Journal: Molecular Pain

Article Title: Comparison of P2X and TRPV1 receptors in ganglia or primary culture of trigeminal neurons and their modulation by NGF or serotonin

doi: 10.1186/1744-8069-2-11

Figure Lengend Snippet: Calcium imaging indicates functional expression of P2X and TRPV1 receptors . A, examples of Ca 2+ transients in mouse TG neurons activated by 2 s application of 10 μM α, β-meATP or 1 μM capsaicin. a, example of cell responding to α, β-meATP only ('purinergic phenotype'); b, responding to capsaicin only ('vanilloid phenotype'); c, responding to both agonists ('mixed phenotype'). All cells respond to pulse application of KCl. B, number of neurons sensitive to α, β-meATP (regardless of their response to capsaicin), to capsaicin (regardless of their response to α, β-meATP) or to both agonists, for mouse (n = 346; left) and rat (n = 41; right) TG neurons. Neurons (kept in culture for 24 h) were identified by their responsiveness to 15 mM KCl (1 s), while satellite cells and fibroblasts did not respond to this agent. Note that the percent of cells responding to capsaicin was higher in the rat (55%) than in the mouse (33%).

Article Snippet: α, β-meATP, NGF and capsaicin (all from Sigma) were diluted with physiological solution to final concentration and applied by a rapid superfusion system (Rapid Solution Changer RSC-200, BioLogic Science Instruments, Grenoble, France).

Techniques: Imaging, Functional Assay, Expressing

Journal: Molecular Pain

Article Title: Comparison of P2X and TRPV1 receptors in ganglia or primary culture of trigeminal neurons and their modulation by NGF or serotonin

doi: 10.1186/1744-8069-2-11

Figure Lengend Snippet: Characteristics of pain receptors of cultured TG neurons . A, examples of currents recorded from rat and mouse TG neurons in culture (24 h) during application of α, β-meATP (10 μM, 2 s; upper row) and capsaicin (10 μM, 2 s for rat; 1 μM, 2 s for mouse; bottom row). B, log dose-response curves for rat (filled symbols; n = 9) and mouse (grey symbols; n = 12) neurons cultured for 24 h. Both potency and efficacy of α, β-meATP were similar for rat and mouse. C, distribution of values of the residual current (I residual ) present at the end of an application of α, β-meATP (10 μM, 2 s) in rat and mouse neurons cultured for 24 h, expressed as a fraction of the peak current (I peak ; n = 24 and 58 for rat and mouse, respectively). Inset shows an example of the mixed type of current recorded from a subset of rat TG neurons. Dashed line indicates the 5% arbitrary threshold above which currents were considered to be mixed.

Article Snippet: α, β-meATP, NGF and capsaicin (all from Sigma) were diluted with physiological solution to final concentration and applied by a rapid superfusion system (Rapid Solution Changer RSC-200, BioLogic Science Instruments, Grenoble, France).

Techniques: Cell Culture

Journal: Molecular Pain

Article Title: Comparison of P2X and TRPV1 receptors in ganglia or primary culture of trigeminal neurons and their modulation by NGF or serotonin

doi: 10.1186/1744-8069-2-11

Figure Lengend Snippet: Functional characterization of rat and mouse TG neurons . A, fraction of cells responding to α, β-meATP (independently from their response to capsaicin), to capsaicin (independently from their response to α, β-meATP) or to both agonists for rat (left panel; n = 33) and mouse (right panel; n = 50). B, persistence of the responsiveness of rat TG neurons in culture to α, β-meATP and capsaicin. a, Proportion of cells responding to α, β-meATP (black bars; n = 7, 7, 10 cell cultures for the 1 st , 2 nd and 3 rd day, respectively) and capsaicin (grey bars; n = 7, 8 and 8 cell cultures for the 1 st , 2 nd and 3 rd day). b, peak amplitude of currents elicited by α, β-meATP (black bars; n = 22, 11 and 31 cells for the 1 st , 2 nd and 3 rd day, respectively) and capsaicin (grey bars; n = 16, 13 and 10 cells for the 1 st , 2 nd and 3 rd day). Only cells responding to the agonist were included in the current analysis.

Article Snippet: α, β-meATP, NGF and capsaicin (all from Sigma) were diluted with physiological solution to final concentration and applied by a rapid superfusion system (Rapid Solution Changer RSC-200, BioLogic Science Instruments, Grenoble, France).

Techniques: Functional Assay

Journal: Molecular Pain

Article Title: Comparison of P2X and TRPV1 receptors in ganglia or primary culture of trigeminal neurons and their modulation by NGF or serotonin

doi: 10.1186/1744-8069-2-11

Figure Lengend Snippet: Modulation of receptor function by chronically applied NGF or 5-HT . A, Ca 2+ imaging of single neurons shows percent increase in mouse TG cells responsive to α, β-meATP (a) in control or after application of 50 ng/ml NGF (24 h; n = 6 culture dishes). In 6 sister cultures there was no significant change as far as responses to capsaicin (b) were concerned. *: P < 0.05. B, a, patch clamp current records show increased amplitude of mouse responses to α, β-meATP after 24 h NGF treatment, while responses to capsaicin remained equiamplitude. B, b, histograms summarizing the significant (*: P < 0.05) rise in α, β-meATP evoked current amplitude (n = 26) without significant change in capsaicin responses (n = 14). Data are expressed as % of control amplitude in sister cultures. C, 5-HT (10 μM; 24 h) upregulates the amplitude of rat capsaicin current without affecting responses to α, β-meATP (a). C,b shows significant rise in the peak current induced by capsaicin (n = 31) with no change in the α, β-meATP-evoked current (n = 31). Data are expressed as % of control amplitude in sister cultures. *: P < 0.05.

Article Snippet: α, β-meATP, NGF and capsaicin (all from Sigma) were diluted with physiological solution to final concentration and applied by a rapid superfusion system (Rapid Solution Changer RSC-200, BioLogic Science Instruments, Grenoble, France).

Techniques: Imaging, Patch Clamp

Journal: Respiratory Research

Article Title: Acute lung inflammation and ventilator-induced lung injury caused by ATP via the P2Y receptors: an experimental study

doi: 10.1186/1465-9921-9-79

Figure Lengend Snippet: Lung inflammation caused by P2Y and P2X selective agonist . Hematoxilin-eosin staining of the lung 24 h after the instillation of UTP or α,β-MeATP. Infiltration of inflammatory cells, increased thickness of the alveolar walls and alveolar hemorrhages were observed in the 200 mM UTP-treated lungs. However, there was no evident derangement in the 200 mM α,β-MeATP-treated lungs. Bars = 100 μm in the upper panels, and = 50 μm in the lower panels. B. Immunohistochemistry of the P2Y 2 and P2Y 4 receptors. P2Y 2 and P2Y 4 receptors were detected in bronchiolar epithelial cells, alveolar walls and alveolar macrophages in the lung of untreated mice. Bars = 100 μm in the upper panels, and = 50 μm in the lower panels.

Article Snippet: Adenosine 5'-triphosphate (ATP), selective P2Xs, P2Y 2 and P2Y 4 antagonist pyridoxal-5'-phosphate-6-azophenyl-2', 4 '-disulfonic acid (PPADS), selective P2Y agonist uridine 5'-triphosphate (UTP) and selective P2X agonist α,β-methylene ATP (α,β-MeATP) were obtained from Sigma-Aldrich (St. Louis, MO).

Techniques: Staining, Immunohistochemistry