anti-neurofilament Search Results


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  • 99
    Thermo Fisher anti neurofilament
    Anti-MHCI antibodies impair synapse elimination at the NMJ in WT mice, while overexpressing MHCI in neurons accelerates synapse elimination (A) Representative NMJs from OX18- and IgG1-injected WT animals at P15. nAChRs labeled with α-btx (red); presynaptic axons and terminals labeled with <t>anti-neurofilament</t> and anti-SV2 (green). Arrows, individual MN axons. Scales, 10μm. (B) Quantification of multiple innervation in IgG1- vs. OX18-injected animals at P15. OX18, 8.8 ± 1.7%; IgG, 3.0 ± 0.8%; *, p = 0.02. OX18 injection, n = 9 animals from 4 litters; IgG1 injection, n = 6 animals from 3 litters. (C) At P7, when synapse elimination is not yet complete, there is a modest but significant decrease in the percentage of muscle fibers that remain multiply-innervated in NSE-D b mice, reflecting the fact that postsynaptic sites more rapidly achieve monoinnervation in MHCI-overexpressing animals. WT, n =3; NSE-D b , n =5; p = 0.012.
    Anti Neurofilament, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti neurofilament
    Case 7, cerebellar pathology: Purkinje neuron with Bielschowsky staining dystrophic granular layer dendrites a and “torpedo” b . Purkinje neurons with ubiquitin staining NCI c , TDP-43 staining NCI d , e and <t>anti-neurofilament</t> antibody staining dystrophic dendrites within the granular layer f . Scale bar 40 μm
    Anti Neurofilament, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti neurofilament
    Epiretinal membrane immunocytochemistry analysis using ( A, B ) a combination of anti-vimentin (green), <t>anti-neurofilament</t> (red), ricin (blue), and Hoescht (white) and ( C , D ) a combination of anti-vimentin (green), anti-ezrin (red), ricin (blue), and Hoescht (white). Notes: Figures ( B and D ) show the four channels separately for Figures ( A and C ), respectively. Mag Bars, ( A , B ): 50 microns; ( C , D ): 100 microns. a,b a Anti-vimentin was used to identify Müller cells (but could also show donor cells); anti-neurofilament was used to identify ganglion cell neurites; ricin was used to identify microglia and macrophages; anti-ezrin was used to identify retinal pigment epithelial cells (red arrows in part C); Hoescht was used to identify nuclei. b Figures are projections of Z-stacks comprising 6 to 10 images taken at 1-micron intervals.
    Anti Neurofilament, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies anti neurofilament
    Treatment of cultured neurons with KPT-350 or knockdown of CRM1 prevents the induction of focal axonal damage and prevents damage in models independent of inflammation ( a ). Representative low and high magnification images of rat hippocampal neurons treated with glutamate (50 μM) and TNFα (200 ng/mL) for 4 hr and stained with <t>neurofilament</t> NFH (red) to identify neuronal processes and SMI-32 (green) to detect damaged neurons with localized swellings. Scale bar = 25μm for low magnification and 15 μm for high magnification ( b ) Quantification of focal axonal damage expressed as percentage of neuronal processes with beading as shown in ( a ), n = 10 neurites per field, 10 fields per experiment, three independent biological replicates. ( c ) Representative confocal images of neurons infected with either control or Xpo1 knockdown lentiviral particles. Xpo1 knockdown was confirmed by staining for CRM1 (blue). Focal axonal damage was evaluated by staining neuronal cultures with neurofilament NFH (red) to identify neuronal processes and SMI-32 (green) to detect damaged neurites in neuronal cultures treated kainic acid. Scale bar = 25 μm for low magnification and 5 μm for high magnification ( d ) Representative still images of mitochondria from videos of cultured neurons stained with the live MitoTracker® Green FM dye; scale bar = 25 μm. ( e ) Quantification of mitochondrial velocity from ( d ), n = > 10 mitochondria per condition in three independent biological replicates. ( f ) Quantification of mitochondrial length from ( d ), n = 150 mitochondria per condition, from three independent experiments. Bar graphs represent mean pixel intensity ± SEM. Statistical significance in: ( b ) was determined using one-way ANOVA with Tukey's test (***p
    Anti Neurofilament, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Developmental Studies Hybridoma Bank anti neurofilament
    Loss of hair cells leads to increased numbers of macrophages in the spiral ganglion. Pou4f3 +/+ and Pou4f3 DTR/+ mice were injected with DT, and cochleae were examined at 1, 3, 7, 14, 28, and 56 d recovery. A , Mid-modiolar frozen section from a DT-treated Pou4f3 +/+ (control) mouse, showing GFP-expressing macrophages (green) and neurons <t>(Neurofilament/TUJ-1,</t> red). B–D , Comparable images taken from mid-modiolar sections of DT-treated Pou4f3 DTR/+ mice at various recovery times. E , Quantification of macrophage numbers per 1000 μm 2 of spiral ganglia (middle region of the cochlea) of control and Pou4f3 DTR/+ mice. Increased numbers of macrophages were observed in mice that had DT-induced hair cell lesions. Notably, macrophage numbers remained elevated as late as 56 d after DT. Data are mean ± SD; n = 3–7 per group. ** p
    Anti Neurofilament, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 93/100, based on 317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology anti neurofilament
    Neurite guidance in the 60, 90, 120 μm ACMFP and control (flat PLGA membrane) groups. ( a ) Confocal images show labeling of DAPI (Cyan), <t>Neurofilament</t> (NF-L; Magenta, marker for neurons and neurites) in the 60, 90, 120 μm ACMFPs and control groups (confocal with DIC microscopy). ( b ). Scale bar: 100 μm.
    Anti Neurofilament, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    AvesLabs anti neurofilament
    Neurite guidance in the 60, 90, 120 μm ACMFP and control (flat PLGA membrane) groups. ( a ) Confocal images show labeling of DAPI (Cyan), <t>Neurofilament</t> (NF-L; Magenta, marker for neurons and neurites) in the 60, 90, 120 μm ACMFPs and control groups (confocal with DIC microscopy). ( b ). Scale bar: 100 μm.
    Anti Neurofilament, supplied by AvesLabs, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Covance anti neurofilament
    Expression of Olig1 in sciatic nerves and cultured Schwann cells Immunofluorescent stainings of transversal (A) and longitudinal (C) tissue sections of sciatic nerves from P7 mice revealed Olig1 immunofluorescence in Remak bundles (arrows), identified by a bundle of unmyelinated (MBP-negative) small diameter axons, and in a subset of myelinating Schwann cells (arrowheads). (B) Remak bundle localization was confirmed by the expression of p75 NTR . (D) A progressive increase of Olig1 mRNA expression levels was detected during peripheral nerve development by qRT–PCR. Data were normalized to the expression of 60s, and values at P0 were set to 1. Each data point represent the mean value of at least eight experimental samples, and the error bars indicate the S.D. (E) In vitro , Olig1 expression was predominantly detected in the cytoplasm of cultured Schwann cells (E, arrows), whereas nuclear staining was significantly weaker (E, arrowheads). NF: <t>neurofilament.</t> Bar: A: 100 μm; B, C, E: 20 μm.
    Anti Neurofilament, supplied by Covance, used in various techniques. Bioz Stars score: 92/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti neurofilament antibody
    Micrographs of SGNs. (A) Bipolar neurons present in cultured SGNs (arrows; magnification, ×100); (B) SGNs stained with <t>anti-neurofilament</t> protein, which is used for the identification of SGNs; green signals can be observed in the membrane (arrow; magnification ×200) rather than in the nuclei. SGNs, spiral ganglion neurons.
    Anti Neurofilament Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad anti neurofilament
    Micrographs of SGNs. (A) Bipolar neurons present in cultured SGNs (arrows; magnification, ×100); (B) SGNs stained with <t>anti-neurofilament</t> protein, which is used for the identification of SGNs; green signals can be observed in the membrane (arrow; magnification ×200) rather than in the nuclei. SGNs, spiral ganglion neurons.
    Anti Neurofilament, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Marque anti neurofilament
    Representative histological features of brainstem GGs from patients 1-4 with representative immunohistochemistry for NF <t>(neurofilament),</t> synaptophysin (SYP), chromogranin (CHR), and VE1 (BRAF), showing large numbers of dysmorphic, irregularly shaped and
    Anti Neurofilament, supplied by Cell Marque, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche anti neurofilament
    Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against <t>neurofilament</t> ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.
    Anti Neurofilament, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Covance anti neurofilament marker
    Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against <t>neurofilament</t> ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.
    Anti Neurofilament Marker, supplied by Covance, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend anti neurofilament anti nf h
    Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against <t>neurofilament</t> ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.
    Anti Neurofilament Anti Nf H, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boster Bio anti neurofilament
    Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against <t>neurofilament</t> ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.
    Anti Neurofilament, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Developmental Studies Hybridoma Bank anti neurofilament 2h3
    Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against <t>neurofilament</t> ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.
    Anti Neurofilament 2h3, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Novocastra anti neurofilament nf200
    Degenerative processes in APP 23 mice. ( A and B ) Staining for acetylcholinesterase in 12-month-old transgenic and control mice, respectively. A local distortion of cholinergic fibers in the plaque vicinity ( A ) compared with normal animals ( B ) can be noted. Neuritic spheroids (arrows) are stained with the <t>neurofilament</t> antibody <t>NF200</t> ( C ). The loss of pyramidal neurons in the vicinity of Aβ deposits in area CA3 is shown in D by toluidine blue staining. (Bars = 50 μm.)
    Anti Neurofilament Nf200, supplied by Novocastra, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Anti-MHCI antibodies impair synapse elimination at the NMJ in WT mice, while overexpressing MHCI in neurons accelerates synapse elimination (A) Representative NMJs from OX18- and IgG1-injected WT animals at P15. nAChRs labeled with α-btx (red); presynaptic axons and terminals labeled with anti-neurofilament and anti-SV2 (green). Arrows, individual MN axons. Scales, 10μm. (B) Quantification of multiple innervation in IgG1- vs. OX18-injected animals at P15. OX18, 8.8 ± 1.7%; IgG, 3.0 ± 0.8%; *, p = 0.02. OX18 injection, n = 9 animals from 4 litters; IgG1 injection, n = 6 animals from 3 litters. (C) At P7, when synapse elimination is not yet complete, there is a modest but significant decrease in the percentage of muscle fibers that remain multiply-innervated in NSE-D b mice, reflecting the fact that postsynaptic sites more rapidly achieve monoinnervation in MHCI-overexpressing animals. WT, n =3; NSE-D b , n =5; p = 0.012.

    Journal: Brain, behavior, and immunity

    Article Title: MHCI promotes developmental synapse elimination and aging-related synapse loss at the vertebrate neuromuscular junction

    doi: 10.1016/j.bbi.2016.01.008

    Figure Lengend Snippet: Anti-MHCI antibodies impair synapse elimination at the NMJ in WT mice, while overexpressing MHCI in neurons accelerates synapse elimination (A) Representative NMJs from OX18- and IgG1-injected WT animals at P15. nAChRs labeled with α-btx (red); presynaptic axons and terminals labeled with anti-neurofilament and anti-SV2 (green). Arrows, individual MN axons. Scales, 10μm. (B) Quantification of multiple innervation in IgG1- vs. OX18-injected animals at P15. OX18, 8.8 ± 1.7%; IgG, 3.0 ± 0.8%; *, p = 0.02. OX18 injection, n = 9 animals from 4 litters; IgG1 injection, n = 6 animals from 3 litters. (C) At P7, when synapse elimination is not yet complete, there is a modest but significant decrease in the percentage of muscle fibers that remain multiply-innervated in NSE-D b mice, reflecting the fact that postsynaptic sites more rapidly achieve monoinnervation in MHCI-overexpressing animals. WT, n =3; NSE-D b , n =5; p = 0.012.

    Article Snippet: Presynaptic MN terminals were visualized by treating muscles for 48-72 hours at 4°C with a primary antibody cocktail consisting of one or both of the following: anti-neurofilament (Invitrogen, 1:500) and anti-SV2 (developed by K. M. Buckley under the auspices of the NICHD, maintained by The University of Iowa Department of Biology, obtained from the Developmental Studies Hybridoma Bank, 1:100).

    Techniques: Mouse Assay, Injection, Labeling

    Persistent multiple innervation at the MHCI-deficient NMJ (A) Confocal images of intact muscle stained with α-btx (red) and anti-neurofilament and anti-SV2 (green). Scales, 10μm. Arrows mark distinct MN axons in P15 and adult. (B) Multiple innervation in confocal images. Multiple innervation at P7, 63.1 ± 1.7% in β2m -/- TAP -/- ( n = 6) vs. 58.5 ± 3.3% in WT ( n = 8), p = 0.28. P15, β2m -/- TAP -/- n = 9, WT n = 8; adult (P29-50), n = 4 per genotype (values, see text). (C) Time course of synapse elimination. Multiple innervation at P0, 83.3 ± 3.1% in β2m -/- TAP -/- vs. 90.2 ± 4.6% in WT, p = 0.13, n = 4 animals per genotype. (B) and (C), **, p = 0.0011; *, p = 0.0116. (D) Multiple innervation in one year old diaphragm. β2m -/- TAP -/- n = 5, WT n = 3. (E) Multiple innervation in mice lacking classical MHCIs. K b-/- , n = 3; D b-/- , n = 6. Gray bars, multiple innervation (mean ± SEM) at the same age in β2m -/- TAP -/- (top) or WT (bottom) diaphragm, from (B). **, p = 0.002; ****, p ≤ 0.0001.

    Journal: Brain, behavior, and immunity

    Article Title: MHCI promotes developmental synapse elimination and aging-related synapse loss at the vertebrate neuromuscular junction

    doi: 10.1016/j.bbi.2016.01.008

    Figure Lengend Snippet: Persistent multiple innervation at the MHCI-deficient NMJ (A) Confocal images of intact muscle stained with α-btx (red) and anti-neurofilament and anti-SV2 (green). Scales, 10μm. Arrows mark distinct MN axons in P15 and adult. (B) Multiple innervation in confocal images. Multiple innervation at P7, 63.1 ± 1.7% in β2m -/- TAP -/- ( n = 6) vs. 58.5 ± 3.3% in WT ( n = 8), p = 0.28. P15, β2m -/- TAP -/- n = 9, WT n = 8; adult (P29-50), n = 4 per genotype (values, see text). (C) Time course of synapse elimination. Multiple innervation at P0, 83.3 ± 3.1% in β2m -/- TAP -/- vs. 90.2 ± 4.6% in WT, p = 0.13, n = 4 animals per genotype. (B) and (C), **, p = 0.0011; *, p = 0.0116. (D) Multiple innervation in one year old diaphragm. β2m -/- TAP -/- n = 5, WT n = 3. (E) Multiple innervation in mice lacking classical MHCIs. K b-/- , n = 3; D b-/- , n = 6. Gray bars, multiple innervation (mean ± SEM) at the same age in β2m -/- TAP -/- (top) or WT (bottom) diaphragm, from (B). **, p = 0.002; ****, p ≤ 0.0001.

    Article Snippet: Presynaptic MN terminals were visualized by treating muscles for 48-72 hours at 4°C with a primary antibody cocktail consisting of one or both of the following: anti-neurofilament (Invitrogen, 1:500) and anti-SV2 (developed by K. M. Buckley under the auspices of the NICHD, maintained by The University of Iowa Department of Biology, obtained from the Developmental Studies Hybridoma Bank, 1:100).

    Techniques: Staining, Mouse Assay

    Case 7, cerebellar pathology: Purkinje neuron with Bielschowsky staining dystrophic granular layer dendrites a and “torpedo” b . Purkinje neurons with ubiquitin staining NCI c , TDP-43 staining NCI d , e and anti-neurofilament antibody staining dystrophic dendrites within the granular layer f . Scale bar 40 μm

    Journal: Journal of neurology

    Article Title: Familial frontotemporal dementia with amyotrophic lateral sclerosis and a shared haplotype on chromosome 9p

    doi: 10.1007/s00415-010-5815-x

    Figure Lengend Snippet: Case 7, cerebellar pathology: Purkinje neuron with Bielschowsky staining dystrophic granular layer dendrites a and “torpedo” b . Purkinje neurons with ubiquitin staining NCI c , TDP-43 staining NCI d , e and anti-neurofilament antibody staining dystrophic dendrites within the granular layer f . Scale bar 40 μm

    Article Snippet: Immuno-histochemistry using dehydrated paraffin wax sections was performed using rabbit anti-human tau (1/500, DAKO), anti-ubiquitin (1/250, DAKO), anti-α-synuclein (1/80, Novocastra), anti-glial fibrillary acidic protein (GFAP) (1/1500, DAKO) and two types of anti-neurofilament (Sigma: 200 kDa, phosphorylated and non-phosphorylated 1/800; ICN Pharmaceuticals: 70 and 200 kDa 1/160) and mouse anti-human monoclonal A-beta (Aβ4 at 1/100, pre-treated with formic acid).

    Techniques: Staining

    Epiretinal membrane immunocytochemistry analysis using ( A, B ) a combination of anti-vimentin (green), anti-neurofilament (red), ricin (blue), and Hoescht (white) and ( C , D ) a combination of anti-vimentin (green), anti-ezrin (red), ricin (blue), and Hoescht (white). Notes: Figures ( B and D ) show the four channels separately for Figures ( A and C ), respectively. Mag Bars, ( A , B ): 50 microns; ( C , D ): 100 microns. a,b a Anti-vimentin was used to identify Müller cells (but could also show donor cells); anti-neurofilament was used to identify ganglion cell neurites; ricin was used to identify microglia and macrophages; anti-ezrin was used to identify retinal pigment epithelial cells (red arrows in part C); Hoescht was used to identify nuclei. b Figures are projections of Z-stacks comprising 6 to 10 images taken at 1-micron intervals.

    Journal: Clinical Ophthalmology (Auckland, N.Z.)

    Article Title: Epiretinal membrane in a subject after transvitreal delivery of palucorcel (CNTO 2476)

    doi: 10.2147/OPTH.S140218

    Figure Lengend Snippet: Epiretinal membrane immunocytochemistry analysis using ( A, B ) a combination of anti-vimentin (green), anti-neurofilament (red), ricin (blue), and Hoescht (white) and ( C , D ) a combination of anti-vimentin (green), anti-ezrin (red), ricin (blue), and Hoescht (white). Notes: Figures ( B and D ) show the four channels separately for Figures ( A and C ), respectively. Mag Bars, ( A , B ): 50 microns; ( C , D ): 100 microns. a,b a Anti-vimentin was used to identify Müller cells (but could also show donor cells); anti-neurofilament was used to identify ganglion cell neurites; ricin was used to identify microglia and macrophages; anti-ezrin was used to identify retinal pigment epithelial cells (red arrows in part C); Hoescht was used to identify nuclei. b Figures are projections of Z-stacks comprising 6 to 10 images taken at 1-micron intervals.

    Article Snippet: The probes used for one piece of tissue included the following: anti-vimentin (1:1,000, Millipore, Temecula, CA, USA) to identify Müller cells, anti-neurofilament (1:500, Abcam, Cambridge, MA, USA) to identify ganglion cell neurites, and biotinylated ricinus communis (1:1,000, 60 kD; Vector Labs, Burlingame, CA, USA) to identify microglia and macrophages.

    Techniques: Immunocytochemistry

    The abnormal architecture of the neuromuscular junction (NMJ) with axonal denervation in conditional knockout (cKO) mice. (A) Fluorescent images of NMJ in soleus muscle (SOL) from mice at age 6 and 16 weeks. Immunofluorescence assay with α‐bungarotoxin (BTX) [red, for acetylcholine receptor (AChR)] and anti‐synaptophysin (Syn) and anti‐neurofilament (NF) antibodies (both green, for nerve terminals) used for analysis of NMJ area and the patterns of axonal innervation. Scale bar: 10 μm. (B) Quantification of NMJ area in soleus muscle in mice at age 6 and 16 weeks. The area of BTX‐labelled AChR represents the NMJ area. n = 5 for 6 week and n = 7 for 16 week mice. The number in each bar indicates the number of examined NMJs. (C) Patterns of axonal innervation in mice at age 16 weeks. Three NMJ innervation patterns are indicated, including innervated (well overlapping of AChR and nerve terminals), intermediate (nerve terminal adjacent to AChR without overlapping), and denervated (no nerve terminal adjacent or overlapped to AChR). n = 3 for each group. Scale bar: 10 μm. (D) Proportion of innervated, intermediate, and denervated endplates in soleus muscle from mice at age 16 weeks. (E) Quantification of NMJ area in gastrocnemius muscle (GAS) from mice at age 16 weeks and (F) proportion of denervated endplates in gastrocnemius muscle. n = 3 for each group. (G) Pathological and motor functional results in cKO mice at age 6 and 16 weeks. Data are mean ± SEM. Panels D and F: by Student's t ‐test; panels B and E: one‐way analysis of variance. * P

    Journal: Journal of Cachexia, Sarcopenia and Muscle

    Article Title: Muscle‐restricted nuclear receptor interaction protein knockout causes motor neuron degeneration through down‐regulation of myogenin at the neuromuscular junction) Muscle‐restricted nuclear receptor interaction protein knockout causes motor neuron degeneration through down‐regulation of myogenin at the neuromuscular junction

    doi: 10.1002/jcsm.12299

    Figure Lengend Snippet: The abnormal architecture of the neuromuscular junction (NMJ) with axonal denervation in conditional knockout (cKO) mice. (A) Fluorescent images of NMJ in soleus muscle (SOL) from mice at age 6 and 16 weeks. Immunofluorescence assay with α‐bungarotoxin (BTX) [red, for acetylcholine receptor (AChR)] and anti‐synaptophysin (Syn) and anti‐neurofilament (NF) antibodies (both green, for nerve terminals) used for analysis of NMJ area and the patterns of axonal innervation. Scale bar: 10 μm. (B) Quantification of NMJ area in soleus muscle in mice at age 6 and 16 weeks. The area of BTX‐labelled AChR represents the NMJ area. n = 5 for 6 week and n = 7 for 16 week mice. The number in each bar indicates the number of examined NMJs. (C) Patterns of axonal innervation in mice at age 16 weeks. Three NMJ innervation patterns are indicated, including innervated (well overlapping of AChR and nerve terminals), intermediate (nerve terminal adjacent to AChR without overlapping), and denervated (no nerve terminal adjacent or overlapped to AChR). n = 3 for each group. Scale bar: 10 μm. (D) Proportion of innervated, intermediate, and denervated endplates in soleus muscle from mice at age 16 weeks. (E) Quantification of NMJ area in gastrocnemius muscle (GAS) from mice at age 16 weeks and (F) proportion of denervated endplates in gastrocnemius muscle. n = 3 for each group. (G) Pathological and motor functional results in cKO mice at age 6 and 16 weeks. Data are mean ± SEM. Panels D and F: by Student's t ‐test; panels B and E: one‐way analysis of variance. * P

    Article Snippet: Next, soleus was blocked in 2% bovine serum albumin and incubated with the antibodies anti‐neurofilament (Abcam, Cambridge, MA, USA 1:500 dilution) and anti‐synaptophysin (Abcam, 1:200 dilution) overnight at 4°C.

    Techniques: Knock-Out, Mouse Assay, Immunofluorescence, Functional Assay

    Intramuscular myogenin gene transfer rescues phenotypes of conditional knockout (cKO) mice. (A) Analysis of mice at age 9 and 16 weeks after adenovirus encoding myogenin (Ad‐myogenin) and Ad‐shLuc (control) treatment by intramuscular (i.m.) injection into bilateral gastrocnemius muscle from cKO mice at age 6 weeks. (B) Age 9 weeks: western blot analysis of myogenin and nuclear receptor interaction protein (NRIP) levels in gastrocnemius muscle from cKO mice infected with Ad‐myogenin ( n = 4) or control ( n = 3), with tubulin as an internal control. (C) Age 9 weeks: immunofluorescence assay of myogenin expression (green) in gastrocnemius muscle from cKO mice with Ad‐myogenin ( n = 6) or Ad‐shLuc ( n = 6) treatment. Scale bar, 20 μm. (D) Age 9 weeks: quantification of immunohistochemical staining of proportion of oxidative fibres and oxidative fibre cross‐section area (CSA) for slow myosin in soleus muscle from cKO mice with Ad‐myogenin treatment ( n = 6) or control ( n = 6). (E) Age 9 weeks: proportion of succinate dehydrogenase (SDH)‐stained myofibers in soleus muscle from cKO mice with Ad‐myogenin treatment ( n = 6) or control ( n = 4). (F) Immunofluorescence assay with α‐bungarotoxin (BTX) (red) and anti‐synaptophysin (Syn) and anti‐neurofilament (NF) antibodies (green) for analysis of neuromuscular junction (NMJ) area and patterns of axonal innervation in gastrocnemius muscle from cKO mice at age 16 weeks. Right: quantification of NMJ area in gastrocnemius muscle from cKO mice at age 9 and 16 weeks. The number in each bar indicates the number of examined NMJs. (G) Proportion of denervated endplates in gastrocnemius muscle from cKO mice. (H) Sections (30 μm) of lumbar spinal cord were stained for choline acetyltransferase and neuronal DNA binding protein for α‐motor neuron labelling. The number of α‐motor neurons per spinal anterior horn was measured. (G and H): the N in each bar indicates the number of mice. Data are mean ± SEM. Panels C–E: by Student's t ‐test. Panels F–H: one‐way analysis of variance. * P

    Journal: Journal of Cachexia, Sarcopenia and Muscle

    Article Title: Muscle‐restricted nuclear receptor interaction protein knockout causes motor neuron degeneration through down‐regulation of myogenin at the neuromuscular junction) Muscle‐restricted nuclear receptor interaction protein knockout causes motor neuron degeneration through down‐regulation of myogenin at the neuromuscular junction

    doi: 10.1002/jcsm.12299

    Figure Lengend Snippet: Intramuscular myogenin gene transfer rescues phenotypes of conditional knockout (cKO) mice. (A) Analysis of mice at age 9 and 16 weeks after adenovirus encoding myogenin (Ad‐myogenin) and Ad‐shLuc (control) treatment by intramuscular (i.m.) injection into bilateral gastrocnemius muscle from cKO mice at age 6 weeks. (B) Age 9 weeks: western blot analysis of myogenin and nuclear receptor interaction protein (NRIP) levels in gastrocnemius muscle from cKO mice infected with Ad‐myogenin ( n = 4) or control ( n = 3), with tubulin as an internal control. (C) Age 9 weeks: immunofluorescence assay of myogenin expression (green) in gastrocnemius muscle from cKO mice with Ad‐myogenin ( n = 6) or Ad‐shLuc ( n = 6) treatment. Scale bar, 20 μm. (D) Age 9 weeks: quantification of immunohistochemical staining of proportion of oxidative fibres and oxidative fibre cross‐section area (CSA) for slow myosin in soleus muscle from cKO mice with Ad‐myogenin treatment ( n = 6) or control ( n = 6). (E) Age 9 weeks: proportion of succinate dehydrogenase (SDH)‐stained myofibers in soleus muscle from cKO mice with Ad‐myogenin treatment ( n = 6) or control ( n = 4). (F) Immunofluorescence assay with α‐bungarotoxin (BTX) (red) and anti‐synaptophysin (Syn) and anti‐neurofilament (NF) antibodies (green) for analysis of neuromuscular junction (NMJ) area and patterns of axonal innervation in gastrocnemius muscle from cKO mice at age 16 weeks. Right: quantification of NMJ area in gastrocnemius muscle from cKO mice at age 9 and 16 weeks. The number in each bar indicates the number of examined NMJs. (G) Proportion of denervated endplates in gastrocnemius muscle from cKO mice. (H) Sections (30 μm) of lumbar spinal cord were stained for choline acetyltransferase and neuronal DNA binding protein for α‐motor neuron labelling. The number of α‐motor neurons per spinal anterior horn was measured. (G and H): the N in each bar indicates the number of mice. Data are mean ± SEM. Panels C–E: by Student's t ‐test. Panels F–H: one‐way analysis of variance. * P

    Article Snippet: Next, soleus was blocked in 2% bovine serum albumin and incubated with the antibodies anti‐neurofilament (Abcam, Cambridge, MA, USA 1:500 dilution) and anti‐synaptophysin (Abcam, 1:200 dilution) overnight at 4°C.

    Techniques: Knock-Out, Mouse Assay, Injection, Western Blot, Infection, Immunofluorescence, Expressing, Immunohistochemistry, Staining, Binding Assay

    Peripheral SGN connectivity is normal in Npr2 mutant mice. ( A,B ) Cochlear innervation was visualized by neurofilament immunostaining of P18 whole cochleae. Visual inspection revealed no obvious difference in the peripheral pattern of connections between wild-type control (Ctl, n = 2) (A) and Npr2 mutant (Mut, n = 2) (B) animals. ( C,D ) SGN projections in the P14 cochlea were labeled by crossing Neurog1-CreER T2 to the AI14:tdTomato reporter strain, which enables visualization of a subset of SGNs along the length of the cochlea. Individual SGNs of control animals (Ctl, n = 3 heterozygotes) (C) show neatly organized projections within the cochlea. Individual SGN processes of Npr2 mutant (Mut, n = 5) cochleae (D) showed no qualitative differences compared to controls. Note that the degree of labeling can vary slightly independent of genotype, due to fluctuations in Cre activity. ( E ) Electron micrograph of a transverse section of myelinated SGN axons in the eighth nerve in a control P21 animal. ( F ) Similar electron micrograph of the eighth nerve of an Npr2 mutant at P21 shows normal axonal diameters and normal myelination. ( G ) The g-ratio of Npr2 mutants did not differ from controls ( P = 0.87, Student's t-test) and was near optimal. Scale bars in A, 50 µm; C, 2 µm.

    Journal: PLoS Genetics

    Article Title: Mutation of Npr2 Leads to Blurred Tonotopic Organization of Central Auditory Circuits in Mice

    doi: 10.1371/journal.pgen.1004823

    Figure Lengend Snippet: Peripheral SGN connectivity is normal in Npr2 mutant mice. ( A,B ) Cochlear innervation was visualized by neurofilament immunostaining of P18 whole cochleae. Visual inspection revealed no obvious difference in the peripheral pattern of connections between wild-type control (Ctl, n = 2) (A) and Npr2 mutant (Mut, n = 2) (B) animals. ( C,D ) SGN projections in the P14 cochlea were labeled by crossing Neurog1-CreER T2 to the AI14:tdTomato reporter strain, which enables visualization of a subset of SGNs along the length of the cochlea. Individual SGNs of control animals (Ctl, n = 3 heterozygotes) (C) show neatly organized projections within the cochlea. Individual SGN processes of Npr2 mutant (Mut, n = 5) cochleae (D) showed no qualitative differences compared to controls. Note that the degree of labeling can vary slightly independent of genotype, due to fluctuations in Cre activity. ( E ) Electron micrograph of a transverse section of myelinated SGN axons in the eighth nerve in a control P21 animal. ( F ) Similar electron micrograph of the eighth nerve of an Npr2 mutant at P21 shows normal axonal diameters and normal myelination. ( G ) The g-ratio of Npr2 mutants did not differ from controls ( P = 0.87, Student's t-test) and was near optimal. Scale bars in A, 50 µm; C, 2 µm.

    Article Snippet: Visualization of cochlear afferents To examine the overall pattern of peripheral projections in the cochlea, wild-type (n = 2) or Npr2 mutant (n = 2) animals were perfused transcardially with 4% PFA in PBS, and the cochleae were dissected out and fixed overnight at 4°C, then subjected to whole-mount immunofluorescence with chick anti-Neurofilament antibody (1∶1000, Abcam), using Alexa488-conjugated goat anti-chick secondary antibody (1∶1000, Life Technologies).

    Techniques: Mutagenesis, Mouse Assay, Immunostaining, CTL Assay, Labeling, Activity Assay

    MP201 treatment reduces axonal loss in optic neuritis. Neurofilament staining was used to evaluate axonal loss in sections of optic nerves isolated at day 42 postimmunization. (a) The optical density of neurofilament staining calculated from three equal-sized fields from each optic nerve shows a significant decrease ( ∗∗∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Mitochondrial Uncoupler Prodrug of 2,4-Dinitrophenol, MP201, Prevents Neuronal Damage and Preserves Vision in Experimental Optic Neuritis

    doi: 10.1155/2017/7180632

    Figure Lengend Snippet: MP201 treatment reduces axonal loss in optic neuritis. Neurofilament staining was used to evaluate axonal loss in sections of optic nerves isolated at day 42 postimmunization. (a) The optical density of neurofilament staining calculated from three equal-sized fields from each optic nerve shows a significant decrease ( ∗∗∗ p

    Article Snippet: Specimens were then incubated in rabbit anti-neurofilament antibody 1 : 100 (Abcam, Cambridge, MA, USA) at 4°C overnight and then washed three times with PBS and incubated with anti-rabbit secondary antibody (Vectastain Elite ABC Rabbit kit) for 30 min at 37°C.

    Techniques: Staining, Isolation

    Characterisation of neuronal differentiation through the stages of neurite outgrowth from neurospheres over 20 days in 2D culture. Expression over time of nestin (A–D), neurofilament-L (E–H), neurofilament-M (I–L), neurofilament-H (M–P), and TUJ-1 (Q–T). Scale bars: (A–P): 50 μm, (Q–T): 100 μm.

    Journal: Neurochemistry International

    Article Title: A robust and reproducible human pluripotent stem cell derived model of neurite outgrowth in a three-dimensional culture system and its application to study neurite inhibition

    doi: 10.1016/j.neuint.2016.12.009

    Figure Lengend Snippet: Characterisation of neuronal differentiation through the stages of neurite outgrowth from neurospheres over 20 days in 2D culture. Expression over time of nestin (A–D), neurofilament-L (E–H), neurofilament-M (I–L), neurofilament-H (M–P), and TUJ-1 (Q–T). Scale bars: (A–P): 50 μm, (Q–T): 100 μm.

    Article Snippet: 2D cell culture samples were then incubated with primary antibodies (anti-β—III—tubulin (Cambridge bioscience, Cambridge, UK), anti-nestin (Abcam, Cambridge, UK), anti-Neurofilament-H (Abcam), anti-neurofilament-M (Sigma-Aldrich) or anti-neurofilment-L (Sigma-Aldrich)) on ice for 60 min, whereas cells on Alvetex® scaffolds were incubated for 120 min before being washed 3 times in blocking buffer.

    Techniques: Expressing

    Treatment of cultured neurons with KPT-350 or knockdown of CRM1 prevents the induction of focal axonal damage and prevents damage in models independent of inflammation ( a ). Representative low and high magnification images of rat hippocampal neurons treated with glutamate (50 μM) and TNFα (200 ng/mL) for 4 hr and stained with neurofilament NFH (red) to identify neuronal processes and SMI-32 (green) to detect damaged neurons with localized swellings. Scale bar = 25μm for low magnification and 15 μm for high magnification ( b ) Quantification of focal axonal damage expressed as percentage of neuronal processes with beading as shown in ( a ), n = 10 neurites per field, 10 fields per experiment, three independent biological replicates. ( c ) Representative confocal images of neurons infected with either control or Xpo1 knockdown lentiviral particles. Xpo1 knockdown was confirmed by staining for CRM1 (blue). Focal axonal damage was evaluated by staining neuronal cultures with neurofilament NFH (red) to identify neuronal processes and SMI-32 (green) to detect damaged neurites in neuronal cultures treated kainic acid. Scale bar = 25 μm for low magnification and 5 μm for high magnification ( d ) Representative still images of mitochondria from videos of cultured neurons stained with the live MitoTracker® Green FM dye; scale bar = 25 μm. ( e ) Quantification of mitochondrial velocity from ( d ), n = > 10 mitochondria per condition in three independent biological replicates. ( f ) Quantification of mitochondrial length from ( d ), n = 150 mitochondria per condition, from three independent experiments. Bar graphs represent mean pixel intensity ± SEM. Statistical significance in: ( b ) was determined using one-way ANOVA with Tukey's test (***p

    Journal: Nature neuroscience

    Article Title: Selective inhibitors of nuclear export avert progression in preclinical models of inflammatory demyelination

    doi: 10.1038/nn.3953

    Figure Lengend Snippet: Treatment of cultured neurons with KPT-350 or knockdown of CRM1 prevents the induction of focal axonal damage and prevents damage in models independent of inflammation ( a ). Representative low and high magnification images of rat hippocampal neurons treated with glutamate (50 μM) and TNFα (200 ng/mL) for 4 hr and stained with neurofilament NFH (red) to identify neuronal processes and SMI-32 (green) to detect damaged neurons with localized swellings. Scale bar = 25μm for low magnification and 15 μm for high magnification ( b ) Quantification of focal axonal damage expressed as percentage of neuronal processes with beading as shown in ( a ), n = 10 neurites per field, 10 fields per experiment, three independent biological replicates. ( c ) Representative confocal images of neurons infected with either control or Xpo1 knockdown lentiviral particles. Xpo1 knockdown was confirmed by staining for CRM1 (blue). Focal axonal damage was evaluated by staining neuronal cultures with neurofilament NFH (red) to identify neuronal processes and SMI-32 (green) to detect damaged neurites in neuronal cultures treated kainic acid. Scale bar = 25 μm for low magnification and 5 μm for high magnification ( d ) Representative still images of mitochondria from videos of cultured neurons stained with the live MitoTracker® Green FM dye; scale bar = 25 μm. ( e ) Quantification of mitochondrial velocity from ( d ), n = > 10 mitochondria per condition in three independent biological replicates. ( f ) Quantification of mitochondrial length from ( d ), n = 150 mitochondria per condition, from three independent experiments. Bar graphs represent mean pixel intensity ± SEM. Statistical significance in: ( b ) was determined using one-way ANOVA with Tukey's test (***p

    Article Snippet: Slides were incubated overnight with antibodies for anti-MBP (Dako, cat no: 1: 1000), anti-neurofilament (Dako, 1:2000), anti-CD45 (Dako, 1:800), anti-CRM1 (Santa Cruz, cat no: sc-5595, 1:100), anti-NeuN (Chemicon, cat no: MAB377, 1:100).

    Techniques: Cell Culture, Staining, Infection

    Expression of CRM1 in MS gray and white matter lesion areas All pictures are derived from the same leukocortical MS lesion. ( a ) Immunohistochemical staining of a leukocortical MS lesion, which was characterized by staining for myelin basic protein (MBP), NeuN for neurons and CD68 for monocytes/microglia. The dotted line indicates the border between grey and white matter. Note the loss of myelin and the presence of myelin-laden phagocytes at the edges of the lesion (insert in the MBP staining panel; scale bar = 25 μm). CD68 positive cells were detected in gray and white matter, with higher numbers in the white matter part of the lesion. ( b ) The left panel depicts a perivascular infiltrate in the white matter part of the lesion and the arrow indicates the numerous C68+ positive cells within the infiltrate as well as in the parenchyma. Damaged neurons were identified by somatic staining for neurofilament. ( c ) The fluorescent images show at higher magnification the large inflammatory infiltrate located at the border between white and gray matter indicated by an arrow in ( b ). Numerous CD45 positive leukocytes (green) express CRM1 (red). Nuclei were counterstained with DAPI (blue); scale bar = 100 μm. Low ( d ) and high ( e ) magnification confocal images of lesional areas of MS gray matter and normal appearing gray matter stained for NeuN (green) to identify neurons and CRM1 (red). Scale bar = 40 μm for ( d ) and 15 μm for ( e ).( f ) Western blot analysis of protein extracts of post-mortem human gray matter tissue prepared from non-neurological controls (n = 5) and MS patients (n = 8) which were probed with antibodies specific for CRM1, ACTIN was used as a loading control. Blots were cropped and full length images are presented in Supplementary Figure 11 .

    Journal: Nature neuroscience

    Article Title: Selective inhibitors of nuclear export avert progression in preclinical models of inflammatory demyelination

    doi: 10.1038/nn.3953

    Figure Lengend Snippet: Expression of CRM1 in MS gray and white matter lesion areas All pictures are derived from the same leukocortical MS lesion. ( a ) Immunohistochemical staining of a leukocortical MS lesion, which was characterized by staining for myelin basic protein (MBP), NeuN for neurons and CD68 for monocytes/microglia. The dotted line indicates the border between grey and white matter. Note the loss of myelin and the presence of myelin-laden phagocytes at the edges of the lesion (insert in the MBP staining panel; scale bar = 25 μm). CD68 positive cells were detected in gray and white matter, with higher numbers in the white matter part of the lesion. ( b ) The left panel depicts a perivascular infiltrate in the white matter part of the lesion and the arrow indicates the numerous C68+ positive cells within the infiltrate as well as in the parenchyma. Damaged neurons were identified by somatic staining for neurofilament. ( c ) The fluorescent images show at higher magnification the large inflammatory infiltrate located at the border between white and gray matter indicated by an arrow in ( b ). Numerous CD45 positive leukocytes (green) express CRM1 (red). Nuclei were counterstained with DAPI (blue); scale bar = 100 μm. Low ( d ) and high ( e ) magnification confocal images of lesional areas of MS gray matter and normal appearing gray matter stained for NeuN (green) to identify neurons and CRM1 (red). Scale bar = 40 μm for ( d ) and 15 μm for ( e ).( f ) Western blot analysis of protein extracts of post-mortem human gray matter tissue prepared from non-neurological controls (n = 5) and MS patients (n = 8) which were probed with antibodies specific for CRM1, ACTIN was used as a loading control. Blots were cropped and full length images are presented in Supplementary Figure 11 .

    Article Snippet: Slides were incubated overnight with antibodies for anti-MBP (Dako, cat no: 1: 1000), anti-neurofilament (Dako, 1:2000), anti-CD45 (Dako, 1:800), anti-CRM1 (Santa Cruz, cat no: sc-5595, 1:100), anti-NeuN (Chemicon, cat no: MAB377, 1:100).

    Techniques: Expressing, Mass Spectrometry, Derivative Assay, Immunohistochemistry, Staining, Western Blot

    Loss of hair cells leads to increased numbers of macrophages in the spiral ganglion. Pou4f3 +/+ and Pou4f3 DTR/+ mice were injected with DT, and cochleae were examined at 1, 3, 7, 14, 28, and 56 d recovery. A , Mid-modiolar frozen section from a DT-treated Pou4f3 +/+ (control) mouse, showing GFP-expressing macrophages (green) and neurons (Neurofilament/TUJ-1, red). B–D , Comparable images taken from mid-modiolar sections of DT-treated Pou4f3 DTR/+ mice at various recovery times. E , Quantification of macrophage numbers per 1000 μm 2 of spiral ganglia (middle region of the cochlea) of control and Pou4f3 DTR/+ mice. Increased numbers of macrophages were observed in mice that had DT-induced hair cell lesions. Notably, macrophage numbers remained elevated as late as 56 d after DT. Data are mean ± SD; n = 3–7 per group. ** p

    Journal: The Journal of Neuroscience

    Article Title: Fractalkine Signaling Regulates Macrophage Recruitment into the Cochlea and Promotes the Survival of Spiral Ganglion Neurons after Selective Hair Cell Lesion

    doi: 10.1523/JNEUROSCI.2325-15.2015

    Figure Lengend Snippet: Loss of hair cells leads to increased numbers of macrophages in the spiral ganglion. Pou4f3 +/+ and Pou4f3 DTR/+ mice were injected with DT, and cochleae were examined at 1, 3, 7, 14, 28, and 56 d recovery. A , Mid-modiolar frozen section from a DT-treated Pou4f3 +/+ (control) mouse, showing GFP-expressing macrophages (green) and neurons (Neurofilament/TUJ-1, red). B–D , Comparable images taken from mid-modiolar sections of DT-treated Pou4f3 DTR/+ mice at various recovery times. E , Quantification of macrophage numbers per 1000 μm 2 of spiral ganglia (middle region of the cochlea) of control and Pou4f3 DTR/+ mice. Increased numbers of macrophages were observed in mice that had DT-induced hair cell lesions. Notably, macrophage numbers remained elevated as late as 56 d after DT. Data are mean ± SD; n = 3–7 per group. ** p

    Article Snippet: Neuronal peripheral processes and ganglion cell bodies were labeled using a combination of anti-Neurofilament (mouse monoclonal, clone #2H3, Developmental Studies Hybridoma Bank, University of Iowa, 1:100) and anti-β-III tubulin antibodies (mouse monoclonal, catalog #MMS-435P, Covance, 1:500).

    Techniques: Mouse Assay, Injection, Expressing

    Loss of hair cells is sufficient to recruit macrophages toward the cochlear sensory epithelium. Pou4f3 +/+ (control) and Pou4f3 DTR/+ mice received a single DT injection, and cochleae were examined at 1, 3, 7, 14, 28, and 56 d recovery. A–D , Cochlear whole mounts showing GFP-labeled macrophages (green) and neurons (Neurofilament and β-III tubulin, red) demonstrate increased numbers of macrophages within the sensory regions of lesioned (e.g., Pou4f3 DTR/+ ) mice ( B – D ), compared with controls ( A ). Elevated macrophage numbers were first observed at 3 d after DT. E , The 3D renderings of confocal image stacks of lesioned cochleae show that macrophages (green) are typically located below the basilar membrane among the tympanic mesothelial cells and in the osseous spiral lamina. F , Quantification of macrophages per 100 μm of sensory epithelium. Macrophage numbers near the sensory epithelium peak at 14 dpi and diminish by 56 dpi. G , The area in the sensory epithelium (white box) of the cochlea from which macrophages were counted. H , I , The 3D renderings obtained from confocal image stacks of lesioned cochleae show that macrophages (green) are often in close contact with neurons (red) crossing the tunnel of Corti (white arrows) and extended processes toward hair cells (position shown with white arrowheads). Data are mean ± SD; n = 6 per group. dpi, days post injection; NF, neurofilament; IHC, inner hair cell; OHC; outer hair cell; ISB; inner spiral bundle; ToC; tunnel of Corti Statistical significance was computed using two-way ANOVA followed by Tukey's multiple comparison tests. ** p

    Journal: The Journal of Neuroscience

    Article Title: Fractalkine Signaling Regulates Macrophage Recruitment into the Cochlea and Promotes the Survival of Spiral Ganglion Neurons after Selective Hair Cell Lesion

    doi: 10.1523/JNEUROSCI.2325-15.2015

    Figure Lengend Snippet: Loss of hair cells is sufficient to recruit macrophages toward the cochlear sensory epithelium. Pou4f3 +/+ (control) and Pou4f3 DTR/+ mice received a single DT injection, and cochleae were examined at 1, 3, 7, 14, 28, and 56 d recovery. A–D , Cochlear whole mounts showing GFP-labeled macrophages (green) and neurons (Neurofilament and β-III tubulin, red) demonstrate increased numbers of macrophages within the sensory regions of lesioned (e.g., Pou4f3 DTR/+ ) mice ( B – D ), compared with controls ( A ). Elevated macrophage numbers were first observed at 3 d after DT. E , The 3D renderings of confocal image stacks of lesioned cochleae show that macrophages (green) are typically located below the basilar membrane among the tympanic mesothelial cells and in the osseous spiral lamina. F , Quantification of macrophages per 100 μm of sensory epithelium. Macrophage numbers near the sensory epithelium peak at 14 dpi and diminish by 56 dpi. G , The area in the sensory epithelium (white box) of the cochlea from which macrophages were counted. H , I , The 3D renderings obtained from confocal image stacks of lesioned cochleae show that macrophages (green) are often in close contact with neurons (red) crossing the tunnel of Corti (white arrows) and extended processes toward hair cells (position shown with white arrowheads). Data are mean ± SD; n = 6 per group. dpi, days post injection; NF, neurofilament; IHC, inner hair cell; OHC; outer hair cell; ISB; inner spiral bundle; ToC; tunnel of Corti Statistical significance was computed using two-way ANOVA followed by Tukey's multiple comparison tests. ** p

    Article Snippet: Neuronal peripheral processes and ganglion cell bodies were labeled using a combination of anti-Neurofilament (mouse monoclonal, clone #2H3, Developmental Studies Hybridoma Bank, University of Iowa, 1:100) and anti-β-III tubulin antibodies (mouse monoclonal, catalog #MMS-435P, Covance, 1:500).

    Techniques: Mouse Assay, Injection, Labeling, Immunohistochemistry

    Cochlear supporting cells and SGNs are not affected by DT-mediated hair cell ablation. Top row, Cochlear whole mounts immunolabeled for SOX2 (green, supporting cells) from ( A ) Pou4f3 +/+ and Pou4f3 DTR/+ mice at ( B ) 7 d, ( C ) 14 d, and ( D ) 56 d after DT injection. We observed no evidence for the loss of supporting cells, despite the complete ablation of both inner and outer hair cells. Supporting cell nuclei change their shape and frequently redistribute after hair cell loss. E , Average number of SOX2-positive supporting cells at various times after DT injection. Bottom row, Mid-modiolar sections labeled for neurofilament and β-III tubulin (red, neurons) from ( F ) Pou4f3 +/+ and ( G–I ) Pou4f3 DTR/+ mice at 7, 14, and 56 d following DT injection. No loss of spiral ganglion cell bodies was evident at any survival time. J , Average SGN density from the middle region of the cochlea at various survival times. Data are mean ± SD; n = 4–6 per group. Scale bar, 30 μm.

    Journal: The Journal of Neuroscience

    Article Title: Fractalkine Signaling Regulates Macrophage Recruitment into the Cochlea and Promotes the Survival of Spiral Ganglion Neurons after Selective Hair Cell Lesion

    doi: 10.1523/JNEUROSCI.2325-15.2015

    Figure Lengend Snippet: Cochlear supporting cells and SGNs are not affected by DT-mediated hair cell ablation. Top row, Cochlear whole mounts immunolabeled for SOX2 (green, supporting cells) from ( A ) Pou4f3 +/+ and Pou4f3 DTR/+ mice at ( B ) 7 d, ( C ) 14 d, and ( D ) 56 d after DT injection. We observed no evidence for the loss of supporting cells, despite the complete ablation of both inner and outer hair cells. Supporting cell nuclei change their shape and frequently redistribute after hair cell loss. E , Average number of SOX2-positive supporting cells at various times after DT injection. Bottom row, Mid-modiolar sections labeled for neurofilament and β-III tubulin (red, neurons) from ( F ) Pou4f3 +/+ and ( G–I ) Pou4f3 DTR/+ mice at 7, 14, and 56 d following DT injection. No loss of spiral ganglion cell bodies was evident at any survival time. J , Average SGN density from the middle region of the cochlea at various survival times. Data are mean ± SD; n = 4–6 per group. Scale bar, 30 μm.

    Article Snippet: Neuronal peripheral processes and ganglion cell bodies were labeled using a combination of anti-Neurofilament (mouse monoclonal, clone #2H3, Developmental Studies Hybridoma Bank, University of Iowa, 1:100) and anti-β-III tubulin antibodies (mouse monoclonal, catalog #MMS-435P, Covance, 1:500).

    Techniques: Immunolabeling, Mouse Assay, Injection, Labeling

    Genetic deletion of CX 3 CR1 results in significant loss of SGNs after hair cell ablation. Cochlear mid-modiolar sections labeled for neurofilament (neurons, red) revealed significant loss of spiral ganglion cell bodies in the CX 3 CR1 −/− mice ( B′ ), compared with CX 3 CR1 +/− ( A′ ) mice, at 56 d after DT-induced hair cell lesion. We observed no evidence for SGN loss in Pou4f3 +/+ mice that were either CX 3 CR1 −/− ( B ) or CX 3 CR1 +/− ( A ). C , Quantitative data on SGN density (per 1000 μm 2 ) for the four genotypes. D , E , Plastic sections from the basal cochlear turn of Pou4f3 +/+ mice at 56 d after DT injection. Both CX 3 CR1 +/− ( D ) and CX 3 CR1 −/− ( E ) possessed normal numbers of SGNs. D′ , E′ , Plastic sections from the basal cochlear turn of Pou4f3 DTR/+ mice at 56 d after DT injection. Spiral ganglia of CX 3 CR1 +/− mice ( D′ ) contained normal numbers of auditory neurons after hair cell lesions, whereas the ganglia of CX 3 CR1 −/− mice ( E′ ) showed diminished neuronal survival. F , Density of SGNs (per 1000 μm 2 ) from the basal cochlear turn at all time points after DT-induced hair cell lesion. Data are mean ± SD; n = 5–10 per group. dpi, days post-DT injection. Statistical significance was computed using two-way ANOVA followed by Tukey's multiple comparison post hoc tests. * p

    Journal: The Journal of Neuroscience

    Article Title: Fractalkine Signaling Regulates Macrophage Recruitment into the Cochlea and Promotes the Survival of Spiral Ganglion Neurons after Selective Hair Cell Lesion

    doi: 10.1523/JNEUROSCI.2325-15.2015

    Figure Lengend Snippet: Genetic deletion of CX 3 CR1 results in significant loss of SGNs after hair cell ablation. Cochlear mid-modiolar sections labeled for neurofilament (neurons, red) revealed significant loss of spiral ganglion cell bodies in the CX 3 CR1 −/− mice ( B′ ), compared with CX 3 CR1 +/− ( A′ ) mice, at 56 d after DT-induced hair cell lesion. We observed no evidence for SGN loss in Pou4f3 +/+ mice that were either CX 3 CR1 −/− ( B ) or CX 3 CR1 +/− ( A ). C , Quantitative data on SGN density (per 1000 μm 2 ) for the four genotypes. D , E , Plastic sections from the basal cochlear turn of Pou4f3 +/+ mice at 56 d after DT injection. Both CX 3 CR1 +/− ( D ) and CX 3 CR1 −/− ( E ) possessed normal numbers of SGNs. D′ , E′ , Plastic sections from the basal cochlear turn of Pou4f3 DTR/+ mice at 56 d after DT injection. Spiral ganglia of CX 3 CR1 +/− mice ( D′ ) contained normal numbers of auditory neurons after hair cell lesions, whereas the ganglia of CX 3 CR1 −/− mice ( E′ ) showed diminished neuronal survival. F , Density of SGNs (per 1000 μm 2 ) from the basal cochlear turn at all time points after DT-induced hair cell lesion. Data are mean ± SD; n = 5–10 per group. dpi, days post-DT injection. Statistical significance was computed using two-way ANOVA followed by Tukey's multiple comparison post hoc tests. * p

    Article Snippet: Neuronal peripheral processes and ganglion cell bodies were labeled using a combination of anti-Neurofilament (mouse monoclonal, clone #2H3, Developmental Studies Hybridoma Bank, University of Iowa, 1:100) and anti-β-III tubulin antibodies (mouse monoclonal, catalog #MMS-435P, Covance, 1:500).

    Techniques: Labeling, Mouse Assay, Injection

    Genetic deletion of CX 3 CR1 led to a reduction in macrophages near the sensory epithelium after hair cell ablation. The effects of CX 3 CR1 on macrophage recruitment were assessed using mice of four genotypes: Pou4f3 DTR/+ and either heterozygous or homozygous for CX 3 CR1 ( Pou4f3 DTR/+ :CX 3 CR1 +/− and Pou4f3 DTR/+ :CX 3 CR1 −/− ), and Pou4f3 +/+ and either heterozygous or homozygous for CX 3 CR1 ( Pou4f3 +/+ :CX 3 CR1 +/− and Pou4f3 +/+ :CX 3 CR1 −/− ). Cochlear whole mounts and mid-modiolar sections were processed to image macrophages (CX3CR1 +/GFP , green) and neurons (neurofilament, red). We observed a significant reduction in the numbers of macrophages near the sensory region of the cochlea ( B′ ) and in the spiral ganglion ( E′ ) in CX 3 CR1 −/− mice, compared with CX 3 CR1 +/− mice ( A′ , D′ ) at both 14 d (top) and 56 d (bottom) after DT injection. C , F , G , Quantitative data on macrophages in sensory epithelium at 14 dpi ( C ) and in the spiral ganglia at 56 dpi ( F ). Quantification of macrophages within the spiral ganglia in the middle region of the cochlea at all time points after DT-induced hair cell lesion ( G ). Data are mean ± SD; n = 3–7 for 7 dpi per group; n = 6–7 for 14 dpi per group; n = 3–5 for 28 dpi per group; and n = 9–11 for 56 dpi per group. dpi, days post DT injection; NF, Neurofilament. Statistical significance was computed using two-way ANOVA followed by Tukey's multiple comparison tests. * p

    Journal: The Journal of Neuroscience

    Article Title: Fractalkine Signaling Regulates Macrophage Recruitment into the Cochlea and Promotes the Survival of Spiral Ganglion Neurons after Selective Hair Cell Lesion

    doi: 10.1523/JNEUROSCI.2325-15.2015

    Figure Lengend Snippet: Genetic deletion of CX 3 CR1 led to a reduction in macrophages near the sensory epithelium after hair cell ablation. The effects of CX 3 CR1 on macrophage recruitment were assessed using mice of four genotypes: Pou4f3 DTR/+ and either heterozygous or homozygous for CX 3 CR1 ( Pou4f3 DTR/+ :CX 3 CR1 +/− and Pou4f3 DTR/+ :CX 3 CR1 −/− ), and Pou4f3 +/+ and either heterozygous or homozygous for CX 3 CR1 ( Pou4f3 +/+ :CX 3 CR1 +/− and Pou4f3 +/+ :CX 3 CR1 −/− ). Cochlear whole mounts and mid-modiolar sections were processed to image macrophages (CX3CR1 +/GFP , green) and neurons (neurofilament, red). We observed a significant reduction in the numbers of macrophages near the sensory region of the cochlea ( B′ ) and in the spiral ganglion ( E′ ) in CX 3 CR1 −/− mice, compared with CX 3 CR1 +/− mice ( A′ , D′ ) at both 14 d (top) and 56 d (bottom) after DT injection. C , F , G , Quantitative data on macrophages in sensory epithelium at 14 dpi ( C ) and in the spiral ganglia at 56 dpi ( F ). Quantification of macrophages within the spiral ganglia in the middle region of the cochlea at all time points after DT-induced hair cell lesion ( G ). Data are mean ± SD; n = 3–7 for 7 dpi per group; n = 6–7 for 14 dpi per group; n = 3–5 for 28 dpi per group; and n = 9–11 for 56 dpi per group. dpi, days post DT injection; NF, Neurofilament. Statistical significance was computed using two-way ANOVA followed by Tukey's multiple comparison tests. * p

    Article Snippet: Neuronal peripheral processes and ganglion cell bodies were labeled using a combination of anti-Neurofilament (mouse monoclonal, clone #2H3, Developmental Studies Hybridoma Bank, University of Iowa, 1:100) and anti-β-III tubulin antibodies (mouse monoclonal, catalog #MMS-435P, Covance, 1:500).

    Techniques: Mouse Assay, Injection

    NF-κB regulates mossy fiber projections to CA3 and replenishment of the dentate gyrus. ( a ) Immunostaining for neurofilament M (NF-M) in control mice (IκB/-) reveals a fasciculated organisation of mossy fibers, connecting granule cells to their target cells in CA3. ( b ) Impairment of mossy fiber projections after neuronal NF-κB ablation (IκB/tTA). ( c ) Statistical analysis of NF-M staining shows significantly reduced mossy fiber bundles in IκB/tTA mice ( n  = 8) as compared to controls ( n  = 4) ( P  = 0.0034); ( d ) Synapses stained for synaptophysin are organised in a laminated fashion. Strongest staining was visible in the stratum lucidum (SL), which corresponds to the termination field of mossy fibers on dendrites of CA3 neurons. ( e ) In contrast, staining is strongly reduced in the SL after NF-κB ablation, suggesting an important role of NF-κB in regulating synaptic density. ( f ) NF-κB ablation results also in a significantly decreased dentate gyrus size (white bar in  a ,  b ,  n  = 9) compared to control mice ( n  = 6) ( P  = 

    Journal: PLoS ONE

    Article Title: Regrowing the Adult Brain: NF-?B Controls Functional Circuit Formation and Tissue Homeostasis in the Dentate Gyrus

    doi: 10.1371/journal.pone.0030838

    Figure Lengend Snippet: NF-κB regulates mossy fiber projections to CA3 and replenishment of the dentate gyrus. ( a ) Immunostaining for neurofilament M (NF-M) in control mice (IκB/-) reveals a fasciculated organisation of mossy fibers, connecting granule cells to their target cells in CA3. ( b ) Impairment of mossy fiber projections after neuronal NF-κB ablation (IκB/tTA). ( c ) Statistical analysis of NF-M staining shows significantly reduced mossy fiber bundles in IκB/tTA mice ( n  = 8) as compared to controls ( n  = 4) ( P  = 0.0034); ( d ) Synapses stained for synaptophysin are organised in a laminated fashion. Strongest staining was visible in the stratum lucidum (SL), which corresponds to the termination field of mossy fibers on dendrites of CA3 neurons. ( e ) In contrast, staining is strongly reduced in the SL after NF-κB ablation, suggesting an important role of NF-κB in regulating synaptic density. ( f ) NF-κB ablation results also in a significantly decreased dentate gyrus size (white bar in a , b , n  = 9) compared to control mice ( n  = 6) ( P  = 

    Article Snippet: Antibodies used were: anti-Neurofilament-M (2H3, Developmental Studies Hybridoma Bank), anti-active Caspase-3 (#9664; Cell Signaling), anti-PKA substrate (RRXS/T, #9624; Cell Signaling).

    Techniques: Immunostaining, Mouse Assay, Staining

    Neurite guidance in the 60, 90, 120 μm ACMFP and control (flat PLGA membrane) groups. ( a ) Confocal images show labeling of DAPI (Cyan), Neurofilament (NF-L; Magenta, marker for neurons and neurites) in the 60, 90, 120 μm ACMFPs and control groups (confocal with DIC microscopy). ( b ). Scale bar: 100 μm.

    Journal: Scientific Reports

    Article Title: Aligned contiguous microfiber platform enhances neural differentiation of embryonic stem cells

    doi: 10.1038/s41598-018-24522-9

    Figure Lengend Snippet: Neurite guidance in the 60, 90, 120 μm ACMFP and control (flat PLGA membrane) groups. ( a ) Confocal images show labeling of DAPI (Cyan), Neurofilament (NF-L; Magenta, marker for neurons and neurites) in the 60, 90, 120 μm ACMFPs and control groups (confocal with DIC microscopy). ( b ). Scale bar: 100 μm.

    Article Snippet: Anti-neurofilament (1:200; Santa Cruz Biotechnology) with Dylight 649 conjugated donkey anti-mouse secondary antibodies were used to define neurites outgrowth.

    Techniques: Labeling, Marker, Microscopy

    Expression of Olig1 in sciatic nerves and cultured Schwann cells Immunofluorescent stainings of transversal (A) and longitudinal (C) tissue sections of sciatic nerves from P7 mice revealed Olig1 immunofluorescence in Remak bundles (arrows), identified by a bundle of unmyelinated (MBP-negative) small diameter axons, and in a subset of myelinating Schwann cells (arrowheads). (B) Remak bundle localization was confirmed by the expression of p75 NTR . (D) A progressive increase of Olig1 mRNA expression levels was detected during peripheral nerve development by qRT–PCR. Data were normalized to the expression of 60s, and values at P0 were set to 1. Each data point represent the mean value of at least eight experimental samples, and the error bars indicate the S.D. (E) In vitro , Olig1 expression was predominantly detected in the cytoplasm of cultured Schwann cells (E, arrows), whereas nuclear staining was significantly weaker (E, arrowheads). NF: neurofilament. Bar: A: 100 μm; B, C, E: 20 μm.

    Journal: ASN NEURO

    Article Title: Transcriptional regulation induced by cAMP elevation in mouse Schwann cells

    doi: 10.1042/AN20130031

    Figure Lengend Snippet: Expression of Olig1 in sciatic nerves and cultured Schwann cells Immunofluorescent stainings of transversal (A) and longitudinal (C) tissue sections of sciatic nerves from P7 mice revealed Olig1 immunofluorescence in Remak bundles (arrows), identified by a bundle of unmyelinated (MBP-negative) small diameter axons, and in a subset of myelinating Schwann cells (arrowheads). (B) Remak bundle localization was confirmed by the expression of p75 NTR . (D) A progressive increase of Olig1 mRNA expression levels was detected during peripheral nerve development by qRT–PCR. Data were normalized to the expression of 60s, and values at P0 were set to 1. Each data point represent the mean value of at least eight experimental samples, and the error bars indicate the S.D. (E) In vitro , Olig1 expression was predominantly detected in the cytoplasm of cultured Schwann cells (E, arrows), whereas nuclear staining was significantly weaker (E, arrowheads). NF: neurofilament. Bar: A: 100 μm; B, C, E: 20 μm.

    Article Snippet: Antibodies The following primary antibodies were used: anti-MBP (rat, 1:800, Chemicon), anti-neurofilament (mouse, 1:800, SMI31, Covance), anti-Olig1 (rabbit, 1:1000, Abcam), anti-p75NTR (rabbit, 1:500, Promega), anti-S100 (rabbit, 1:500, Dako).

    Techniques: Expressing, Cell Culture, Mouse Assay, Immunofluorescence, Quantitative RT-PCR, In Vitro, Staining

    Micrographs of SGNs. (A) Bipolar neurons present in cultured SGNs (arrows; magnification, ×100); (B) SGNs stained with anti-neurofilament protein, which is used for the identification of SGNs; green signals can be observed in the membrane (arrow; magnification ×200) rather than in the nuclei. SGNs, spiral ganglion neurons.

    Journal: International Journal of Molecular Medicine

    Article Title: Protective effect of adenovirus-mediated erythropoietin expression on the spiral ganglion neurons in the rat inner ear

    doi: 10.3892/ijmm.2018.3455

    Figure Lengend Snippet: Micrographs of SGNs. (A) Bipolar neurons present in cultured SGNs (arrows; magnification, ×100); (B) SGNs stained with anti-neurofilament protein, which is used for the identification of SGNs; green signals can be observed in the membrane (arrow; magnification ×200) rather than in the nuclei. SGNs, spiral ganglion neurons.

    Article Snippet: Immunohistochemical analysis The anti-neurofilament antibody (1:50; cat. no. ab129349) was obtained from Abcam (Cambridge, UK).

    Techniques: Cell Culture, Staining

    Representative histological features of brainstem GGs from patients 1-4 with representative immunohistochemistry for NF (neurofilament), synaptophysin (SYP), chromogranin (CHR), and VE1 (BRAF), showing large numbers of dysmorphic, irregularly shaped and

    Journal: Brain pathology (Zurich, Switzerland)

    Article Title: Pediatric Brainstem Gangliogliomas Show BRAFV600E Mutation in a High Percentage of Cases

    doi: 10.1111/bpa.12103

    Figure Lengend Snippet: Representative histological features of brainstem GGs from patients 1-4 with representative immunohistochemistry for NF (neurofilament), synaptophysin (SYP), chromogranin (CHR), and VE1 (BRAF), showing large numbers of dysmorphic, irregularly shaped and

    Article Snippet: All cases from our pediatric hospital had originally at the time of diagnosis undergone extensive immunohistochemical characterization utilizing synaptophysin (Biogenex, catalog number Am363; clone Snp88) and either NeuN (Millipore, catalog number MAB377) or neuron-specific enolase(or both, depending on the year the case had originally been diagnosed), plus anti- neurofilament (Cell Marque, clone 2F11) (patients 2, 3, 4, 5, 7, 9, 12) and/or chromogranin (Ventana, catalog number 760-2519) (patients 3, 4, 7, 9).

    Techniques: Immunohistochemistry

    Representative histological features of brainstem GGs from patients 6, 7, 9, 12 with BRAF, NeuN, and neurofilament (NF) immunohistochemistry, showing classic features of GGs, including non-neoplastic lymphocytes (Figures b, c, d, h). Figures a, c, d 200×;

    Journal: Brain pathology (Zurich, Switzerland)

    Article Title: Pediatric Brainstem Gangliogliomas Show BRAFV600E Mutation in a High Percentage of Cases

    doi: 10.1111/bpa.12103

    Figure Lengend Snippet: Representative histological features of brainstem GGs from patients 6, 7, 9, 12 with BRAF, NeuN, and neurofilament (NF) immunohistochemistry, showing classic features of GGs, including non-neoplastic lymphocytes (Figures b, c, d, h). Figures a, c, d 200×;

    Article Snippet: All cases from our pediatric hospital had originally at the time of diagnosis undergone extensive immunohistochemical characterization utilizing synaptophysin (Biogenex, catalog number Am363; clone Snp88) and either NeuN (Millipore, catalog number MAB377) or neuron-specific enolase(or both, depending on the year the case had originally been diagnosed), plus anti- neurofilament (Cell Marque, clone 2F11) (patients 2, 3, 4, 5, 7, 9, 12) and/or chromogranin (Ventana, catalog number 760-2519) (patients 3, 4, 7, 9).

    Techniques: Immunohistochemistry

    Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against neurofilament ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.

    Journal: The Journal of Neuroscience

    Article Title: Patterns of Nogo mRNA and Protein Expression in the Developing and Adult Rat and After CNS Lesions

    doi: 10.1523/JNEUROSCI.22-09-03553.2002

    Figure Lengend Snippet: Localization of Nogo-A in CNS white matter by confocal microscopic analysis. A , No colocalization of Nogo-A (shown by AS 472 in green ) was found with MBP ( red ), which is a major constituent of compact myelin. Nogo-A was expressed in oligodendrocyte cell bodies, their processes, and the inner loop ( B, C , arrowheads ) and outer loop ( B, C , arrows ) of the myelin sheath. The fixation protocol required to demonstrate MBP expression lowered the Nogo-A signal in the inner loop of the myelin membrane, whereas the different protocol used for MAG and MOG immunohistochemistry showed strong expression of Nogo-A in the inner and outer loops of the myelin sheath. E , Nogo-A (AS 472, green ) is expressed in the outer loop ( arrows ) of the myelin sheath and was found to colocalize there with MOG ( red ). G , A strong Nogo-A signal (AS 472, green ) was also found in the inner loop ( arrowheads ), where MAG ( H , red ) is expressed. C, D , Some Nogo-A (AS 472, green ) was also present in axons, as was demonstrated by colocalization ( yellow ) of AS 472 with an antibody against neurofilament ( red ). F , Nogo-A (AS 472, green ) was not present in astrocyte processes stained by an antibody against GFAP ( red ). Scale bar: A, C , 85 μm; B, D–I , 45 μm.

    Article Snippet: For immunohistochemistry the antibodies were diluted in PBS, pH 7.4, containing 1% normal goat serum as follows: mAb IN-1 hybridoma supernatant, 1:5; AS Bruna, 1:7500; AS Bianca, 1:2000; AS 472, 1:2000; AS 818, 1:2000; affinity-purified AS 818, 1:50; anti-myelin-associated glycoprotein (MAG), 1:25 (Roche); anti-myelin oligodendrocyte glycoprotein (MOG), 1:25 (Roche); anti-neurofilament, 1:100 (Roche); and anti-myelin basic protein (MBP), 1:250 (Roche).

    Techniques: Expressing, Immunohistochemistry, Staining

    Degenerative processes in APP 23 mice. ( A and B ) Staining for acetylcholinesterase in 12-month-old transgenic and control mice, respectively. A local distortion of cholinergic fibers in the plaque vicinity ( A ) compared with normal animals ( B ) can be noted. Neuritic spheroids (arrows) are stained with the neurofilament antibody NF200 ( C ). The loss of pyramidal neurons in the vicinity of Aβ deposits in area CA3 is shown in D by toluidine blue staining. (Bars = 50 μm.)

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

    Article Title: Two amyloid precursor protein transgenic mouse models with Alzheimer disease-like pathology

    doi:

    Figure Lengend Snippet: Degenerative processes in APP 23 mice. ( A and B ) Staining for acetylcholinesterase in 12-month-old transgenic and control mice, respectively. A local distortion of cholinergic fibers in the plaque vicinity ( A ) compared with normal animals ( B ) can be noted. Neuritic spheroids (arrows) are stained with the neurofilament antibody NF200 ( C ). The loss of pyramidal neurons in the vicinity of Aβ deposits in area CA3 is shown in D by toluidine blue staining. (Bars = 50 μm.)

    Article Snippet: In addition, anti-heparan sulfate monoclonal antibody 10E4 (Seikagu, Tokyo); anti-neurofilament NF200 (NovoCastra, Newcastle, U.K.); anti-APP 474 raised against purified rat APPs ; anti-apolipoprotein E antibody; anti-MAC-1 and anti-MHC class II (Serotec); anti-GFAP and anti-phosphotyrosine PT-66 (Sigma); anti-mouse complement C3 (Cappel, ICN); anti-PHF1 ( ) recognizing tau phosphoserine 396 and 402; AT8 monoclonal antibody (Innogenetics) ( , ) recognizing phosphoserine 202; R27 ( ) raised against phosphorylated serine 396; R32 ( ) raised against phosphorylated serine 262, N-tau 5 raised against a tau peptide as described , and monoclonal antibody Alz50 ( ) were used.

    Techniques: Mouse Assay, Staining, Transgenic Assay