutagenesis, Western Blot, Transfection, Construct, Immunofluorescence, Staining

    Effects of global ERK1 and neuronal ERK2 deletion on pERK immunoreactivity at 15 min post‐30 min HI insult A – C , control (ERK1/2 WT ) animal ( A ), global deletion of ERK1 and ERK2 WT (ERK1 KO ) ( B ), global ERK1 deletion and homozygous neuronal ERK2 deletion (ERK1 KO ERK2 ΔSyn ) ( C ). C , pERK immunoreactivity is almost completely reduced following deletion of both copies of ERK1 and ERK2. D–I , quantification and distribution of pERK immunoreactivity at high magnification. D , pERK staining in the contralateral pyriform cortex of ERK1/2 WT with strong neuronal reactivity and prominent dendritic staining which disappears in the presence of global ERK1 deletion and homozygous neuronal ERK2 mutation ERK1 KO ERK2 ΔSyn . E – I , residual immunoreactivity on the ipsilateral side. E and H , pERK alone. F and I , immunofluorescence double labelling with GFAP demonstrating co‐localisation of pERK in astrocytes, particularly pronounced in ERK1 WT ERK2 ΔSyn (white arrows). Scale bar = 25 µm [Color figure can be viewed at http://wileyonlinelibrary.com ]

    Journal: The Journal of Physiology

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury

    doi: 10.1113/JP275649

    Figure Lengend Snippet: Effects of global ERK1 and neuronal ERK2 deletion on pERK immunoreactivity at 15 min post‐30 min HI insult A – C , control (ERK1/2 WT ) animal ( A ), global deletion of ERK1 and ERK2 WT (ERK1 KO ) ( B ), global ERK1 deletion and homozygous neuronal ERK2 deletion (ERK1 KO ERK2 ΔSyn ) ( C ). C , pERK immunoreactivity is almost completely reduced following deletion of both copies of ERK1 and ERK2. D–I , quantification and distribution of pERK immunoreactivity at high magnification. D , pERK staining in the contralateral pyriform cortex of ERK1/2 WT with strong neuronal reactivity and prominent dendritic staining which disappears in the presence of global ERK1 deletion and homozygous neuronal ERK2 mutation ERK1 KO ERK2 ΔSyn . E – I , residual immunoreactivity on the ipsilateral side. E and H , pERK alone. F and I , immunofluorescence double labelling with GFAP demonstrating co‐localisation of pERK in astrocytes, particularly pronounced in ERK1 WT ERK2 ΔSyn (white arrows). Scale bar = 25 µm [Color figure can be viewed at http://wileyonlinelibrary.com ]

    Article Snippet: Syn‐Cre mice were provided by Dr Axel Behrens from the Mammalian Genetics Laboratory, Cancer Research UK, and animals expressing Cre recombinase under the control of GFAP promoter (GFAP‐Cre) were from Jackson Labs (USA, http://jaxmice.jax.org/strain/004600.html ).

    Techniques: Staining, Mutagenesis, Immunofluorescence

    IGFR knockout in astrocytes decreases glucose and Aβ uptake. (A) Astroyctes were treated with 1 mM of 2D-glucose (2DG) and accumulation of 2DG6P was measured using luminescence. IGFR-KO astrocytes were significantly impaired in glucose uptake compared to GFP and un-transduced (UT) controls. (B) Flow cytometry for Aβ 1−42 -555 uptake showed reduced total fluorescence intensity in IGFR-KO (CRE) culture compared to GFP control culture. Unstained astrocytes were used for forward and side scatter gating. (C) Geometric mean of the fluorescence intensity of 555 in GFP + cells was also significantly reduced in IGFR-KO astrocytes compared to controls. (D) Representative confocal images of astrocytes (n = 4/group) treated with Aβ 1−42 -555 show reduced uptake of Aβ 1−42 in IGFR-KO astrocytes (Lower panel) compared to GFP controls (Upper panel). GFP fluorescence represents endogenous expression of viral transduction while staining for GFAP was performed by immunocytochemistry. Scale bar = 20 μm; n = 5/group Data (A–C) are represented as the mean ± SEM; *P

    Journal: Molecular Metabolism

    Article Title: Insulin-like growth factor receptor signaling regulates working memory, mitochondrial metabolism, and amyloid-β uptake in astrocytes

    doi: 10.1016/j.molmet.2018.01.013

    Figure Lengend Snippet: IGFR knockout in astrocytes decreases glucose and Aβ uptake. (A) Astroyctes were treated with 1 mM of 2D-glucose (2DG) and accumulation of 2DG6P was measured using luminescence. IGFR-KO astrocytes were significantly impaired in glucose uptake compared to GFP and un-transduced (UT) controls. (B) Flow cytometry for Aβ 1−42 -555 uptake showed reduced total fluorescence intensity in IGFR-KO (CRE) culture compared to GFP control culture. Unstained astrocytes were used for forward and side scatter gating. (C) Geometric mean of the fluorescence intensity of 555 in GFP + cells was also significantly reduced in IGFR-KO astrocytes compared to controls. (D) Representative confocal images of astrocytes (n = 4/group) treated with Aβ 1−42 -555 show reduced uptake of Aβ 1−42 in IGFR-KO astrocytes (Lower panel) compared to GFP controls (Upper panel). GFP fluorescence represents endogenous expression of viral transduction while staining for GFAP was performed by immunocytochemistry. Scale bar = 20 μm; n = 5/group Data (A–C) are represented as the mean ± SEM; *P

    Article Snippet: Young (4–6 months) and old (22–24 months) C57Bl/6 mice were obtained from the NIA colony at the National Institutes of Health and Igfr f/f (B6;129-Igf1rtm2Arge/J; loxP sites flanking exon 3) and GFAP-Cre (B6.Cg-TgGFAP-cre/ERT2/505Fmv/J) mice were obtained from Jackson laboratories.

    Techniques: Knock-Out, Flow Cytometry, Cytometry, Fluorescence, Expressing, Transduction, Staining, Immunocytochemistry

    IGFR signaling deficiency modulates mitochondrial energy charge in astrocytes in complete growth media. Knockout of IGF-1R in astrocytes (A–C) showed 30% reduction in mRNA levels (A), ∼20% reduction in energy charge (B) with no significant change in mitochondrial DNA/nuclear DNA ratio (C) in the overall population. (D) Representative western blots for IGFRβ and ERK showing reduction IGFR in the knockout and reduced p44 ERK levels quantified in (E), while no differences were found in AKT phosphorylation. Data were analyzed by two-tailed student T-test; Mean ± SEM; n = 5 per group. (F) Representative confocal images of untransduced (UT; top panel), GFP-transduced (GFP; middle panel) and IGF-1R knockout (CRE; bottom panel) astrocytes labeled for GFAP (blue) and Mitotracker (Red) showing a vesicular perinuclear mitochondrial localization in the knockout. Scale bar = 20 μm; n = 3/group.

    Journal: Molecular Metabolism

    Article Title: Insulin-like growth factor receptor signaling regulates working memory, mitochondrial metabolism, and amyloid-β uptake in astrocytes

    doi: 10.1016/j.molmet.2018.01.013

    Figure Lengend Snippet: IGFR signaling deficiency modulates mitochondrial energy charge in astrocytes in complete growth media. Knockout of IGF-1R in astrocytes (A–C) showed 30% reduction in mRNA levels (A), ∼20% reduction in energy charge (B) with no significant change in mitochondrial DNA/nuclear DNA ratio (C) in the overall population. (D) Representative western blots for IGFRβ and ERK showing reduction IGFR in the knockout and reduced p44 ERK levels quantified in (E), while no differences were found in AKT phosphorylation. Data were analyzed by two-tailed student T-test; Mean ± SEM; n = 5 per group. (F) Representative confocal images of untransduced (UT; top panel), GFP-transduced (GFP; middle panel) and IGF-1R knockout (CRE; bottom panel) astrocytes labeled for GFAP (blue) and Mitotracker (Red) showing a vesicular perinuclear mitochondrial localization in the knockout. Scale bar = 20 μm; n = 3/group.

    Article Snippet: Young (4–6 months) and old (22–24 months) C57Bl/6 mice were obtained from the NIA colony at the National Institutes of Health and Igfr f/f (B6;129-Igf1rtm2Arge/J; loxP sites flanking exon 3) and GFAP-Cre (B6.Cg-TgGFAP-cre/ERT2/505Fmv/J) mice were obtained from Jackson laboratories.

    Techniques: Knock-Out, Western Blot, Two Tailed Test, Labeling

    Age-dependent impairment in learning is associated with decreased hippocampal IGFR expression and gliosis. C57Bl/6 young (4–6 m) and old (22–24 m) mice were tested on the radial arm water maze for spatial learning (n = 10/group). Old mice showed significantly more errors (A) and pathlength (B) to target and display a shallow learning curve compared to young mice. Hippocampal IGFR (C) expression was significantly decreased, while gliosis marker, GFAP, was significantly increased (D) in old mice compared to young. Increase in GFAP protein expression is validated via western blotting (E; n = 6/group) as well as by immunohistochemistry (n = 3/group) in young and old mice. Scale bar = 100 μm. Data are represented as the Mean ± SEM. *P

    Journal: Molecular Metabolism

    Article Title: Insulin-like growth factor receptor signaling regulates working memory, mitochondrial metabolism, and amyloid-β uptake in astrocytes

    doi: 10.1016/j.molmet.2018.01.013

    Figure Lengend Snippet: Age-dependent impairment in learning is associated with decreased hippocampal IGFR expression and gliosis. C57Bl/6 young (4–6 m) and old (22–24 m) mice were tested on the radial arm water maze for spatial learning (n = 10/group). Old mice showed significantly more errors (A) and pathlength (B) to target and display a shallow learning curve compared to young mice. Hippocampal IGFR (C) expression was significantly decreased, while gliosis marker, GFAP, was significantly increased (D) in old mice compared to young. Increase in GFAP protein expression is validated via western blotting (E; n = 6/group) as well as by immunohistochemistry (n = 3/group) in young and old mice. Scale bar = 100 μm. Data are represented as the Mean ± SEM. *P

    Article Snippet: Young (4–6 months) and old (22–24 months) C57Bl/6 mice were obtained from the NIA colony at the National Institutes of Health and Igfr f/f (B6;129-Igf1rtm2Arge/J; loxP sites flanking exon 3) and GFAP-Cre (B6.Cg-TgGFAP-cre/ERT2/505Fmv/J) mice were obtained from Jackson laboratories.

    Techniques: Expressing, Mouse Assay, Marker, Western Blot, Immunohistochemistry

    Approximate onset and duration of reactive astrogliosis, impairment of perivascular AQP4 polarity and accumulation of p-tau in different brain regions following TBI. The horizontal bars intend to illustrate the approximate onset and extent of the changes in GFAP and p-tau immunoreactivities, and impairment of AQP4 polarity. The increase in shading intensity reflects an increased severity of these pathological changes.

    Journal: Scientific Reports

    Article Title: Perivascular AQP4 dysregulation in the hippocampal CA1 area after traumatic brain injury is alleviated by adenosine A2A receptor inactivation

    doi: 10.1038/s41598-017-02505-6

    Figure Lengend Snippet: Approximate onset and duration of reactive astrogliosis, impairment of perivascular AQP4 polarity and accumulation of p-tau in different brain regions following TBI. The horizontal bars intend to illustrate the approximate onset and extent of the changes in GFAP and p-tau immunoreactivities, and impairment of AQP4 polarity. The increase in shading intensity reflects an increased severity of these pathological changes.

    Article Snippet: Primary antibody incubations were conducted overnight for A2A R (Abcam; 1:200), AQP4 (Abcam; 1:200), GFAP (Abcam,1:200), total tau (Tau5), p-tau (Thr205) (Pierce; 1:100), p-tau (Ser262) (Pierce; 1:100), p-tau (Ser404) (Abcam; 1:100), and AT8 (Pierce; 1:100), or T22 (Abcam; 1:100).

    Techniques:

    Loss of AQP4 polarity after brain injury in a mouse model of TBI and patients’ brains. ( a ) In the mouse TBI model, compared with the SHAM group, reactive astrogliosis (asterisk) was still obvious 4 weeks after the injury. AQP4 immunoreactivity was confined to the perivascular end feet (arrows) in the contralateral hippocampal CA1 region of the SHAM WT mice ( b ) and the SHAM A 2A R KO mice ( c ). A large proportion of AQP4 shifted to the soma and coarse processes of the GFAP-positive reactive astrocytes 7 d ( d ) and 4 weeks ( e ) after TBI. AQP4 dysregulation was alleviated by A 2A R KO ( f,g ). The arrows indicate the relative normal AQP4 distribution, and the arrowheads indicate AQP4 dysregulation. Scale bar: 50 μm. ( h ) Quantification of AQP4 polarity in the SHAM group and 7 d and 4 weeks post-TBI group. n = 4 per group. Data represent mean ± s.e.m., * p

    Journal: Scientific Reports

    Article Title: Perivascular AQP4 dysregulation in the hippocampal CA1 area after traumatic brain injury is alleviated by adenosine A2A receptor inactivation

    doi: 10.1038/s41598-017-02505-6

    Figure Lengend Snippet: Loss of AQP4 polarity after brain injury in a mouse model of TBI and patients’ brains. ( a ) In the mouse TBI model, compared with the SHAM group, reactive astrogliosis (asterisk) was still obvious 4 weeks after the injury. AQP4 immunoreactivity was confined to the perivascular end feet (arrows) in the contralateral hippocampal CA1 region of the SHAM WT mice ( b ) and the SHAM A 2A R KO mice ( c ). A large proportion of AQP4 shifted to the soma and coarse processes of the GFAP-positive reactive astrocytes 7 d ( d ) and 4 weeks ( e ) after TBI. AQP4 dysregulation was alleviated by A 2A R KO ( f,g ). The arrows indicate the relative normal AQP4 distribution, and the arrowheads indicate AQP4 dysregulation. Scale bar: 50 μm. ( h ) Quantification of AQP4 polarity in the SHAM group and 7 d and 4 weeks post-TBI group. n = 4 per group. Data represent mean ± s.e.m., * p

    Article Snippet: Primary antibody incubations were conducted overnight for A2A R (Abcam; 1:200), AQP4 (Abcam; 1:200), GFAP (Abcam,1:200), total tau (Tau5), p-tau (Thr205) (Pierce; 1:100), p-tau (Ser262) (Pierce; 1:100), p-tau (Ser404) (Abcam; 1:100), and AT8 (Pierce; 1:100), or T22 (Abcam; 1:100).

    Techniques: Mouse Assay

    ENPs labeled by Sox10 -H2BVenus expression exhibit stem cell characteristics Sox10 -H2BVenus+ cells from 14dpc fetal gut give rise to neurospheres in culture ( a ) and express the neural crest stem cell markers p75 LNTR ( b ) and nestin ( c ) detected by Cy3 (red) IHC labeling (200X). Sox10 -H2BVenus+ cells isolated from 14dpc gut grown in low density cultures give rise to multipotent colonies ( d ) that contain neurons (peripherin+), glia (GFAP+) and myofibroblasts (SMA+) labeled by triple immunofluorescence, 100X magnification. Detection of myofibroblasts with SMA-FITC resulted in co-visualization of SMA+ cells with H2BVenus+ nuclei in multipotent (SMA panel) colonies. RT-PCR detects expression ( e ) of stem cell genes (Abcg2, Bmi1, Dll1, Klf4, Lgr5, Msi1, Myc, Nes, Ngfr, Notch1, Pou5f1, Sox2), neural crest genes ( FoxD3, Snai1, Snai2, Sox9, Sox10, Twist1 ), neuronal progenitor marker genes ( Ascl1, Neurod1, Neurog1, Phox2b, Prph1, Uchl1 ), and peripheral glia markers ( Erbb3, Fabp7, Gfap, Mpz, Nrg1, Nrtn, Olig2, S100b ) in Sox10 -H2BVenus+ enteric populations at discrete developmental stages. Housekeeping control genes ( Actb, Ipo8, Ubc ) are shown for comparison. Lanes include 100bp Molecular Weight Marker (M), no template control (H 2 O), total 14.5dpc fetal mouse RNA (Total E14), flow-sorted 14.5dpc gut H2BVenus+ ENPs (E14 Sox10+), cultured neurospheres from 14.5dpc H2BVenus+ gut (E14 NS), postnatal day 6 H2BVenus+ cells isolated from gut muscle strips (P6 GMS).

    Journal: Genesis (New York, N.Y. : 2000)

    Article Title: Isolation and Live Imaging of Enteric Progenitors based on Sox10-Histone2BVenus Transgene Expression

    doi: 10.1002/dvg.20748

    Figure Lengend Snippet: ENPs labeled by Sox10 -H2BVenus expression exhibit stem cell characteristics Sox10 -H2BVenus+ cells from 14dpc fetal gut give rise to neurospheres in culture ( a ) and express the neural crest stem cell markers p75 LNTR ( b ) and nestin ( c ) detected by Cy3 (red) IHC labeling (200X). Sox10 -H2BVenus+ cells isolated from 14dpc gut grown in low density cultures give rise to multipotent colonies ( d ) that contain neurons (peripherin+), glia (GFAP+) and myofibroblasts (SMA+) labeled by triple immunofluorescence, 100X magnification. Detection of myofibroblasts with SMA-FITC resulted in co-visualization of SMA+ cells with H2BVenus+ nuclei in multipotent (SMA panel) colonies. RT-PCR detects expression ( e ) of stem cell genes (Abcg2, Bmi1, Dll1, Klf4, Lgr5, Msi1, Myc, Nes, Ngfr, Notch1, Pou5f1, Sox2), neural crest genes ( FoxD3, Snai1, Snai2, Sox9, Sox10, Twist1 ), neuronal progenitor marker genes ( Ascl1, Neurod1, Neurog1, Phox2b, Prph1, Uchl1 ), and peripheral glia markers ( Erbb3, Fabp7, Gfap, Mpz, Nrg1, Nrtn, Olig2, S100b ) in Sox10 -H2BVenus+ enteric populations at discrete developmental stages. Housekeeping control genes ( Actb, Ipo8, Ubc ) are shown for comparison. Lanes include 100bp Molecular Weight Marker (M), no template control (H 2 O), total 14.5dpc fetal mouse RNA (Total E14), flow-sorted 14.5dpc gut H2BVenus+ ENPs (E14 Sox10+), cultured neurospheres from 14.5dpc H2BVenus+ gut (E14 NS), postnatal day 6 H2BVenus+ cells isolated from gut muscle strips (P6 GMS).

    Article Snippet: Cultures were washed, fixed, and stained with antibodies to Peripherin (Chemicon, 1:1,000), GFAP-Cy3 (Sigma, 1:800) and αSMA-FITC (Sigma, 1:800).

    Techniques: Labeling, Expressing, Immunohistochemistry, Isolation, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction, Marker, Molecular Weight, Flow Cytometry, Cell Culture

    Decreased expression of DCX in the infected brain. Cells isolated from MCMV-infected and control brains were analyzed for intracellular doublecortin (DCX) and glial fibrillary acidic protein (GFAP) at 7 d p.i. Histogram overlays from isotype (grey line, filled), infected (blue line, filled), and control (red line, tinge) are shown for: A . DCX, a marker for young/immature neurons and B . GFAP, a marker for glial precursors. Infected brains showed reduced expression levels of intracellular DCX compared to control brains while there was no difference in the expression levels of GFAP in MCMV infected versus control brains. C . Data were derived from 3 independent experiments, n = 3–5 neonates. * p

    Journal: PLoS ONE

    Article Title: Murine Cytomegalovirus Infection of Neural Stem Cells Alters Neurogenesis in the Developing Brain

    doi: 10.1371/journal.pone.0016211

    Figure Lengend Snippet: Decreased expression of DCX in the infected brain. Cells isolated from MCMV-infected and control brains were analyzed for intracellular doublecortin (DCX) and glial fibrillary acidic protein (GFAP) at 7 d p.i. Histogram overlays from isotype (grey line, filled), infected (blue line, filled), and control (red line, tinge) are shown for: A . DCX, a marker for young/immature neurons and B . GFAP, a marker for glial precursors. Infected brains showed reduced expression levels of intracellular DCX compared to control brains while there was no difference in the expression levels of GFAP in MCMV infected versus control brains. C . Data were derived from 3 independent experiments, n = 3–5 neonates. * p

    Article Snippet: Surface markers were selected based on the experiment being performed: nestin-PE or APC (R & D Systems, Minneapolis, MN and BD Biosciences, San Jose, CA), DCX, GFAP-PE (Santa Cruz Biotechnology, Santa Cruz, CA) and Oct4-APC (R & D Systems, Minneapolis, MN).

    Techniques: Expressing, Infection, Isolation, Marker, Derivative Assay

    Transient expression of NFIA in neuroepithelial stem cells confers glial competency. A , Quantitative PCR of candidate genes associated with glial competency treated in serum (1% FBS) conditions for 30 days. **One-way ANOVA (p-value = 0.025, n= 3 biologically independent experiments, mean values are represented by a black bar). B , Overexpression of NFIA leads to profound morphological changes within 5 days of doxycycline treatment marked by yellow arrowheads (n= 5 biologically independent experiments). C , Quantitative PCR analysis of GFAP and NFIA expression in NSCs treated with doxycycline for 5 days and subsequent removal for an additional 3 and 5 days or continuous treatment (+dox) (n= 3 biologically independent experiments, mean values are represented in bar graph). D , Intracellular FACS analysis for GFAP and CD44 during the differentiation of NFIA-induced NSCs at 56 (p6) and 77 (p8). E , Quantification of the percentage of GFAP expressing cells at different timepoints (n=3 biologically independent experiments, mean values are represented in bar graph). F , Immunofluorescence staining of GFAP and SLC1A2 in d60 astrocyte culture (n= 5 biologically independent experiments). G , Quantitative PCR analysis of genes associated with NSCs, neurons, astrocytes and oligodendrocytes from NFIA-induced astrocytes. H , Heatmap of normalized read-counts representing genes associated with astrocyte identity ( Supplemental Table 1 ). Yellow = Zhang et al., purple = TCW et al., brown = Santos et al., green = this study. Scale bars are 50 μ m. Error bars are calculated by S.E.M.

    Journal: Nature biotechnology

    Article Title: NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells

    doi: 10.1038/s41587-019-0035-0

    Figure Lengend Snippet: Transient expression of NFIA in neuroepithelial stem cells confers glial competency. A , Quantitative PCR of candidate genes associated with glial competency treated in serum (1% FBS) conditions for 30 days. **One-way ANOVA (p-value = 0.025, n= 3 biologically independent experiments, mean values are represented by a black bar). B , Overexpression of NFIA leads to profound morphological changes within 5 days of doxycycline treatment marked by yellow arrowheads (n= 5 biologically independent experiments). C , Quantitative PCR analysis of GFAP and NFIA expression in NSCs treated with doxycycline for 5 days and subsequent removal for an additional 3 and 5 days or continuous treatment (+dox) (n= 3 biologically independent experiments, mean values are represented in bar graph). D , Intracellular FACS analysis for GFAP and CD44 during the differentiation of NFIA-induced NSCs at 56 (p6) and 77 (p8). E , Quantification of the percentage of GFAP expressing cells at different timepoints (n=3 biologically independent experiments, mean values are represented in bar graph). F , Immunofluorescence staining of GFAP and SLC1A2 in d60 astrocyte culture (n= 5 biologically independent experiments). G , Quantitative PCR analysis of genes associated with NSCs, neurons, astrocytes and oligodendrocytes from NFIA-induced astrocytes. H , Heatmap of normalized read-counts representing genes associated with astrocyte identity ( Supplemental Table 1 ). Yellow = Zhang et al., purple = TCW et al., brown = Santos et al., green = this study. Scale bars are 50 μ m. Error bars are calculated by S.E.M.

    Article Snippet: Alexa 647 conjugated CD44 (Biolegend) and unconjugated GFAP (Dako) was added to the cells as described by the manufacturer and incubated for 40 minutes on ice.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Significance Assay, Over Expression, FACS, Immunofluorescence, Staining

    NFIA cannot maintain the glial competent state. A , FACS plot of CD44 expressing cells treated with continuous doxycycline or doxycycline removed demonstrates that CD44 expression is lost after doxycycline removal. B , Immunofluorescence staining for NFIA, GFAP and TUBB3 in NSCs, NFIA-induced NSCs and NFIA-induced NSCs with doxycycline removal (n= 3 biologically independent experiments). C , Schematic representation of cells induced with NFIA and attaining glial competency then reversal to glial incompetency with doxycycline withdrawal. D , Quantitative PCR data of NFIA expression along the time course represented in C. E , Sample distance plot for RNA expression of NSCs at different timepoints related C. F , Sample distance plot for chromatin accessibility compared to glial competent NSCs (gcNSCs) and hPSC-derived astrocytes (200 days of in vitro culture). G , Example ATAC-seq tracks at the GFAP locus that depicts the lack of chromatin accessibility in several NSC samples. H , Bisulfite sequencing of the promoter region of the GFAP promoter (P1 and P2) suggests that CD44 positive cells resulting from overexpression of NFIA leads to demethylation of a specific CpG on the GFAP promoter (circled in green).

    Journal: Nature biotechnology

    Article Title: NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells

    doi: 10.1038/s41587-019-0035-0

    Figure Lengend Snippet: NFIA cannot maintain the glial competent state. A , FACS plot of CD44 expressing cells treated with continuous doxycycline or doxycycline removed demonstrates that CD44 expression is lost after doxycycline removal. B , Immunofluorescence staining for NFIA, GFAP and TUBB3 in NSCs, NFIA-induced NSCs and NFIA-induced NSCs with doxycycline removal (n= 3 biologically independent experiments). C , Schematic representation of cells induced with NFIA and attaining glial competency then reversal to glial incompetency with doxycycline withdrawal. D , Quantitative PCR data of NFIA expression along the time course represented in C. E , Sample distance plot for RNA expression of NSCs at different timepoints related C. F , Sample distance plot for chromatin accessibility compared to glial competent NSCs (gcNSCs) and hPSC-derived astrocytes (200 days of in vitro culture). G , Example ATAC-seq tracks at the GFAP locus that depicts the lack of chromatin accessibility in several NSC samples. H , Bisulfite sequencing of the promoter region of the GFAP promoter (P1 and P2) suggests that CD44 positive cells resulting from overexpression of NFIA leads to demethylation of a specific CpG on the GFAP promoter (circled in green).

    Article Snippet: Alexa 647 conjugated CD44 (Biolegend) and unconjugated GFAP (Dako) was added to the cells as described by the manufacturer and incubated for 40 minutes on ice.

    Techniques: FACS, Expressing, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, RNA Expression, Derivative Assay, In Vitro, Methylation Sequencing, Over Expression

    Effects of brain cells in FUS-MB treated 5XFAD a Representative images of neuronal loss in the entorhinal cortex by H E and TUNEL staining. Neuronal damage was observed in the entorhinal cortex of dCLN ligated 5XFAD, but no neuronal damage was observed in dCLN ligated 5XFAD after FUS-MB. b . Representative merged images of GFAP (green), Aβ (red), and DAPI (blue) immunostaining in hippocampus and entorhinal cortex (left). The isolated images of GFAP to compare the changes of reactive astrocytes (right). Reactive astrocytes were reduced only in entorhinal cortex after FUS-MB. c . Representative merged images of Iba1(red), Aβ (green), and DAPI (blue) immunostaining (left). The isolated images of Iba1 to compare the changes of microglia activation (right). Microglial activation was reduced in the entire brain by FUS-MB.

    Journal: bioRxiv

    Article Title: Improvement of glymphatic-lymphatic drainage of beta-amyloid by focused ultrasound in Alzheimer’s disease model

    doi: 10.1101/2020.01.24.918607

    Figure Lengend Snippet: Effects of brain cells in FUS-MB treated 5XFAD a Representative images of neuronal loss in the entorhinal cortex by H E and TUNEL staining. Neuronal damage was observed in the entorhinal cortex of dCLN ligated 5XFAD, but no neuronal damage was observed in dCLN ligated 5XFAD after FUS-MB. b . Representative merged images of GFAP (green), Aβ (red), and DAPI (blue) immunostaining in hippocampus and entorhinal cortex (left). The isolated images of GFAP to compare the changes of reactive astrocytes (right). Reactive astrocytes were reduced only in entorhinal cortex after FUS-MB. c . Representative merged images of Iba1(red), Aβ (green), and DAPI (blue) immunostaining (left). The isolated images of Iba1 to compare the changes of microglia activation (right). Microglial activation was reduced in the entire brain by FUS-MB.

    Article Snippet: And then brain sections were incubated overnight with the following primary antibodies directed against Aβ (Cell signaling, #8243S), Aβ-6E10 (BioLegend, #803001), GFAP (Cell signaling, #3670S), Iba1 (Abcam, #ab153696), αSMA (Abcam, #ab7817).

    Techniques: TUNEL Assay, Staining, Immunostaining, Isolation, Activation Assay

    Pathological changes in 5XFAD by repeated FUS-MB a. Schematic image of FUS setup and representative image confirming BBB opening with 2% Evans blue after FUS-MB. The BBB opening region was identified around the ipsilateral hippocampus. The red dashed line represents the cut section. b . Design of experiments including 6 repeated FUS-MB, behavioral test and analysis. c . Representative images of Aβ immunostaining in the entorhinal cortex and hippocampus of 5XFAD and age-matched control after 6 repeated FUS-MB. d . Representative images of Aβ immunostaining in serial sections of 5XFAD brain after FUS-MB. The deposition of Aβ decreased in the entire brain by FUS-MB. e . GFAP (green), Iba1 (red), and DAPI (blue) immunostaining images in hippocampus (CA1 and DG) and entorhinal cortex from wild type and 5XFAD mice after 6 repeated FUS to observe changes in astrocyte and microglia activation. Reactive astrocytes were unchanged by FUS-MB, but microglia activation was reduced by FUS-MB.

    Journal: bioRxiv

    Article Title: Improvement of glymphatic-lymphatic drainage of beta-amyloid by focused ultrasound in Alzheimer’s disease model

    doi: 10.1101/2020.01.24.918607

    Figure Lengend Snippet: Pathological changes in 5XFAD by repeated FUS-MB a. Schematic image of FUS setup and representative image confirming BBB opening with 2% Evans blue after FUS-MB. The BBB opening region was identified around the ipsilateral hippocampus. The red dashed line represents the cut section. b . Design of experiments including 6 repeated FUS-MB, behavioral test and analysis. c . Representative images of Aβ immunostaining in the entorhinal cortex and hippocampus of 5XFAD and age-matched control after 6 repeated FUS-MB. d . Representative images of Aβ immunostaining in serial sections of 5XFAD brain after FUS-MB. The deposition of Aβ decreased in the entire brain by FUS-MB. e . GFAP (green), Iba1 (red), and DAPI (blue) immunostaining images in hippocampus (CA1 and DG) and entorhinal cortex from wild type and 5XFAD mice after 6 repeated FUS to observe changes in astrocyte and microglia activation. Reactive astrocytes were unchanged by FUS-MB, but microglia activation was reduced by FUS-MB.

    Article Snippet: And then brain sections were incubated overnight with the following primary antibodies directed against Aβ (Cell signaling, #8243S), Aβ-6E10 (BioLegend, #803001), GFAP (Cell signaling, #3670S), Iba1 (Abcam, #ab153696), αSMA (Abcam, #ab7817).

    Techniques: Immunostaining, Mouse Assay, Activation Assay