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p2x3r apr 026 (Alomone Labs)

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Alomone Labs p2x3r apr 026
P2x3r Apr 026, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/p2x3r+apr+026/Anti-P2X3+Receptor+(extracellular)+Antibody/pm41990747-253-28-30
Average 94 stars, based on 18 article reviews

p2x3r apr 026 - by Bioz Stars, 2026-10

94/100 stars

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Transfection:

Article Title: Skin inflammation and itch response are independently regulated by distinct nociceptor subsets.
Article Snippet: 4μm sections were stained with H&E, imaged using a LEICA DM4000B microscope and analyzed with Fiji. .. They were then Antibody Reference Atf3 NBP1-85816 (Novius) TH AB1542 (Merck) Tubb3 (b-tubulin) 801210 (Biolegend) IB4-488 I21411 (Thermofisher) Trpv1 ACC-030 (Alomone) Substance P MAB356 (Merck) Gfrα2 AF429 (Bio-Techne) P2X3R APR-026 (Alomone) NFH AB1989 (Merck) e4 Immunity 59, 1–16.e1–e9, May 12, 2026 transfected with 2 μg of TRPM8, TRPV1, TRPV4 or TRPA1 plus GCamp6 (0.3μg) using the calcium phosphate method. ..

Image Search Results

Sorted PSN-derived exosomal miR-1306-3p activated spinal P2X3R and enhanced synaptic transmission, thus leading to visceral pain. (A) Schematic diagram and representative images of sorting PSN-derived exosomes from spinal dorsal horn. (B) The expressions of miR-1306-3p, miR-1949, miR-185-5p, and miR-324-5p were detected in the PSN-derived exosomes from spinal dorsal horn of CON and NMD mice by Q-PCR (n = 4 mice for each group, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). (C) Fluorescent representation of co-localization of GFP (green), P2X3R (red), and DAPI (blue) in the spinal dorsal horn. Scale bar = 50 μm. Representative 3D confocal images on the right was the enlarged view in the white box of merge. Scale bar = 20 μm. (D) Representative traces of sEPSCs were recorded in the T13 to L2 spinal dorsal horn neurons before and after the incubation of miR-1306-3p and Gefapixant. Histogram of the amplitude and frequency of sEPSCs in Pre, miR-1306-3p, and miR-1306-3p+Gefapixant group (n = 10 cells from 4 mice for each group, ** P < 0.01, *** P < 0.001, one-way ANOVA followed by Tukey post hoc test). (E) Statistical diagram of the visceral pain threshold in NMD mice after injection of antagomir-miR-1306-3p or antagomir-negative control (antagomir-NC) at Pre, 0.5, 1, 2, and 4 hours (antagomir-NC: n = 6 mice, antagomir-miR-1306-3p: n = 7 mice, ** P < 0.01, 2-way ANOVA followed by Sidak post hoc test). (F) Statistical diagram of the visceral pain threshold in NMD mice after injection of Gefapixant at Pre, 0.5, 1, 2, 4, and 8 hours (n = 7 mice for each group, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA followed by Dunnett post hoc test). (G) Histogram of the visceral pain thresholds in CON mice treated with agomir-negative control (agomir-NC) or agomir-miR-1306-3p at Pre, 0.5, 1, 2, and 4 hours, respectively (n = 6 mice for each group, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). (H) Histogram of the visceral pain thresholds in CON mice treated with agomir-miR-1306-3p+NS or agomir-miR-1306-3p+Gefapixant at Pre, 0.5, 1, 2, and 4 hours, respectively (n = 6 mice for each group, * P < 0.05, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). ANOVA, analysis of variance; NMD, neonatal maternal deprivation; PSN, primary sensory neuron; Q-PCR, real-time quantitative polymerase chain reaction; sEPSC, spontaneous excitatory postsynaptic current.

Journal: Pain

Article Title: Potentiation of visualized exosomal miR-1306-3p from primary sensory neurons contributes to chronic visceral pain via spinal P2X3 receptors

doi: 10.1097/j.pain.0000000000003537

Figure Lengend Snippet: Sorted PSN-derived exosomal miR-1306-3p activated spinal P2X3R and enhanced synaptic transmission, thus leading to visceral pain. (A) Schematic diagram and representative images of sorting PSN-derived exosomes from spinal dorsal horn. (B) The expressions of miR-1306-3p, miR-1949, miR-185-5p, and miR-324-5p were detected in the PSN-derived exosomes from spinal dorsal horn of CON and NMD mice by Q-PCR (n = 4 mice for each group, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). (C) Fluorescent representation of co-localization of GFP (green), P2X3R (red), and DAPI (blue) in the spinal dorsal horn. Scale bar = 50 μm. Representative 3D confocal images on the right was the enlarged view in the white box of merge. Scale bar = 20 μm. (D) Representative traces of sEPSCs were recorded in the T13 to L2 spinal dorsal horn neurons before and after the incubation of miR-1306-3p and Gefapixant. Histogram of the amplitude and frequency of sEPSCs in Pre, miR-1306-3p, and miR-1306-3p+Gefapixant group (n = 10 cells from 4 mice for each group, ** P < 0.01, *** P < 0.001, one-way ANOVA followed by Tukey post hoc test). (E) Statistical diagram of the visceral pain threshold in NMD mice after injection of antagomir-miR-1306-3p or antagomir-negative control (antagomir-NC) at Pre, 0.5, 1, 2, and 4 hours (antagomir-NC: n = 6 mice, antagomir-miR-1306-3p: n = 7 mice, ** P < 0.01, 2-way ANOVA followed by Sidak post hoc test). (F) Statistical diagram of the visceral pain threshold in NMD mice after injection of Gefapixant at Pre, 0.5, 1, 2, 4, and 8 hours (n = 7 mice for each group, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA followed by Dunnett post hoc test). (G) Histogram of the visceral pain thresholds in CON mice treated with agomir-negative control (agomir-NC) or agomir-miR-1306-3p at Pre, 0.5, 1, 2, and 4 hours, respectively (n = 6 mice for each group, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). (H) Histogram of the visceral pain thresholds in CON mice treated with agomir-miR-1306-3p+NS or agomir-miR-1306-3p+Gefapixant at Pre, 0.5, 1, 2, and 4 hours, respectively (n = 6 mice for each group, * P < 0.05, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). ANOVA, analysis of variance; NMD, neonatal maternal deprivation; PSN, primary sensory neuron; Q-PCR, real-time quantitative polymerase chain reaction; sEPSC, spontaneous excitatory postsynaptic current.

Article Snippet: The primary antibodies were anti-Rab27a (1:200, Cat no. 168013; Synaptic Systems, Göttingen, Germany, RRID: AB_887766), anti-GFP-FITC (1:500, Cat no. ab6662; Abcam, Cambridge, United Kingdom, RRID: AB_305635), anti-CD63 (1:100, Cat no. sc-5275; Santa Cruz Biotechnology, Dallas, TX, RRID: AB_627877), anti-NeuN (1:50, Cat no. MAB377; Merck Millipore, Burlington, MA, RRID: AB_2298772), anti-glutamine synthetase (1:500, Cat no. ab64613; Abcam, RRID: AB_1140869), anti-CGRP (calcitonin gene-related peptide) (1:100, Cat no. C7113; Sigma-Aldrich, St. Louis, MO, RRID: AB_259000), anti-IB4 + -FITC (1:200, Cat no. L-1104; Vector Laboratories, Newark, CA, RRID: AB_2336498), anti-NF200 (1:200, Cat no. ab213128; Abcam, RRID: AB_3073795), anti-GFAP (glial fibrillary acidic protein) (1:100, Cat no. 3670; Cell Signaling Technology, RRID: AB_561049), anti-Iba-1 (1:100, Cat no. ab5076; Abcam, RRID: AB_2224402), and P2X3R (1:200, Cat no. APR-026; Alomone Labs, Jerusalem, Israel, RRID: AB_2341052).

Techniques: Derivative Assay, Transmission Assay, Incubation, Injection, Negative Control, Real-time Polymerase Chain Reaction

siR-Rab27a reduced visible PSN-derived exosomes, suppressed spinal synaptic transmission, and alleviated visceral pain. (A and B) Representative images and fluorescence area of GFP were quantified in the left spinal dorsal horn of the NMD mice injected with siR-NC or siR-Rab27a (siR-NC: n = 3 mice, siR-Rab27a: n = 4 mice, * P < 0.05, 2-sample t test). Scale bar = 50 μm. (C) The protein expression of GFP in the left spinal dorsal horn of siR-NC and siR-Rab27a mice (siR-NC: n = 5 mice, siR-Rab27a: n = 4 mice, * P < 0.05, 2 sample t test). (D) Representative images of GFP, NeuN-labeled neurons, and DAPI-labeled nucleus in the left spinal dorsal horn of siR-NC and siR-Rab27a mice. Scale bar = 50 μm. The white box in the lower right corner was the typical GFP + positive neurons, scale bar = 10 μm. (E) Quantification of the percentage of GFP + positive neurons in the spinal dorsal horn neurons of siR-NC and siR-Rab27a mice (siR-NC: n = 4 mice, siR-Rab27a: n = 5 mice, * P < 0.05, Mann–Whitney test). (F) Representative images of GFP, P2X3R-positive neurons and DAPI in the left spinal dorsal horn of siR-NC and siR-Rab27a mice. Scale bar = 50 μm. The white box in the lower right corner was the typical GFP + P2X3R + positive neurons. Scale bar = 10 μm. (G) Quantification of the percentage of GFP + P2X3R + positive neurons in spinal P2X3R-positive neurons of siR-NC and siR-Rab27a mice (n = 4 mice for each group, * P < 0.05, Mann–Whitney test). (H) Representative traces of sEPSCs were recorded in the T13 to L2 spinal dorsal horn neurons of siR-NC and siR-Rab27a mice. Histogram and cumulative probability distributions of the amplitude and frequency of sEPSCs in siR-NC and siR-Rab27a mice (siR-NC: n = 8 cells from 3 mice, siR-Rab27a: n = 6 cells from 3 mice, * P < 0.05, 2-sample t test). (I) The visceral pain threshold at Pre, 1, 2, 3, 4, and 5 days in NMD mice injected with siR-NC or siR-Rab27a, respectively (n = 8 mice for each group, ** P < 0.01, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). (J) Statistical diagram of the time on the rod in the siR-NC or siR-Rab27a group at Pre, 1, 2, 3, 4, and 5 days, respectively (n = 8 mice for each group, P > 0.05, 2-way ANOVA followed by Sidak post hoc test). ANOVA, analysis of variance; GFP, green fluorescent protein; NMD, neonatal maternal deprivation; PSN, primary sensory neuron; sEPSC, spontaneous excitatory postsynaptic current.

Journal: Pain

Article Title: Potentiation of visualized exosomal miR-1306-3p from primary sensory neurons contributes to chronic visceral pain via spinal P2X3 receptors

doi: 10.1097/j.pain.0000000000003537

Figure Lengend Snippet: siR-Rab27a reduced visible PSN-derived exosomes, suppressed spinal synaptic transmission, and alleviated visceral pain. (A and B) Representative images and fluorescence area of GFP were quantified in the left spinal dorsal horn of the NMD mice injected with siR-NC or siR-Rab27a (siR-NC: n = 3 mice, siR-Rab27a: n = 4 mice, * P < 0.05, 2-sample t test). Scale bar = 50 μm. (C) The protein expression of GFP in the left spinal dorsal horn of siR-NC and siR-Rab27a mice (siR-NC: n = 5 mice, siR-Rab27a: n = 4 mice, * P < 0.05, 2 sample t test). (D) Representative images of GFP, NeuN-labeled neurons, and DAPI-labeled nucleus in the left spinal dorsal horn of siR-NC and siR-Rab27a mice. Scale bar = 50 μm. The white box in the lower right corner was the typical GFP + positive neurons, scale bar = 10 μm. (E) Quantification of the percentage of GFP + positive neurons in the spinal dorsal horn neurons of siR-NC and siR-Rab27a mice (siR-NC: n = 4 mice, siR-Rab27a: n = 5 mice, * P < 0.05, Mann–Whitney test). (F) Representative images of GFP, P2X3R-positive neurons and DAPI in the left spinal dorsal horn of siR-NC and siR-Rab27a mice. Scale bar = 50 μm. The white box in the lower right corner was the typical GFP + P2X3R + positive neurons. Scale bar = 10 μm. (G) Quantification of the percentage of GFP + P2X3R + positive neurons in spinal P2X3R-positive neurons of siR-NC and siR-Rab27a mice (n = 4 mice for each group, * P < 0.05, Mann–Whitney test). (H) Representative traces of sEPSCs were recorded in the T13 to L2 spinal dorsal horn neurons of siR-NC and siR-Rab27a mice. Histogram and cumulative probability distributions of the amplitude and frequency of sEPSCs in siR-NC and siR-Rab27a mice (siR-NC: n = 8 cells from 3 mice, siR-Rab27a: n = 6 cells from 3 mice, * P < 0.05, 2-sample t test). (I) The visceral pain threshold at Pre, 1, 2, 3, 4, and 5 days in NMD mice injected with siR-NC or siR-Rab27a, respectively (n = 8 mice for each group, ** P < 0.01, *** P < 0.001, 2-way ANOVA followed by Sidak post hoc test). (J) Statistical diagram of the time on the rod in the siR-NC or siR-Rab27a group at Pre, 1, 2, 3, 4, and 5 days, respectively (n = 8 mice for each group, P > 0.05, 2-way ANOVA followed by Sidak post hoc test). ANOVA, analysis of variance; GFP, green fluorescent protein; NMD, neonatal maternal deprivation; PSN, primary sensory neuron; sEPSC, spontaneous excitatory postsynaptic current.

Techniques: Derivative Assay, Transmission Assay, Fluorescence, Injection, Expressing, Labeling, MANN-WHITNEY

A working model showing that exosomes in the circuit of DRG-spinal cord leading to chronic visceral pain by exosome visualization technologies. The upregulation of Rab27a in colon-related DRGs promoted the release of exosomal miR-1306-3p, which activated P2X3R in the spinal dorsal horn neurons and enhanced synaptic transmission, thereby contributing to chronic visceral pain. Meanwhile, intrathecal injection of GW4869 or siR-Rab27a reduced visible PSN-derived exosomes in spinal cord, suppressed spinal synaptic transmission, and alleviated visceral pain in NMD mice. Inhibition of P2X3R also alleviated visceral pain by intrathecal injection of Gefapixant. DRG, dorsal root ganglia; EPSC, excitatory postsynaptic currents; MVB, multivesicular bodies; NMD, neonatal maternal deprivation; PSN, primary sensory neuron.

Journal: Pain

Article Title: Potentiation of visualized exosomal miR-1306-3p from primary sensory neurons contributes to chronic visceral pain via spinal P2X3 receptors

doi: 10.1097/j.pain.0000000000003537

Figure Lengend Snippet: A working model showing that exosomes in the circuit of DRG-spinal cord leading to chronic visceral pain by exosome visualization technologies. The upregulation of Rab27a in colon-related DRGs promoted the release of exosomal miR-1306-3p, which activated P2X3R in the spinal dorsal horn neurons and enhanced synaptic transmission, thereby contributing to chronic visceral pain. Meanwhile, intrathecal injection of GW4869 or siR-Rab27a reduced visible PSN-derived exosomes in spinal cord, suppressed spinal synaptic transmission, and alleviated visceral pain in NMD mice. Inhibition of P2X3R also alleviated visceral pain by intrathecal injection of Gefapixant. DRG, dorsal root ganglia; EPSC, excitatory postsynaptic currents; MVB, multivesicular bodies; NMD, neonatal maternal deprivation; PSN, primary sensory neuron.

Techniques: Transmission Assay, Injection, Derivative Assay, Inhibition

Inhibiting P2X3R Reduces Naked Mole-Rat Pain Response to Formalin. Inhibition of P2X3r decreases pain responses induced by formalin in naked mole-rats in Phase I (A) and Phase II (B) but has no effect on those responses in mice. P2X3r Ant = Antagonist A-317491. (n = 8 mice and NMR saline control; 4 mice and 5 NMR with antagonist; * p < .05, *** p < .001, Statistical analysis was performed using a paired T-test).

Journal: Neurobiology of Pain

Article Title: The naked mole-rat has a functional purinergic pain pathway despite having a non-functional peptidergic pain pathway

doi: 10.1016/j.ynpai.2020.100047

Figure Lengend Snippet: Inhibiting P2X3R Reduces Naked Mole-Rat Pain Response to Formalin. Inhibition of P2X3r decreases pain responses induced by formalin in naked mole-rats in Phase I (A) and Phase II (B) but has no effect on those responses in mice. P2X3r Ant = Antagonist A-317491. (n = 8 mice and NMR saline control; 4 mice and 5 NMR with antagonist; * p < .05, *** p < .001, Statistical analysis was performed using a paired T-test).

Article Snippet: Primary antibodies used were rabbit anti-P2X3r polyclonal (1:1000, Alomone) and guinea pig anti-TRPV1 polyclonal (1:500, Alomone).

Techniques: Inhibition

P2X3R is Functional in Cultured Naked Mole-Rat Dorsal Root Ganglion Neurons. Functionality of P2X3r was assessed with IHC and Ca 2+ -imaging. Example images of P2X3r and TRPV1 expression in DRG neurons of mice (A, n = 3) and naked mole-rats (C, n = 3). Quantification of neurons by cell area in mice (B) and naked mole-rats (D) shows a similar distribution of P2X3r and TRPV1 expression/co-expression but note the enhanced peak of small diameter fibers in mouse compared to naked mole-rat. Quantification of P2X3r expression in mouse and naked mole-rat DRG reveals no significant difference (E, n = 3). Responsiveness of mouse and naked mole-rat DRG neurons to the P2X3r agonist αβ-meATP is similar across species (F, n = 4). Statistical analysis of data in E and F was performed using Mann Whitney U test.

Journal: Neurobiology of Pain

Article Title: The naked mole-rat has a functional purinergic pain pathway despite having a non-functional peptidergic pain pathway

doi: 10.1016/j.ynpai.2020.100047

Figure Lengend Snippet: P2X3R is Functional in Cultured Naked Mole-Rat Dorsal Root Ganglion Neurons. Functionality of P2X3r was assessed with IHC and Ca 2+ -imaging. Example images of P2X3r and TRPV1 expression in DRG neurons of mice (A, n = 3) and naked mole-rats (C, n = 3). Quantification of neurons by cell area in mice (B) and naked mole-rats (D) shows a similar distribution of P2X3r and TRPV1 expression/co-expression but note the enhanced peak of small diameter fibers in mouse compared to naked mole-rat. Quantification of P2X3r expression in mouse and naked mole-rat DRG reveals no significant difference (E, n = 3). Responsiveness of mouse and naked mole-rat DRG neurons to the P2X3r agonist αβ-meATP is similar across species (F, n = 4). Statistical analysis of data in E and F was performed using Mann Whitney U test.

Article Snippet: Primary antibodies used were rabbit anti-P2X3r polyclonal (1:1000, Alomone) and guinea pig anti-TRPV1 polyclonal (1:500, Alomone).

Techniques: Functional Assay, Cell Culture, Imaging, Expressing, MANN-WHITNEY

CB1r and P2X3r are Co-expressed in Lamina II of the Dorsal Horn of Mice and Naked Mole-Rat Spinal Cords Example confocal images of CB1r and P2X3r expression in the dorsal horn of the spinal cord in mice (A) and naked mole-rats (B). Scale bar 25 µm.

Journal: Neurobiology of Pain

Article Title: The naked mole-rat has a functional purinergic pain pathway despite having a non-functional peptidergic pain pathway

doi: 10.1016/j.ynpai.2020.100047

Figure Lengend Snippet: CB1r and P2X3r are Co-expressed in Lamina II of the Dorsal Horn of Mice and Naked Mole-Rat Spinal Cords Example confocal images of CB1r and P2X3r expression in the dorsal horn of the spinal cord in mice (A) and naked mole-rats (B). Scale bar 25 µm.

Article Snippet: Primary antibodies used were rabbit anti-P2X3r polyclonal (1:1000, Alomone) and guinea pig anti-TRPV1 polyclonal (1:500, Alomone).

Techniques: Expressing

SNL-induced changes in P2X3R expression in ipsilateral L4 and L5 DRG were altered by EA stimulation. (a) Examples of P2X3R expressions in different rat groups. (b) EA-induced changes in P2X3R protein expression in L4 DRGs. Following SNL+ s-EA stimulation, P2X3R expression was 1.4 times (= 1.1/0.77) higher than that in control rats. The expression was retuned close to control level (0.7/0.77 = 0.9) after EA stimulation. (c) EA-induced changes P2X3R protein expressions in L5 DRGs. The expression of P2X3R was much reduced after SNL+s-EA treatment (0.2/0.97 = 0.21) or after SNL+EA treatment (0.16/0.97 = 0.17). (n = 6 for each rat group, * P < 0.01). DRG: dorsal root ganglion; EA: electroacupuncture; GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

Journal: Molecular Pain

Article Title: Electroacupuncture downregulates P2X3 receptor expression in dorsal root ganglia of the spinal nerve-ligated rat

doi: 10.1177/1744806919847810

Figure Lengend Snippet: SNL-induced changes in P2X3R expression in ipsilateral L4 and L5 DRG were altered by EA stimulation. (a) Examples of P2X3R expressions in different rat groups. (b) EA-induced changes in P2X3R protein expression in L4 DRGs. Following SNL+ s-EA stimulation, P2X3R expression was 1.4 times (= 1.1/0.77) higher than that in control rats. The expression was retuned close to control level (0.7/0.77 = 0.9) after EA stimulation. (c) EA-induced changes P2X3R protein expressions in L5 DRGs. The expression of P2X3R was much reduced after SNL+s-EA treatment (0.2/0.97 = 0.21) or after SNL+EA treatment (0.16/0.97 = 0.17). (n = 6 for each rat group, * P < 0.01). DRG: dorsal root ganglion; EA: electroacupuncture; GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The samples were then electro-transferred to a polyvinylidene difluoride membrane, which was subsequently incubated in a Tris-buffered saline blocking buffer containing 5% w/v fat-free dry milk at room temperature (RT) for 1 h. The membrane was then immunoblotted with rabbit anti-P2X3R primary antibody (1:1000, Alomone Labs) at 4°C, overnight followed by horse radish peroxidase-conjugated goat anti-rabbit IgG (1:10000, Abcam) at RT for 1 h. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), probed with rabbit anti-GAPDH antibody (1:1000, Cell Signaling Technilogy), was used as loading control.

Techniques: Expressing

The effect of immunohistochemical study of P2X3R-ir and ATF3-ir expression in L4 DRG neurons. (a) P2X3R-ir (red) + ATF3-ir (green) double labeled DRG neurons in Control (Con), SNL+ s-EA (s-EA) and SNL+EA (EA) rats. Bar =100 µm. (b) Percentages of P2X3R+ expressing neurons in Control and SNL+ s-EA rats were similar (28.24 ± 1.69% in Con vs. 26.83 ± 0.9% in s-EA rats), The percentage of P2X3R+ neurons in SNL+ EA rats was significantly reduced (15.37 ± 1.75%). (c) Integrated optical density measured in SNL+s-EA rats ((9.21 ± 1.08) × 10 5 ) was higher than that in control rats ((6.64 ± 0.75) ×10 5 ) and much reduced ((2.81 ± 0.39) × 10 5 ) in SNL+EA rats. (d) The percentage of ATF+ neurons in SNL+ s-EA and in SNL+EA rats were not significantly different (6.7 ± 1.7 vs. 6.1 ± 0.6%, P > 0.05). Both were higher than control rats (0%). (e) Almost no P2X3+/ATF3+ neurons were expressed in the three rat groups. Thus, all of the P2X3R+ were expressed in un-injured neurons (Con: n = 7, SNL+ s-EA: n = 3, SNL+EA: n = 3). * P < 0.01. EA: electroacupuncture; ATF3: activating transcription factor 3; IOD: integrated optical density.

Journal: Molecular Pain

Article Title: Electroacupuncture downregulates P2X3 receptor expression in dorsal root ganglia of the spinal nerve-ligated rat

doi: 10.1177/1744806919847810

Figure Lengend Snippet: The effect of immunohistochemical study of P2X3R-ir and ATF3-ir expression in L4 DRG neurons. (a) P2X3R-ir (red) + ATF3-ir (green) double labeled DRG neurons in Control (Con), SNL+ s-EA (s-EA) and SNL+EA (EA) rats. Bar =100 µm. (b) Percentages of P2X3R+ expressing neurons in Control and SNL+ s-EA rats were similar (28.24 ± 1.69% in Con vs. 26.83 ± 0.9% in s-EA rats), The percentage of P2X3R+ neurons in SNL+ EA rats was significantly reduced (15.37 ± 1.75%). (c) Integrated optical density measured in SNL+s-EA rats ((9.21 ± 1.08) × 10 5 ) was higher than that in control rats ((6.64 ± 0.75) ×10 5 ) and much reduced ((2.81 ± 0.39) × 10 5 ) in SNL+EA rats. (d) The percentage of ATF+ neurons in SNL+ s-EA and in SNL+EA rats were not significantly different (6.7 ± 1.7 vs. 6.1 ± 0.6%, P > 0.05). Both were higher than control rats (0%). (e) Almost no P2X3+/ATF3+ neurons were expressed in the three rat groups. Thus, all of the P2X3R+ were expressed in un-injured neurons (Con: n = 7, SNL+ s-EA: n = 3, SNL+EA: n = 3). * P < 0.01. EA: electroacupuncture; ATF3: activating transcription factor 3; IOD: integrated optical density.

Techniques: Immunohistochemical staining, Expressing, Labeling

EA stimulation did not affect P2X3R or ATF3 expression in injured neurons ipsilateral to L5 DRGs. (a) Examples of P2X3R-ir + ATF3-ir double labeled neurons in L5 DRGs of three rat groups, Bar = 100 µm. (b) Compared to control rats, L5 nerve ligation induced a substantial reduction in P2X3R+ expression in DRGs of SNL+ s-EA or SNL+EA rats (from 29.0 ± 2.1% to 13.7 ± 4.9% and to 11.0 ± 3.3% respectively). (c) Unlike the minimal expression of ATF3+ cells (0.4 ± 0.2%) in L5 DRG of control rats, a majority of L5 DRG neurons in SNL+ s-EA rat group (85.7 ± 1.6%) and in SNL+EA group (81.7 ± 3.1%) were ATF3+. (d) Most P2X3Rs in injured L5 DRGs were expressed in ATF3+ neurons (P2X3R/ATF3+ =10.6% in SNL+ s-EA rats and =8.8% in SNL+EA rats). (e) The cell count of P2X3R+/ATF3− cells was reduced from 30.0 ± 2.0% in control L5 DRGs to 2.7 ± 0.8% in SNL+ s-EA and 1.8 ± 0.5% in SNL+EA rats. EA stimulation did not reverse the reduction in P2X3R expression induced by SNL (Con: n = 7, SNL+ s-EA: n = 3, SNL+EA: n = 3). * P < 0.01. EA: electroacupuncture; ATF3: activating transcription factor 3.

Journal: Molecular Pain

Article Title: Electroacupuncture downregulates P2X3 receptor expression in dorsal root ganglia of the spinal nerve-ligated rat

doi: 10.1177/1744806919847810

Figure Lengend Snippet: EA stimulation did not affect P2X3R or ATF3 expression in injured neurons ipsilateral to L5 DRGs. (a) Examples of P2X3R-ir + ATF3-ir double labeled neurons in L5 DRGs of three rat groups, Bar = 100 µm. (b) Compared to control rats, L5 nerve ligation induced a substantial reduction in P2X3R+ expression in DRGs of SNL+ s-EA or SNL+EA rats (from 29.0 ± 2.1% to 13.7 ± 4.9% and to 11.0 ± 3.3% respectively). (c) Unlike the minimal expression of ATF3+ cells (0.4 ± 0.2%) in L5 DRG of control rats, a majority of L5 DRG neurons in SNL+ s-EA rat group (85.7 ± 1.6%) and in SNL+EA group (81.7 ± 3.1%) were ATF3+. (d) Most P2X3Rs in injured L5 DRGs were expressed in ATF3+ neurons (P2X3R/ATF3+ =10.6% in SNL+ s-EA rats and =8.8% in SNL+EA rats). (e) The cell count of P2X3R+/ATF3− cells was reduced from 30.0 ± 2.0% in control L5 DRGs to 2.7 ± 0.8% in SNL+ s-EA and 1.8 ± 0.5% in SNL+EA rats. EA stimulation did not reverse the reduction in P2X3R expression induced by SNL (Con: n = 7, SNL+ s-EA: n = 3, SNL+EA: n = 3). * P < 0.01. EA: electroacupuncture; ATF3: activating transcription factor 3.

Techniques: Expressing, Labeling, Ligation, Cell Counting

CPT increases the membrane expression of P2X3Rs in cultured DRGs. The P2X3R levels in total protein (Total), membrane protein (Mem), and cytoplasmic protein samples (Cytosol) before (Con) and after CPT (1 µM, 10 min) treatment were determined. Total, membrane, and cytosol protein samples were loaded onto gels. The expression of the ubiquitous membrane protein Na+/K+ ATPase was probed to indicate the membrane expressed protein levels in our samples. The expressions of β-tubulin used as sample loading controls were also indicated. CPT significantly increased P2X3Rs expressed in the cell membrane (total protein: CPT/Con = 1.19; membrane protein: CPT/Con = 1.93 and cytoplasmic protein: CPT/Con = 0.75). N = 3. * P < 0.05.

Journal: Molecular Pain

Article Title: F-actin links Epac-PKC signaling to purinergic P2X3 receptor sensitization in dorsal root ganglia following inflammation

doi: 10.1177/1744806916660557

Figure Lengend Snippet: CPT increases the membrane expression of P2X3Rs in cultured DRGs. The P2X3R levels in total protein (Total), membrane protein (Mem), and cytoplasmic protein samples (Cytosol) before (Con) and after CPT (1 µM, 10 min) treatment were determined. Total, membrane, and cytosol protein samples were loaded onto gels. The expression of the ubiquitous membrane protein Na+/K+ ATPase was probed to indicate the membrane expressed protein levels in our samples. The expressions of β-tubulin used as sample loading controls were also indicated. CPT significantly increased P2X3Rs expressed in the cell membrane (total protein: CPT/Con = 1.19; membrane protein: CPT/Con = 1.93 and cytoplasmic protein: CPT/Con = 0.75). N = 3. * P < 0.05.

Article Snippet: Immediately after treatment, DRG cells were fixed with 4% paraformaldehyde at room temperature for 20 min, washed with phosphate-buffered saline (PBS), and blocked with PBS containing 5% normal goat serum for 30 min. To determine the membrane expression of P2X3Rs in DRG cells, the primary antibody rabbit anti-P2X3R extracellular epitope, (ext-P2X3R), (1:100, Alomone Lab, Jerusalem), was used.

Techniques: Expressing, Cell Culture

Epac control of the membrane expression of P2X3Rs in DRG neurons is F-actin mediated. (a) The membrane expression of P2X3Rs was examined in cultured DRG neurons using anti-ext-P2X3R antibody. The enlarged views of neurons (indicated by red arrows) are shown on the right side of each panel. Bars = 25 µm. (b) In the presence of CPT (1 µM), the percentage of cells expressing ext-P2X3R increased by 1.80 fold and the intensity of ext-P2X3R labels increased by 5.57 fold. The CPT-induced increase in membrane expressed P2X3Rs was significantly reduced by pre-incubating cells with the F-actin disrupter, LaA (1 µM, 60 min). A total of 1192 cells obtained from three experiments were used for the analyses. * P < 0.05, # P < 0.01. NS: not significant.

Journal: Molecular Pain

Article Title: F-actin links Epac-PKC signaling to purinergic P2X3 receptor sensitization in dorsal root ganglia following inflammation

doi: 10.1177/1744806916660557

Figure Lengend Snippet: Epac control of the membrane expression of P2X3Rs in DRG neurons is F-actin mediated. (a) The membrane expression of P2X3Rs was examined in cultured DRG neurons using anti-ext-P2X3R antibody. The enlarged views of neurons (indicated by red arrows) are shown on the right side of each panel. Bars = 25 µm. (b) In the presence of CPT (1 µM), the percentage of cells expressing ext-P2X3R increased by 1.80 fold and the intensity of ext-P2X3R labels increased by 5.57 fold. The CPT-induced increase in membrane expressed P2X3Rs was significantly reduced by pre-incubating cells with the F-actin disrupter, LaA (1 µM, 60 min). A total of 1192 cells obtained from three experiments were used for the analyses. * P < 0.05, # P < 0.01. NS: not significant.

Techniques: Expressing, Cell Culture

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