Journal: JCI Insight

Article Title: Shock waves promote spinal cord repair via TLR3

doi: 10.1172/jci.insight.134552

Figure Lengend Snippet: (A) To investigate possible involvement of NPCs, we quantified nestin-positive cells in spinal cord sections obtained from SWT-treated WT mice. Scale bar: 100 μm. (B) We found a significant increase of nestin-positive cells after treatment, suggesting involvement of NPCs in SWT-induced spinal regeneration. ****P < 0.0001. n = 9 (Sham), n = 9 (CTR), n = 9 (SWT). (C and D) NPCs were isolated from WT and Tlr3–/– mice. We found abundant expression of Tlr3 and nestin in NPCs. Treatment of NPCs with SWT or poly(I:C) increased Tlr3 expression. Scale bar: 20 μm. (E) SWT and poly(I:C) treatment did not affect NPC proliferation, neither in NPCs derived from WT mice nor in those from Tlr3-knockout mice. NPCs obtained from Tlr3-deficient mice showed significantly increased proliferation rates compared with WT NPCs. ****P < 0.0001. n = 6–7. (F) NPCs were treated with SWT or poly(I:C), respectively, and analyzed for the expression of the neuronal cell marker β III tubulin subsequently. Scale bar: 20 μm. (G) Both SWT and poly(I:C) exposure significantly enhanced NPC differentiation in WT but not Tlr3–/– cells, increasing the number of cells expressing β III tubulin. ***P < 0.001, ****P < 0.0001. n = 5. One-way ANOVA with Tukey’s post hoc analysis (B, E, and G). CTR, control; SWT, shock wave therapy; HPF, high-powered field.

Article Snippet: Subsequently, cells were labeled with monoclonal mouse anti–β III tubulin antibody (R&D Systems) for identification of neurons as described previously ( 45 ).

Techniques: Isolation, Expressing, Derivative Assay, Knock-Out, Marker

Journal: Acta physiologica (Oxford, England)

Article Title: Serotonergic paraneurones in the female mouse urethral epithelium and their potential role in peripheral sensory information processing

doi: 10.1111/apha.12919

Figure Lengend Snippet: Primary antibodies used in this study

Article Snippet: Tuj 1 (Neurone-specific class III beta-tubulin) 84 , Neurones , Chicken, affinity purified, Neuromics CH23005 , AB_2210684 , 1 : 1000.

Techniques: Purification, Concentration Assay, Marker, Affinity Purification

Journal: The Journal of cell biology

Article Title: Sirtuin3 ensures the metabolic plasticity of neurotransmission during glucose deprivation.

doi: 10.1083/jcb.202305048

Figure Lengend Snippet: Figure 3. Sirt3 is expressed in neuronal mitochondria and is present in presynaptic terminals. (A) Control hippocampal neurons and neurons transduced with lentivirus expressing Sirt3 shRNA (Sirt3 KD) were immunostained with anti-Sirt3 antibody and the Hoechst nuclear stain. (B) Quantification of Sirt3 immunofluorescence in control and Sirt3 KD neurons. Normalized mean Sirt3 intensity per coverslip ± SEM: Control, 1.02 ± 0.19; Sirt3 KD, 0.34 ± 0.04. n = 4–5 coverslips. (C) Co-immunostaining of neurons with antibodies against Sirt3 and the mitochondrial marker TOMM20. (D) Quantification of Sirt3 colocalization with TOMM20 or Hoechst in neuronal cell bodies. Mean Pearson’s correlation coefficient (r) ± SEM: Sirt3/TOMM20, r = 0.55 ± 0.01, Sirt3/Hoechst: −0.09 ± 0.02. n = 78 cell bodies. (E–G) Coimmunostaining of neurons with antibodies against the neurite marker Tuj1 (E) or presynaptic marker vGLUT1 (F and G). (G) Magnification of the boxed area in F. Arrowheads indicate colocalization of Sirt3 with presynaptic terminals. (H) Quantification of Sirt3 colocalization with vGLUT1 or Hoechst across fields of view (FOVs). Mean Pearson’s correlation coefficient ± SEM: Sirt3/vGLUT1: r = 0.60 ± 0.02, Sirt3/Hoechst: −0.01 ± 0.02. n = 27 FOVs. Scale bars, 10 μm. Mann–Whitney U test (B, D, and H). See Table 1.

Article Snippet: Hippocampal neurons were fixed with 4% PFA, permeabilized with 0.5% Triton, blocked with 5% bovine serum albumin (BSA) for 1 h at room temperature (RT), and incubated at room temperature for 2 h or overnight at 4°C with the following primary antibodies: Sirt3 (1:200, 5490S; Cell Signaling Technology, rabbit, reactivity for human, mouse, and rat), TOMM20 (1:500, ab56783; abcam, mouse, reactivity for human, mouse, and rat), vGLUT1 (1:500, AB5905; Sigma-Aldrich, guinea pig, reactivity for rat), Tuj1 (1:1,000, MAB1195; R&D Systems, mouse, reactivity Table 2.

Techniques: Control, Transduction, Expressing, shRNA, Staining, Immunofluorescence, Immunostaining, Marker, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Glucagon-like peptide 1 receptor (GLP-1R) expression by nerve fibres in inflammatory bowel disease and functional effects in cultured neurons

doi: 10.1371/journal.pone.0198024

Figure Lengend Snippet: Double staining of GLP-1R (Abcam GLP-1R antibody at a dilution of 1:1000, black) with the neuronal marker β Tubulin (at a dilution of 1:1000, red) in, A, sub-mucous plexus of colon from a patient with IBD and, B, human DRG; magnification x20.

Article Snippet: For tissue immunohistochemistry, GLP-1R antibody ab39072 (Abcam, Cambridge, UK) was used at a final titre of 1:1000, CGRP antibody (Merck Millipore AB59200, Watford, UK) was used at a final titre of 1:4000 and rabbit polyclonal anti-human PGP9.5 antibody (RA95101, UltraClone Ltd, UK), was used at 1:40,000, and neuron-specific mouse anti-β-II tubulin/Tuj1 (R&D Systems, Inc, Oxford, UK) at 1:100.

Techniques: Double Staining, Marker

Journal: PLoS ONE

Article Title: Glucagon-like peptide 1 receptor (GLP-1R) expression by nerve fibres in inflammatory bowel disease and functional effects in cultured neurons

doi: 10.1371/journal.pone.0198024

Figure Lengend Snippet: Cultured adult rat DRG neurons (A, B, C) showing β tubulin immunofluorescence (green in A), GLP-1R expression (red in B), and the merged image (C). Bar = 200 μm. High power image of cultured human DRG neurons positive for β tubulin (green in D), intense GLP-1R expression (red) in a small neuron (E), and merged image (F). Bar = 20 μm.

Article Snippet: For tissue immunohistochemistry, GLP-1R antibody ab39072 (Abcam, Cambridge, UK) was used at a final titre of 1:1000, CGRP antibody (Merck Millipore AB59200, Watford, UK) was used at a final titre of 1:4000 and rabbit polyclonal anti-human PGP9.5 antibody (RA95101, UltraClone Ltd, UK), was used at 1:40,000, and neuron-specific mouse anti-β-II tubulin/Tuj1 (R&D Systems, Inc, Oxford, UK) at 1:100.

Techniques: Cell Culture, Immunofluorescence, Expressing

Journal: PLoS ONE

Article Title: Glucagon-like peptide 1 receptor (GLP-1R) expression by nerve fibres in inflammatory bowel disease and functional effects in cultured neurons

doi: 10.1371/journal.pone.0198024

Figure Lengend Snippet: Representative images showing β tubulin immunofluorescence in rat DRG neurons cultured with vehicle (A, control), with Oxm (B), exendin-4 (C), or GLP-1 (D). Bar = 100 microns. Graph showing significantly increased neurite lengths after 48 h treatment with oxyntomodulin (*p = 0.04), exendin-4 (*p = 0.01) and GLP-1 (*p = 0.04) using Student’s unpaired t -test (E). Graph showing lack of significant effect of acute treatment with incretin hormones on capsaicin sensitivity in adult rat DRG neurons (F).

Article Snippet: For tissue immunohistochemistry, GLP-1R antibody ab39072 (Abcam, Cambridge, UK) was used at a final titre of 1:1000, CGRP antibody (Merck Millipore AB59200, Watford, UK) was used at a final titre of 1:4000 and rabbit polyclonal anti-human PGP9.5 antibody (RA95101, UltraClone Ltd, UK), was used at 1:40,000, and neuron-specific mouse anti-β-II tubulin/Tuj1 (R&D Systems, Inc, Oxford, UK) at 1:100.

Techniques: Immunofluorescence, Cell Culture