Journal: Molecular Neurobiology

Article Title: Spared Nerve Injury Causes Sexually Dimorphic Mechanical Allodynia and Differential Gene Expression in Spinal Cords and Dorsal Root Ganglia in Rats

doi: 10.1007/s12035-021-02447-1

Figure Lengend Snippet: L4 dorsal root ganglion (DRG) immunostaining for IB-4 (a) and CGRP (b) after SNI in male (M) and female (F) rats and representative images of staining in L4 (c). **** p < 0.001, ** p < 0.01 by Student’s t -test with Holm-Sidak correction for multiple comparisons and ANOVA. ## p < 0.01 by ANOVA. n = 10

Article Snippet: DRG sections were probed with antibodies for CGRP (1:10,000, Cat# T-4032, Peninsula Laboratories, San Carlos, CA, USA) or FITC-conjugated IB-4 (1:200; Cat# FL-1201, Vector Labs, CA, USA).

Techniques: Immunostaining, Staining

Journal: Frontiers in Neurology

Article Title: Attenuation of immobilization stress-induced hypertension by temperature-controllable warm needle acupuncture in rats and the peripheral neural mechanisms

doi: 10.3389/fneur.2023.1168012

Figure Lengend Snippet: Immunohistochemistry for TRPV1 and/or CGRP, NF200 and IB4 in dorsal root ganglia (DRGs) following warm needle acupuncture instrument (WAI) stimulation at PC-6. (A,B) Fluorescence intensity and representative images of TRPV1 and CGRP double-stained DRG neurons (yellow) in the normal ( n = 6) and WAI ( n = 6) groups. *** indicates a significance level of p < 0.001 vs. Normal. Immunofluorescent images for TRPV1 (red) and CGRP (green) (C,D) Fluorescence intensity and representative images of TRPV1- and NF200-positive DRG neurons in the normal ( n = 6) and WAI ( n = 6) groups. Immunofluorescent images for TRPV1 (red) and NF200 (green) (E,F) Fluorescence intensity and representative images of TRPV1- and IB4-positive DRG neurons in the normal ( n = 6) and WAI ( n = 6) groups. * p < 0.05 vs. Normal. Double-stained cells (yellow) by TRPV1 (green)/IB4 (red). Scale bar: 100 μm.

Article Snippet: The slides were incubated with mouse anti-TRPV1 antibody (1:500, Santa Cruz Biotechnology), rabbit anti-calcitonin gene-related peptide (CGRP, 1:1000, Abcam), anti-lectin IB4 Alexa Fluor 488 conjugated to FITC (1:500, Invitrogen), rabbit anti-neurofilament 200 (NF200, 1:1000, Sigma–Aldrich) and mouse monoclonal PGP 9.5 antibody (1:1000; Abcam, Cambridge, United Kingdom; ab8189) overnight at 4°C, followed by incubation with secondary antibodies (1:200, Alexa Fluor 488 donkey anti-rabbit IgG antibody), Thermo Scientific, MA, United States, Product# A-21206; 1:200, Alexa Fluor ® 594 donkey anti-mouse IgG antibody, Thermo Scientific, Product# A-21203, donkey anti-mouse IgG Cy3 (1:500, 133,644, Invitrogen).

Techniques: Immunohistochemistry, Fluorescence, Staining

Journal: The Journal of Neuroscience

Article Title: DRG Voltage-Gated Sodium Channel 1.7 Is Upregulated in Paclitaxel-Induced Neuropathy in Rats and in Humans with Neuropathic Pain

doi: 10.1523/JNEUROSCI.0899-17.2017

Figure Lengend Snippet: The expression of Nav1.7 is increased in nociceptive DRG neurons in rats with paclitaxel-treated CIPN. Representative IHC images are shown in A–H. A, B, The expression of Nav1.7 (red) in DRGs is normally low in naive (data not shown) and vehicle-treated rats (A) but becomes greatly increased by day 7 after paclitaxel treatment (B). C–H, Nav1.7 is colocalized in subsets of IB4-positive (green) neurons (C, D, colocalization indicated in yellow) and CGRP-positive (blue) neurons (E, F, colocalization indicated in purple), as well as in a percentage of neurons negative for both IB4 and CGRP (G, H, overall merged). I–K, The bar graphs summarize the grouped IHC data and show that the overall number of Nav1.7-positive neurons (I) was significantly higher in the paclitaxel-treated rats (black bars) than in the vehicle-treated rats (white bars); and these increases were significantly higher in neurons with diameters <30 μm and to a lesser extent in medium-sized (30–45 μm) DRG neurons (J); and in IB4-positive and CGRP-positive neurons, as well as neurons negative for both markers (K). Scale bar, 100 μm. *p < 0.05, ***p < 0.001.

Article Snippet: After blocking in 5% normal donkey serum and 0.2% Triton X-100 in PBS for 1 h at room temperature, the sections were incubated overnight at 4°C in 1% normal donkey serum and 0.2% Triton X-100 in PBS containing primary antibodies against the following targets: Na v 1.7 (rabbit anti-rat, 1:500; catalog #ASC-008, Alomone Labs); IB4 (1:1000; BS-Isolectin B4 FITC Conjugate, Sigma-Aldrich); calcitonin gene-related peptide (CGRP; mouse anti-rat, 1:1000; Abcam); NeuN (mouse anti-rat, 1:1000; Millipore); glial fibrillary acid protein (GFAP; mouse anti-rat, 1:1000; Cell Signaling Technology); and OX-42 (mouse anti-rat, 1:1000; Serotec).

Techniques: Expressing

Journal: The Journal of Neuroscience

Article Title: DRG Voltage-Gated Sodium Channel 1.7 Is Upregulated in Paclitaxel-Induced Neuropathy in Rats and in Humans with Neuropathic Pain

doi: 10.1523/JNEUROSCI.0899-17.2017

Figure Lengend Snippet: The expression of Nav1.7 is increased and colocalized with IB4, CGRP, and NeuN in spinal cord segments L4–L5 at day 7 after treatment with paclitaxel. A, B, Representative immunohistochemical (IHC) images show that expression of Nav1.7 (red) in spinal cord was quite pronounced compared with vehicle (A) by day 7 after treatment with paclitaxel (B). In C, the absorption control shows no staining. D, E, Double IHC shows that Nav1.7 (red) was expressed in some IB4-positive (D, green, with colocalization shown in yellow) and especially were expressed in CGRP-positive (blue) neuron terminals (E, colocalization indicated in purple). In F, the merged image indicates that Nav1.7 expression was upregulated in a large percentage of peripheral afferent terminals positive for CGRP. G–I show that Nav1.7 was expressed in NeuN-positive spinal cells in deeper lamina following paclitaxel treatment. J–L are a further test of antibody specificity as no expression of Nav1.7 was found in the cerebellum. Scale bars: lower-scale images, 200 μm; higher-magnification images, 50 μm.

Article Snippet: After blocking in 5% normal donkey serum and 0.2% Triton X-100 in PBS for 1 h at room temperature, the sections were incubated overnight at 4°C in 1% normal donkey serum and 0.2% Triton X-100 in PBS containing primary antibodies against the following targets: Na v 1.7 (rabbit anti-rat, 1:500; catalog #ASC-008, Alomone Labs); IB4 (1:1000; BS-Isolectin B4 FITC Conjugate, Sigma-Aldrich); calcitonin gene-related peptide (CGRP; mouse anti-rat, 1:1000; Abcam); NeuN (mouse anti-rat, 1:1000; Millipore); glial fibrillary acid protein (GFAP; mouse anti-rat, 1:1000; Cell Signaling Technology); and OX-42 (mouse anti-rat, 1:1000; Serotec).

Techniques: Expressing, Immunohistochemical staining, Staining