Journal: PLoS ONE

Article Title: Immunohistochemical, Ultrastructural and Functional Analysis of Axonal Regeneration through Peripheral Nerve Grafts Containing Schwann Cells Expressing BDNF, CNTF or NT3

doi: 10.1371/journal.pone.0069987

Figure Lengend Snippet: (D) Quantification revealed significantly greater numbers of IB 4 + axons in normal nerves compared to acellular grafts (*) and in NT3 grafts compared to all other experimental groups (**). (E) The number of CGRP + axons was not significantly different between experimental groups. Values represent M ± SEM of n = 3; p<0.05. Further details on statistical analysis provided as . Scale bar for A–C: 100 µm.

Article Snippet: Immunostaining was done at room temperature for antibodies to calcitonin gene-related peptide (CGRP) (AbDSerotec #1720-9007, 1∶1000) to label axons of primary peptidergic sensory neurons.

Techniques:

Journal: PLoS ONE

Article Title: Immunohistochemical, Ultrastructural and Functional Analysis of Axonal Regeneration through Peripheral Nerve Grafts Containing Schwann Cells Expressing BDNF, CNTF or NT3

doi: 10.1371/journal.pone.0069987

Figure Lengend Snippet: Summary of data.

Article Snippet: Immunostaining was done at room temperature for antibodies to calcitonin gene-related peptide (CGRP) (AbDSerotec #1720-9007, 1∶1000) to label axons of primary peptidergic sensory neurons.

Techniques: Functional Assay

Journal: iScience

Article Title: A novel Na v 1.8-FLPo driver mouse for intersectional genetics to uncover the functional significance of primary sensory neuron diversity

doi: 10.1016/j.isci.2024.109396

Figure Lengend Snippet:

Article Snippet: goat anti-CGRP , OriGene , cat# BP022.

Techniques: Recombinant, Electron Microscopy, Labeling, Plasmid Preparation, Mutagenesis, Cloning, Generated, Software

Journal: Frontiers in Immunology

Article Title: Peripheral and Central Neuroinflammatory Changes and Pain Behaviors in an Animal Model of Multiple Sclerosis

doi: 10.3389/fimmu.2016.00369

Figure Lengend Snippet: Specific damage to myelinated A-class fibers in clinical EAE . Representative images of NF200/ATF-3 double-labeling showing the presence of NF200+ATF-3+ cells in the trigeminal ganglion of EAE mice (B) , but not control mice (A) at day 16. Scale bar equals 25 μm and white arrow indicates an NF200 immunoreactive cell with ATF-3 nuclear staining. (C) Bar graph showing significantly increased numbers of NF200+ATF-3+ cell bodies compared to CGRP+ATF-3+ and IB4+ATF-3+ cell bodies in the trigeminal ganglia of EAE mice. ** P < 0.01 and **** P < 0.0001, one-way ANOVA followed by Tukey’s post hoc test, n = 4–5. Data are expressed as mean ± SEM.

Article Snippet: The blocking solution was drained, and sections were incubated with the following antibodies diluted in PBS containing 5% bovine serum albumin and 0.03% Triton-X: rat anti-mouse CD3 (T cells; 1:100; R&D systems, MN, USA), rabbit anti-mouse/rat IBA-1 (ionized calcium-binding adaptor-1; macrophages/microglia; 1:2000; Wako Chemicals USA, Richmond, VA, USA), mouse anti-mouse GFAP (glial fibrillary acidic protein; activated astrocytes; 1:2000; Chemicon, Temecula, CA, USA), rabbit anti-mouse ATF-3 (activating transcription factor-3; cell damage; 1:400; Santa Cruz Biotechnology Inc., TX, USA), goat anti-mouse CGRP (calcitonin gene-related peptide; peptidergic neurons; 1:1000; Abcam, VIC, Australia), anti-mouse IB4-FITC (isolectin B4; non-peptidergic neurons; 1:100; Sigma Aldrich), and rabbit anti-mouse NF200 (neurofilament 200; myelinated neurons; 1:1000; Sigma Aldrich).

Techniques: Labeling, Control, Staining

Journal: Journal of Diabetes Research

Article Title: Glucagon-Like Peptide-1 Receptor Agonist Protects Dorsal Root Ganglion Neurons against Oxidative Insult

doi: 10.1155/2019/9426014

Figure Lengend Snippet: Distribution of sensory neuronal markers in the dorsal root ganglion (DRG) neuron cell line treated with exendin-4. Pictures on the left side are neurons without any treatment. Pictures on the right side are neurons treated with 100 nM exendin-4 for 36 hours. TRPV1: red (a), substance P: green (b), CGRP: green: DAPI (c), scale 100 μ m.

Article Snippet: The cells were blocked with 1% bovine serum albumin, and the following primary antibodies were applied at 4°C overnight: rabbit polyclonal anti-TRPV1 antibody (1 : 200; Neuromics, Northfield, MN, USA), goat polyclonal anti-substance P antibody (1 : 200; Santa Cruz, Santa Cruz, CA, USA), and goat polyclonal anti-CGRP antibody (1 : 200; Santa Cruz).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Focal Lysolecithin-Induced Demyelination of Peripheral Afferents Results in Neuropathic Pain Behavior That Is Attenuated by Cannabinoids

doi: 10.1523/JNEUROSCI.23-08-03221.2003

Figure Lengend Snippet: Immunohistochemical colocalization of neuronal subtype markers with ATF3, NPY, and the sodium channel subtypes Nav1.8 and Nav1.3 in DRG cells after lysolecithin treatment of the sciatic nerve. a, Expression of ATF3 in ipsilateral and contralateral DRG sections from lysolecithin-treated animals on day 13 after treatment (n = 4). There was no significant increase in expression of ATF3 from that seen in control DRG after lysolecithin treatment or sham treatment. The statistical significance (*p < 0.05) of any difference was determined by a Kruskal–Wallis one-way ANOVA on ranks with all pair-wise multiple comparison procedures (Dunn's method). Images show nuclei of ATF3-immunopositive cells labeled with FITC (green) and NF-200-immunopositive cells labeled with TRITC (red), in ipsilateral (i) and contralateral (ii) DRG from lysolecithin-treated animals and ipsilateral DRG (iii) from CCI animals. Scale bar, 20 μm.b, Expression of the neuropeptide NPY in ipsilateral and contralateral DRG sections from lysolecithin-treated animals at 13 d after treatment (n = 4). Images show NF-200-immunopositive cells labeled with TRITC (red) and NF-200/NPY-colocalized, immunopositive cells labeled with FITC (green) in DRG ipsilateral to lysolecithin treatment (i) as opposed to no positive cells in DRG contralateral to lysolecithin treatment (ii). Scale bar, 20 μm. c, The expression of the Nav1.8 channel in ipsilateral and contralateral DRG sections from lysolecithin-treated animals on day 13 after treatment (n = 4). After lysolecithin treatment of the sciatic nerve, there was a decrease in the number of cells expressing Nav1.8 in ipsilateral DRG compared with contralateral DRG, all of which were immunopositive for NF-200. There was no significant decrease in expression after sham treatment. The statistical significance (*p < 0.05) of any difference was determined by a Kruskal–Wallis one-way ANOVA on ranks with all pair-wise multiple comparison procedures (Dunn's method). Images show Nav1.8-immunopositive cells labeled with FITC (green) and NF-200-immunopositive cells labeled with TRITC (red), with coexpression appearing yellow in ipsilateral (i) and contralateral (ii) DRG from lysolecithin-treated animals. Scale bar, 20 μm. d, Expression of the Nav1.3 channel in ipsilateral and contralateral DRG sections from lysolecithin-treated animals 13 d after treatment and sham-treated animals on the same day after treatment (n = 4 in each case). Images show NF-200-immunopositive cells labeled with TRITC (red) and Nav1.3-immunopositive cells labeled with FITC (green) with coexpression appearing yellow in DRG ipsilateral to lysolecithin treatment (i) as opposed to no positive cells in DRG contralateral to lysolecithin treatment (ii). Scale bar, 20 μm.

Article Snippet: Sections were then washed in buffer and incubated with the appropriate secondary antibodies linked to either tetramethyl rhodamine isothiocyanate (TRITC) (goat anti-mouse-TRITC, 1:200, to detect all mouse primary antisera; Southern Biotechology, Birmingham, AL) or fluorescein isothiocyanate (FITC) (goat anti-rabbit-FITC, 1:200, to detect all goat primary antisera; Cappell; donkey anti-sheep-FITC, 1:100, to detect anti-CGRP primary antiserum; Jackson ImmunoResearch Laboratories, West Grove, PA) for 2 hr at room temperature.

Techniques: Immunohistochemical staining, Expressing, Control, Comparison, Labeling

Journal:

Article Title: Mast cell degranulation activates a pain pathway underlying migraine headache

doi: 10.1016/j.pain.2007.03.012

Figure Lengend Snippet: Localization of pERK in CGRP-immunoreactive dural nerve fibers following mast cell degranulation with 48/80. (A) Dural whole-mount section showing pERK immunofluorescence in two dural nerve fibers (arrows) following treatment with 48/80. The same two fibers (arrowheads) showing CGRP-immunofluorescence are shown in C. The merged image (pERK+CGRP) in E shows that the pERK and CGRP labeling is overlapping and thus localized within the same fibers. The dural whole-mount section shown on the right (B, D, F) was obtained from an animal treated with sodium cromoglycate (SCG) prior to 48/80 and shows lack of pERK-immunofluorescence (B), but preservation of CGRP labeling in 3 dural fibers (arrowheads in D). Panel F represents a merged image of B and D. Scale bar = 50 μm.

Article Snippet: For double immunofluorescence detection, dura specimens were incubated with a mixture of the monoclonal anti-pERK and a polyclonal goat anti-CGRP antibodies (T-5027, 1:500, Bachem, San Carlos, CA ) followed by incubating with a mixture of appropriate Alexa Fluor 488 (Invitrogen, Carlsbad, CA) and Cy3 (Sigma, St. Louis, MO) conjugated secondary antibodies.

Techniques: Immunofluorescence, Labeling, Preserving