Journal: Molecular Pain

Article Title: Acute pulpitis promotes purinergic signaling to induce pain in rats via P38MAPK/NF-κB signaling pathway

doi: 10.1177/17448069241234451

Figure Lengend Snippet: LPS induced upregulation of P2X3 expression in the ipsilateral TG. (a) Representative pictures of immunohistochemical labelling for P2X3 in the TG at different time points in pulpitis rats. Some neurons with P2X3 immunopositive nuclei are marked by arrowheads. Red arrows mark neurons with small diameters, blue arrows mark neurons with medium diameters (Bar = 50 μm). (b) The percentage of immunohistochemically positive area of p2x3 expressed in TG was quantitatively analysed. (c) Representative immunoblots of samples from rat TGs subjected to pulpitis model with quantitative densitometric analysis of P2X3 protein with β-actin as an internal standard. The immunoblots were obtained from the microgel running under the same experimental conditions. (d) Graphical representation of P2X3 mRNA expression in TGs. All the data are normalised to the left TG of the SHAM group. ** p < .01 versus SHAM group; ## p < .01 versus LPS-1 group; && p < .01 versus LPS-3 group; n = 6 for each group.

Article Snippet: The rats in the LPS-1A and LPS-3A groups were injected intraperitoneally with the selective P2X3 inhibitor A-317491 (Sigma, St Louis, Missouri, USA; 0.5 mg/kg, dissolved in 0.9% NaCl) immediately after LPS application, and scarified after 1 day and 3 days respectively.

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Molecular Pain

Article Title: Acute pulpitis promotes purinergic signaling to induce pain in rats via P38MAPK/NF-κB signaling pathway

doi: 10.1177/17448069241234451

Figure Lengend Snippet: A Immunofluorescent staining of c-fos and P2X3 expression in the trigeminal ganglion. Representative photomicrographs of c-fos and P2X3 expression in 5 groups of rats. In the control group, c-fos was only slightly expressed in the TG, and almost no P2X3-positive neurons were found. The expression of c-fos in the LPS-1 group and LPS-3 group was significantly increased, and c-fos and P2X3 were coexpressed in neurons, the c-fos and P2X3 co-positive neurons were marked by white arrowheads. Scale bar: 100 µm. B Changes in the mean percentage of P2X3-IR and TNFR2-IR TG neurons on day1 and day3 following pulpitis. * p < .05 versus SHAM. ( n = 6 in each).

Article Snippet: The rats in the LPS-1A and LPS-3A groups were injected intraperitoneally with the selective P2X3 inhibitor A-317491 (Sigma, St Louis, Missouri, USA; 0.5 mg/kg, dissolved in 0.9% NaCl) immediately after LPS application, and scarified after 1 day and 3 days respectively.

Techniques: Staining, Expressing

Journal: Frontiers in Physiology

Article Title: Piezo1-pannexin-1-P2X 3 axis in odontoblasts and neurons mediates sensory transduction in dentinal sensitivity

doi: 10.3389/fphys.2022.891759

Figure Lengend Snippet: Time-dependence of pharmacological effects of P2X 3 receptor (A-317491) and PANX (mefloquine and 10 PANX) antagonist on dentinal sensitivity (A–C) Time-dependent effects of A-317491 (A; n = 6, 0.3 nmol/g (weight) i.v.), mefloquine (B; n = 7, 0.03 mg/g (weight) p.o.), and 10 PANX (C; n = 8, 2.58 nmol/g (weight) i.v.) (upper white boxes) on nociceptive scores following cold water stimulation. Prior to the systemic administration of A-317491, mefloquine, or 10 PANX to the dentin-exposed rats, nociceptive scores were determined as the control values (white circles). Several minutes after these measurements, A-317491 (A) , mefloquine (B) , or 10 PANX (C) was administered to the rats. The cold water stimuli-induced nociceptive scores were measured repeatedly at 10, 60, 120, 180, 240, 300, and 360 min later each (black circles in A to C; see also Materials and Methods). Each point denotes the mean ± SE of the number of experiments. Significant differences in the scores compared to control values are denoted by asterisks: * p < 0.05. (D) Summary bar graphs showing the mean nociceptive scores in the dentin-exposed group of rats without any treatment (control; withe columns) and their minimum values (during the entire period of administration) in dentin-exposed rats treated with A-317491 ( n = 6), mefloquine ( n = 7) and 10 PANX ( n = 8) (black columns). Each bar represents mean ± SE of the number of experiments. Significant differences between columns (shown by solid lines) are denoted by asterisks: * p < 0.05.

Article Snippet: The stock solutions of the reagents used included A-317491, a P2X 3 receptor antagonist (Sigma-Aldrich); 10 PANX, a selective PANX-1 blocker; and GsMTx-4, a selective Piezo1 channel blocker (R&D Systems, Inc, Minneapolis, United States) were prepared in sterilized PBS (Life Technologies, Ltd, Paisley, United Kingdom) and diluted with physiological saline (Otsuka Pharmaceutical Co., Ltd., Tokyo, Japan) to the appropriate concentration for administration.

Techniques:

Journal: Frontiers in Physiology

Article Title: Piezo1-pannexin-1-P2X 3 axis in odontoblasts and neurons mediates sensory transduction in dentinal sensitivity

doi: 10.3389/fphys.2022.891759

Figure Lengend Snippet: Pharmacological effects of Piezo1 channel (GsMTx-4), PANX-1 ( 10 PANX) and P2X 3 receptor (A-317491) antagonists on dentinal sensitivity in rat first molar (A–C) Bar graphs showing the mean nociceptive scores following cold water (0.1 ml, 4.0–7.0°C) stimulation to the dentin surface of the lower first molar in the dentin-exposed group rats without any treatment (control; withe columns) and their values treated with GsMTx-4 (A; n = 6), A-317491 (B; n = 6), and 10 PANX (C; n = 6) (black columns) 60 min (middle columns) and 120 min (lower columns) after administration of each agent. The cold water stimuli-induced nociceptive scores were measured repeatedly at 60 and 120 min later each (see also Materials and Methods). Each bar represents the mean ± SE of the number of experiments. Asterisks denote significant differences between columns (shown by solid lines): * p < 0.05.

Article Snippet: The stock solutions of the reagents used included A-317491, a P2X 3 receptor antagonist (Sigma-Aldrich); 10 PANX, a selective PANX-1 blocker; and GsMTx-4, a selective Piezo1 channel blocker (R&D Systems, Inc, Minneapolis, United States) were prepared in sterilized PBS (Life Technologies, Ltd, Paisley, United Kingdom) and diluted with physiological saline (Otsuka Pharmaceutical Co., Ltd., Tokyo, Japan) to the appropriate concentration for administration.

Techniques:

Journal: Frontiers in Physiology

Article Title: Piezo1-pannexin-1-P2X 3 axis in odontoblasts and neurons mediates sensory transduction in dentinal sensitivity

doi: 10.3389/fphys.2022.891759

Figure Lengend Snippet: Electrophysiological recordings from trigeminal ganglion (TG) neurons (A) Representative trace (upper) and its spike histogram showing spike number in every 5 s (lower) of event-related neuronal activities from the whole TG neurons following cold water (middle black box) or warm water (middle open boxes) stimulation to exposed dentin surface of lower first molars in rat injected by physiological saline intraperitoneally. The stimulated dentin was adapted to warm (37.0°C) distilled water, and the cold (4.0–7.0°C) stimulus was applied for ∼30 s (B) Averaged spike frequencies in response to cold water stimuli to exposed dentin surface of lower first molars in the rat following intraperitoneal injection of physiological saline (upper; n = 6), A-317491 (second upper; n = 5), GsMTx-4 (second lower; n = 5) or 10 PANX (lower; n = 5). Each bar represents the mean ± SD of the number of experiments. Asterisks denote significant differences between columns (shown by solid lines): * p < 0.05.

Article Snippet: The stock solutions of the reagents used included A-317491, a P2X 3 receptor antagonist (Sigma-Aldrich); 10 PANX, a selective PANX-1 blocker; and GsMTx-4, a selective Piezo1 channel blocker (R&D Systems, Inc, Minneapolis, United States) were prepared in sterilized PBS (Life Technologies, Ltd, Paisley, United Kingdom) and diluted with physiological saline (Otsuka Pharmaceutical Co., Ltd., Tokyo, Japan) to the appropriate concentration for administration.

Techniques: Injection