Journal: Biochimica et biophysica acta

Article Title: Prophylactic systemic P2X7 receptor blockade prevents experimental colitis.

doi: 10.1016/j.bbadis.2013.10.012

Figure Lengend Snippet: Fig. 8. Effect of P2X7-R blockade with A740003 on the colonoscopic parameters and histological parameters in the colitis model. Colonoscopic imaging was obtained after colitis induction at day 7. In control experiments, colonoscopy was performed following intra-rectal saline enemas, and intraperitoneal administration of vehicle in TNBS-induced animals. Animals treated with intraperitoneal (IP) A740003, showed significantly less inflammation as assessed by colonoscopy, in contrast to vehicle-treated animals (p = 0.005). Values are mean ± S.E.M. of 5–6 animals per group. Significant values are presented (A). Paraffin sections were stained with HE. Intraperitoneal (IP)-A740003 was administered at 16 mg/kg at day –1. In control experi- ments, histological evaluation was performed following intraperitoneal administration of vehicle, and intra-rectal saline enemas. Colonic samples were scored according to histological pa- rameters described in the Materials and methods section. Animals treated with IP-A740003 showed significantly less inflammation measured by histologic parameters compared to vehicle-treated animals (p = 0.034). Values are mean ± S.E.M. of 5–6 animals per group. Significant values are presented (B). Length bars represent 20 μm.

Article Snippet: The P2X7-R blockers utilized in this work were the brilliant blue G (BBG) (Sigma-Aldrich, Saint Louis, MO, USA) and A740003 [N-(1-{[(cyanoimino)(5-quinolinylamino) methyl] amino}-2,2-dimethylpropyl)-2-(3,4-dimethoxyphenyl) acetamide] (Tocris Bioscience, Bristol, UK) [35].

Techniques: Imaging, Control, Saline, Staining

Journal: The Journal of Headache and Pain

Article Title: Src family kinases activity is required for transmitting purinergic P2X7 receptor signaling in cortical spreading depression and neuroinflammation

doi: 10.1186/s10194-021-01359-8

Figure Lengend Snippet: Both deactivation of P2X7 receptor and SFKs reduced cortical susceptibility to CSD in mouse brain slices. a The captured images of a mouse brain slice before and after CSD induction by 260 mM KCl in cerebral cortex. The arrow indicated the direction of CSD propagation. An AOI was selected and kept the same for data analysis. b The biphasic CSD curve generated from the images recorded for 15 minutes by plotting averaged grey level within the AOI against time. CSD latency (sec) is the time interval between KCl application and CSD elicitation at the AOI. CSD propagation rate (mm/min) is the velocity by which CSD propagates along cerebral cortex. c Representative traces of the 1st peak of CSD affected by 0.03% DMSO, 3 μM A740003 and 0.5 μM saracatinib. Only the trace recorded during the first 100 seconds was displayed here in order to clearly show the starting points of KCl application and CSD elicitation. d e Effects of 0.03% DMSO, 3 μM A740003 and 0.5 μM saracatinib on CSD latency and propagation rate. Abbreviations: saracatinib (SRCT); seconds (sec); mm/minute (mm/min). Two-tailed unpaired t-test was used for comparison in CSD latency and propagation rate between DMSO and A740003 group, DMSO and saracatinib group. Significant differences were indicated by * p < 0.05, ** p < 0.01

Article Snippet: Three series of studies were designed using this ex vivo CSD model. Series 1: In order to study whether modulation of SFKs or P2X7 receptor activity could similarly regulate cortical susceptibility to CSD, the effects of a SFKs inhibitor, saracatinib (S1006, Selleckchem, Houston, USA), that is currently investigated in clinical trials for treating several neurological disorders [ , ] or a P2X7 receptor antagonist, A740003 (3701, Tocris, Bristol, UK) on CSD latency and propagation rate were examined respectively in C57BL/6J mice brain slices.

Techniques: Slice Preparation, Generated, Two Tailed Test, Comparison

Journal: The Journal of Headache and Pain

Article Title: Src family kinases activity is required for transmitting purinergic P2X7 receptor signaling in cortical spreading depression and neuroinflammation

doi: 10.1186/s10194-021-01359-8

Figure Lengend Snippet: Disrupting P2X7 receptor-SFKs interaction reduced cortical susceptibility to CSD and CSD-associated neuroinflammation in mouse brain slices. a Representative traces of 1 st peak of CSD affected by Kreb’s, 3 μM TAT-P2X7 and TAT-P2X7SC. Only the trace recorded during the first 100 seconds was displayed here in order to clearly show CSD latency in respective group. b c Effects of Kreb’s, 3 μM TAT-P2X7 and TAT-P2X7SC on CSD latency and propagation rate. d e Effects of 3 μM TAT-P2X7 and TAT-P2X7SC on IL-1β and TNFα mRNA fold change after CSD in mouse brain slices. Two-tailed unpaired t-test was used for comparison in CSD latency and propagation rate between Kreb’s and TAT-P2X7 group, TAT-P2X7 and TAT-P2X7SC group; in IL-1β and TNFα mRNA fold change between TAT-P2X7 and TAT-P2X7SC group. Significant differences were indicated by * p < 0.05, ** p < 0.01

Article Snippet: Three series of studies were designed using this ex vivo CSD model. Series 1: In order to study whether modulation of SFKs or P2X7 receptor activity could similarly regulate cortical susceptibility to CSD, the effects of a SFKs inhibitor, saracatinib (S1006, Selleckchem, Houston, USA), that is currently investigated in clinical trials for treating several neurological disorders [ , ] or a P2X7 receptor antagonist, A740003 (3701, Tocris, Bristol, UK) on CSD latency and propagation rate were examined respectively in C57BL/6J mice brain slices.

Techniques: Two Tailed Test, Comparison

Journal: The Journal of Headache and Pain

Article Title: Src family kinases activity is required for transmitting purinergic P2X7 receptor signaling in cortical spreading depression and neuroinflammation

doi: 10.1186/s10194-021-01359-8

Figure Lengend Snippet: NMDA restored the disrupted P2X7 receptor-SFKs interaction-reduced cortical susceptibility to CSD but not CSD-associated neuroinflammation in mouse brain slices. a Effects of 3 μM TAT-P2X7 and TAT-P2X7SC on glutamate release from mouse brain slices after CSD. b Representative traces of 1 st peak of CSD affected by 3 μM TAT-P2X7 and 3 μM TAT-P2X7 + 10 μM NMDA. c d Effects of 3 μM TAT-P2X7 + 10 μM NMDA on CSD latency and propagation rate. e Effects of 3 μM TAT-P2X7 + 10 μM NMDA on CSD-associated IL-1β mRNA fold change in mouse brain slices. Two-tailed unpaired t-test was used for comparison in glutamate release between TAT-P2X7 and TAT-P2X7SC group; in CSD latency, propagation rate and IL-1β mRNA fold change between TAT-P2X7 and TAT-P2X7 + NMDA group. Significant differences were indicated by * p < 0.05

Article Snippet: Three series of studies were designed using this ex vivo CSD model. Series 1: In order to study whether modulation of SFKs or P2X7 receptor activity could similarly regulate cortical susceptibility to CSD, the effects of a SFKs inhibitor, saracatinib (S1006, Selleckchem, Houston, USA), that is currently investigated in clinical trials for treating several neurological disorders [ , ] or a P2X7 receptor antagonist, A740003 (3701, Tocris, Bristol, UK) on CSD latency and propagation rate were examined respectively in C57BL/6J mice brain slices.

Techniques: Two Tailed Test, Comparison

Journal: The Journal of Headache and Pain

Article Title: Src family kinases activity is required for transmitting purinergic P2X7 receptor signaling in cortical spreading depression and neuroinflammation

doi: 10.1186/s10194-021-01359-8

Figure Lengend Snippet: Schematic representation of the role that P2X7 receptor/SFKs signaling may exert in CSD-associated migraine pathophysiology. P2X7 receptor/SFKs signaling is activated during CSD to facilitate neuroinflammation, NMDA receptor activation and glutamate release. Glutamate may in return reinforce the activation of NMDA receptor (dotted line with arrow), both of which facilitate cortical susceptibility to CSD, forming a positive loop

Article Snippet: Three series of studies were designed using this ex vivo CSD model. Series 1: In order to study whether modulation of SFKs or P2X7 receptor activity could similarly regulate cortical susceptibility to CSD, the effects of a SFKs inhibitor, saracatinib (S1006, Selleckchem, Houston, USA), that is currently investigated in clinical trials for treating several neurological disorders [ , ] or a P2X7 receptor antagonist, A740003 (3701, Tocris, Bristol, UK) on CSD latency and propagation rate were examined respectively in C57BL/6J mice brain slices.

Techniques: Activation Assay

Journal: Molecular Pain

Article Title: Activation of the P2X7 receptor in the dental pulp tissue contributes to the pain in rats with acute pulpitis

doi: 10.1177/17448069221106844

Figure Lengend Snippet: Immunohistochemical staining of the dental pulp of rat. Normal group: A1 crown pulp; A2 upper 1/3 of the root; A3 middle 1/3 of the root; and A4 root tip 1/3; NS group: B1 crown pulp; B2 upper 1/3 of the root; B3 middle 1/3 of the root; and B4 root tip 1/3; LPS group: C1 crown pulp; C2 upper 1/3 of the root; C3 middle 1/3 of the root; and C4 root tip 1/3; Transverse images of the mesial root were selected for the pulp parts, the arrows indicate the P2X7 receptor expressed in the odontoblast layer in a yellowish-brown granular form. Scale bar = 50 μm.

Article Snippet: Escherichia coli lipopolysaccharide (LPS) was purchased from Sigma, USA (L2880, 1 mg/mL, diluted in NS); Rabbit polyclonal antibody against rat P2X7 was bought from Abcam (USA); Rabbit Hypersensitivity two-step Detection Kit, bovine serum protein, and 3,3’-diaminobenzidine (DAB) color Development Kit were acquired from Beijing Zhongshan Jinqiao; P2X7 receptor antagonist A-740003 was procured from TargetMol (USA).

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Pain

Article Title: Activation of the P2X7 receptor in the dental pulp tissue contributes to the pain in rats with acute pulpitis

doi: 10.1177/17448069221106844

Figure Lengend Snippet: Western Blotting bands of the P2X7 receptor protein in the dental pulp tissue of rats in each group.

Article Snippet: Escherichia coli lipopolysaccharide (LPS) was purchased from Sigma, USA (L2880, 1 mg/mL, diluted in NS); Rabbit polyclonal antibody against rat P2X7 was bought from Abcam (USA); Rabbit Hypersensitivity two-step Detection Kit, bovine serum protein, and 3,3’-diaminobenzidine (DAB) color Development Kit were acquired from Beijing Zhongshan Jinqiao; P2X7 receptor antagonist A-740003 was procured from TargetMol (USA).

Techniques: Western Blot

Journal: Molecular Pain

Article Title: Activation of the P2X7 receptor in the dental pulp tissue contributes to the pain in rats with acute pulpitis

doi: 10.1177/17448069221106844

Figure Lengend Snippet: Comparison of the P2X7 receptor protein expression in the dental pulp tissue of rats in each group. *** p < 0.001, compared to the normal group; and ▲▲▲ p < 0.001, compared to the NS group.

Article Snippet: Escherichia coli lipopolysaccharide (LPS) was purchased from Sigma, USA (L2880, 1 mg/mL, diluted in NS); Rabbit polyclonal antibody against rat P2X7 was bought from Abcam (USA); Rabbit Hypersensitivity two-step Detection Kit, bovine serum protein, and 3,3’-diaminobenzidine (DAB) color Development Kit were acquired from Beijing Zhongshan Jinqiao; P2X7 receptor antagonist A-740003 was procured from TargetMol (USA).

Techniques: Expressing