Journal: Frontiers in Molecular Neuroscience

Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

doi: 10.3389/fnmol.2017.00042

Figure Lengend Snippet: PAR2-activation induces a depression of synaptic transmission at Schaffer collateral-CA1 synapses in the hippocampus. (A) Application of PAR2-agonist (10 μM AC55541) causes LTD. (B) Removal of the PAR2-agonist (10 μM AC55541) following induction of LTD does not affect the stability of synaptic depression. (C) In presence of a PAR2-antagonist (50 μM FSLLRY-NH 2 ) the PAR2-agonist (10 μM AC55541) is not able to induce synaptic depression. (D) Application of PAR2-agonist (10 μM AC55541) at different concentrations results in similar levels of synaptic depression. (E) In a two pathways experimental setting, low frequency stimulation (LFS, 1 Hz, 900 pulses) and PAR2-activation (10 μM AC55541) induce similar levels of LTD. (F) LFS-induced LTD is not blocked by the PAR2-antagonist. (G) In a two pathways experimental setting, the NMDAR-antagonist (50 μM APV) blocks both LFS-induced LTD and PAR2-induced LTD. (H) While the group I mGluR-antagonist MCPG (200 μM) partially affects LFS-LTD it does not influence PAR2-LTD. Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section, n = 12 slices for each experiment, refer to text for statistics.

Article Snippet: The following compounds were used at the following concentrations: 10 μM AC55541 (PAR2-agonist, Tocris Bioscience, UK), 10 μM AC264613 (PAR2-agonist, Tocris Bioscience, UK), 50 μM FSLLRY-NH 2 (PAR2-antagonist, Sigma-Aldrich, Israel), 2 μM RN1747 (TRPV4-agonist, Tocris Bioscience, UK), 10 μM RN1734 (TRPV4-antagonist, Tocris Bioscience, UK), 10 μM RN9893 (TRPV4-antagonist, Tocris Bioscience, UK), 50 μM D (-)-2-amino-5-phosphonovaleric acid (APV, NMDAR-antagonist, Sigma-Aldrich, Israel), 200 μM (±)-a-Methyl-(4-carboxyphenyl)glycine (MCPG, mGluR-antagonist, Sigma-Aldrich, Israel), KT5720 (protein kinase A inhibitor, Tocris Bioscience, UK), GF109203x (protein kinase C inhibitor, Tocris Bioscience, UK).

Techniques: Activation Assay, Transmission Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

doi: 10.3389/fnmol.2017.00042

Figure Lengend Snippet: PAR2 induces LTD through the activation of TRPV4. (A) Application of TRPV4-agonist (2 μM RN1747) causes LTD. (B) Removal of the TRPV4-agonist (2 μM RN1747) following induction of LTD does not affect the stability of synaptic depression. (C) In presence of the TRPV4-antagonist (10 μM RN1734) the TRPV4-agonist is not able to induce synaptic depression. (D) In a two pathways experimental setting, low frequency stimulation (LFS, 1 Hz, 900 pulses) and TRPV4-agonist application induce similar levels of LTD. (E) LFS-induced LTD is not blocked by the TRPV4-antagonist. (F) Application of PAR2-agonist (10 μM AC55541) in presence of a TRPV4-antagonist (10 μM RN1734) blocks PAR2-induced LTD. (G) Application of TRPV4-agonist (2 μM RN1747) in presence of PAR2-antagonist (50 μM FSLLRY-NH 2 ) does not affect TRPV4-induced LTD. (H) Once PAR2-agonist mediated LTD is established, the TRPV4-agonist (2 μM RN1747) does not further de-potentiate a second pathway at adjusted response level (upward arrow). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section, n = 12 slices for each experiments, refer to text for statistics.

Article Snippet: The following compounds were used at the following concentrations: 10 μM AC55541 (PAR2-agonist, Tocris Bioscience, UK), 10 μM AC264613 (PAR2-agonist, Tocris Bioscience, UK), 50 μM FSLLRY-NH 2 (PAR2-antagonist, Sigma-Aldrich, Israel), 2 μM RN1747 (TRPV4-agonist, Tocris Bioscience, UK), 10 μM RN1734 (TRPV4-antagonist, Tocris Bioscience, UK), 10 μM RN9893 (TRPV4-antagonist, Tocris Bioscience, UK), 50 μM D (-)-2-amino-5-phosphonovaleric acid (APV, NMDAR-antagonist, Sigma-Aldrich, Israel), 200 μM (±)-a-Methyl-(4-carboxyphenyl)glycine (MCPG, mGluR-antagonist, Sigma-Aldrich, Israel), KT5720 (protein kinase A inhibitor, Tocris Bioscience, UK), GF109203x (protein kinase C inhibitor, Tocris Bioscience, UK).

Techniques: Activation Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

doi: 10.3389/fnmol.2017.00042

Figure Lengend Snippet: PAR2-mediated LTD is protein kinase A (PKA)-dependent. (A) PAR2-agonist (10 μM AC55541) in presence of a PKA-inhibitor (2 μM KT5720) KT5720 (2 μM) fails to induce LTD. (B) Application of a protein kinase C (PKC) inhibitor (2 μM GF109203x) does not affect the induction of PAR2-mediated LTD. Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section.

Article Snippet: The following compounds were used at the following concentrations: 10 μM AC55541 (PAR2-agonist, Tocris Bioscience, UK), 10 μM AC264613 (PAR2-agonist, Tocris Bioscience, UK), 50 μM FSLLRY-NH 2 (PAR2-antagonist, Sigma-Aldrich, Israel), 2 μM RN1747 (TRPV4-agonist, Tocris Bioscience, UK), 10 μM RN1734 (TRPV4-antagonist, Tocris Bioscience, UK), 10 μM RN9893 (TRPV4-antagonist, Tocris Bioscience, UK), 50 μM D (-)-2-amino-5-phosphonovaleric acid (APV, NMDAR-antagonist, Sigma-Aldrich, Israel), 200 μM (±)-a-Methyl-(4-carboxyphenyl)glycine (MCPG, mGluR-antagonist, Sigma-Aldrich, Israel), KT5720 (protein kinase A inhibitor, Tocris Bioscience, UK), GF109203x (protein kinase C inhibitor, Tocris Bioscience, UK).

Techniques:

Journal: Shock

Article Title: Inhibition of PAR-2 Attenuates Neuroinflammation and Improves Short-Term Neurocognitive Functions Via ERK1/2 Signaling Following Asphyxia-Induced Cardiac Arrest in Rats

doi: 10.1097/shk.0000000000001516

Figure Lengend Snippet: FIG. 3. Effect of FSLLRY-NH2 on neurocognitive functions after ACA. Effects of FSLLRY-NH2 on neurological functions were assessed by (A) NDS at 24 and 48 h, (B) number of seizures at 24 and 48 h, and (C) T-maze test at 7 days after ACA. ACA significantly worsened performance while treatment with FSLLRY- NH2 at a dose of 50 mg significantly improved neurological functions in all tests following ACA. FSLLRY-NH2 treatment at a dose of 100 mg was not effective in NDS at 24 h and T-maze test. AC55541 significantly deteriorated neurobehavioral outcome at all time points compared to the vehicle treated ACA group. Data are expressed as mean SD. n ¼ 6/group. ANOVA, Tukey. **P < 0.001 vs. Sham group, *P < 0.05 vs. Sham group, #P < 0.05 vs. ACA þ vehicle group. ACA indicates asphyxial cardiac arrest; d, days; h, hours.

Article Snippet: Selective PAR-2 activator AC55541 (30 mg/rat) and ERK1/2 inhibitor PD98059 (Santa Cruz Biotechnology, Dallas, Tex; 2 mL of 2 mmol/L) were used for intervention (25).

Techniques:

Journal: Shock

Article Title: Inhibition of PAR-2 Attenuates Neuroinflammation and Improves Short-Term Neurocognitive Functions Via ERK1/2 Signaling Following Asphyxia-Induced Cardiac Arrest in Rats

doi: 10.1097/shk.0000000000001516

Figure Lengend Snippet: FIG. 4. Effect of FSLLRY-NH2 on neuronal degeneration after ACA. Representative microphotographs (A) and quantitative analyses (B) of FJC-positive cells revealed significant neuronal degeneration in hippocampal CA1 region at 7 days after ACA. AC55541 further exacerbated the number of FJC-positive cells in rats subjected to ACA while FSLLRY-NH2 at doses of 50 mg and 150 mg markedly reduced hippocampal neuronal degeneration. Data are expressed as mean SD. n ¼ 6/group. ANOVA, Tukey. Scale bar ¼ 100 mm. **P < 0.001 vs. Sham group, ##P < 0.001 vs. ACA þ vehicle group, #P < 0.05 vs. ACA þ vehicle group. ACA indicates asphyxial cardiac arrest.

Article Snippet: Selective PAR-2 activator AC55541 (30 mg/rat) and ERK1/2 inhibitor PD98059 (Santa Cruz Biotechnology, Dallas, Tex; 2 mL of 2 mmol/L) were used for intervention (25).

Techniques:

Journal: Shock

Article Title: Inhibition of PAR-2 Attenuates Neuroinflammation and Improves Short-Term Neurocognitive Functions Via ERK1/2 Signaling Following Asphyxia-Induced Cardiac Arrest in Rats

doi: 10.1097/shk.0000000000001516

Figure Lengend Snippet: FIG. 5. Inhibition of ERK1/2 abolished the neuroinflammatory effect of PAR-2 activation at 24 h after ACA. Representative Western blot images (A) and quantitative analysis of PAR-2 (B), ERK1/2 (C), p-ERK1/2 (D), IL-6 (E), and TNF- a (F) in the brain revealed increased protein levels at 24 h following ACA except for ERK1/2 compared to the Sham group. Treatment with FSLLRY-NH2 significantly reduced p-ERK1/2 and proinflammatory cytokine levels compared to the ACA þ vehicle group. Further activation of PAR-2 with AC55541 only aggravated the neuroinflammatory response by increasing p-ERK1/2 expression. Potent ERK1/2 inhibitor PD98059 reversed the neuroinflammatory effect of AC55541. Neurologic outcome assessment with NDS at 24 h following ACA (G) revealed that the inhibition of PAR-2 significantly improved neurologic function while AC55541 alone significantly worsened performance compared to the ACA þ vehicle group. This detrimental effect of AC55541 on NDS was reversed by the ERK1/2 inhibitor, PD98059. Data are expressed as mean SD. n ¼ 6/group. ANOVA, Tukey. **P < 0.001 vs. Sham group, *P < 0.05 vs. Sham group, #P < 0.05 vs. ACA þvehicle group, &&P < 0.001 compared to ACA þ AC55541 group, &P < 0.05 compared to ACA þ AC55541 group. ACA indicates asphyxial cardiac arrest.

Article Snippet: Selective PAR-2 activator AC55541 (30 mg/rat) and ERK1/2 inhibitor PD98059 (Santa Cruz Biotechnology, Dallas, Tex; 2 mL of 2 mmol/L) were used for intervention (25).

Techniques: Inhibition, Activation Assay, Western Blot, Expressing