specific ep3 receptor agonists  (Ono Pharma)

Journal: Frontiers in Neural Circuits

Article Title: Prostaglandin E2 Exerts Biphasic Dose Response on the PreBötzinger Complex Respiratory-Related Rhythm

doi: 10.3389/fncir.2022.826497

Figure Lengend Snippet: Overview of prostanoid receptors and pharmacology.

Article Snippet: The following drugs were used: EP1 antagonist GW848867X (Cayman Chemicals, Ann Arbor, MI, United States, 50 nM), EP2 Antagonist PF-04418948 (Cayman Chemical, 500 nM), EP3 antagonist DG041 (Tocris, Bristol, United Kingdom, 40 nM), EP4 antagonist MK2894 (APExBIO, Houston, TX, United States, 100 nM), Prostaglandin E2 (Sigma-Aldrich, St. Louis, MO, United States, 1 nM-1 μM), Prostaglandin F receptor antagonist AL-8810 (Cayman Chemical, 1 μM), Prostaglandin D2 receptor antagonist Setipiprant (Cayman Chemical, 500 nM), Non-selective EP2/3/4 agonist 11-deoxy PGE1 (Cayman Chemical, 10 nM), EP2R agonist Butaprost (Cayman Chemical, 5 μM), EP3 receptor agonist Sulprostone (Cayman Chemical, 1 μM).

Techniques:

Journal: Frontiers in Neural Circuits

Article Title: Prostaglandin E2 Exerts Biphasic Dose Response on the PreBötzinger Complex Respiratory-Related Rhythm

doi: 10.3389/fncir.2022.826497

Figure Lengend Snippet: Integrated XII motor output in preBötC slices after exposure to prostanoid receptor agonists. (A) Examples of integrated hypoglossal motor output (XII) of slices exposed to EP2R agonist Butaprost (500 nM), EP3R agonist Sulprostone (1 μM) and non-selective prostanoid receptor agonist 11-deoxy PGE1 (10 nM) (150 s shown per condition). (B) Activating prostanoid receptors with selective agonists modulates burst period of XII motor output. Exposure to EP2R agonist Butaprost decreases burst period (–28% below control, p < 0.01), while EP3R agonist Sulprostone increases burst period (+21% above control, p < 0.0001). Non-selective prostanoid receptor agonist 11-deoxy PGE1 does not change the burst period. (C) EP2R agonist Butaprost decreases average burst amplitude (–35% below control, p < 0.001). (D) EP3R agonist Sulprostone decreases burst width (–9% below control, p < 0.01). (E,F) EP2R agonist Butaprost decreases XII burst area (–44% below control, p < 0.001) and the CV of the burst period (–0.07 below control, p < 0.05) (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: The following drugs were used: EP1 antagonist GW848867X (Cayman Chemicals, Ann Arbor, MI, United States, 50 nM), EP2 Antagonist PF-04418948 (Cayman Chemical, 500 nM), EP3 antagonist DG041 (Tocris, Bristol, United Kingdom, 40 nM), EP4 antagonist MK2894 (APExBIO, Houston, TX, United States, 100 nM), Prostaglandin E2 (Sigma-Aldrich, St. Louis, MO, United States, 1 nM-1 μM), Prostaglandin F receptor antagonist AL-8810 (Cayman Chemical, 1 μM), Prostaglandin D2 receptor antagonist Setipiprant (Cayman Chemical, 500 nM), Non-selective EP2/3/4 agonist 11-deoxy PGE1 (Cayman Chemical, 10 nM), EP2R agonist Butaprost (Cayman Chemical, 5 μM), EP3 receptor agonist Sulprostone (Cayman Chemical, 1 μM).

Techniques:

Journal: Frontiers in Immunology

Article Title: Myeloid-Derived Suppressor Cells Dampen Airway Inflammation Through Prostaglandin E2 Receptor 4

doi: 10.3389/fimmu.2021.695933

Figure Lengend Snippet: The effect of PGE2 and EP receptor agonists on MDSC generation in murine bone marrow cells. Bone marrow cells isolated from the hind legs of 6- to 8-week-old female BALB/c mice were cultured in the presence of PGE2 or EP receptor agonists, including EP2 agonist Butaprost, dual EP1/EP3 agonist Sulprostone and EP4 agonist L-902,688 (1 or 10 µM) or their respective vehicles in combination with GM-CSF (20 ng/ml) and IL-6 (20 ng/ml) for 3 days. (A) Gating strategy of MDSC subtyping by flow cytometry. CD11b + cells were first selected from live bone marrow cells before MDSCs were gated. (B) Representative MDSC gating of bone marrow cells cultured in either vehicle (left) or 10 µM agonist (right), where CD11b + Ly6G + Ly6C int and CD11b + Ly6G - Ly6C hi are defined as PMN- and M-MDSCs, respectively. (C) Quantitative analysis of both PMN- (left) and M-MDSCs (right) were performed. Data shown as the percentage of live cells. Data was obtained from 5-9 independent experiments, with bone marrow cells from each mouse divided into the different conditions (vehicle, 1 or 10 µM) of each experiment, including 2 technical replicates per condition. Results of individual experiments and mean ± SD are shown. Statistical analysis was performed with repeated measures one-way ANOVA followed by Tukey’s multiple comparisons test. * p < 0.05, *** p < 0.001 compared with vehicle, # p < 0.05 compared to 1 µM.

Article Snippet: The following compounds were tested: PGE2, EP2 receptor agonist Butaprost, EP1/EP3 receptor agonist Sulprostone and EP4 receptor agonist L-902,688 (all from Cayman Chemical, Ann Arbor, USA).

Techniques: Isolation, Cell Culture, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Myeloid-Derived Suppressor Cells Dampen Airway Inflammation Through Prostaglandin E2 Receptor 4

doi: 10.3389/fimmu.2021.695933

Figure Lengend Snippet: The effect of PGE2 and EP receptor agonists on the immunosuppressive activity of MDSCs. Bone marrow cells isolated from hind legs of 6- to 8-week-old female BALB/c mice were cultured in the presence of PGE2 or EP receptor agonists, including EP2 agonist Butaprost, dual EP1/EP3 agonist Sulprostone and EP4 agonist L-902,688 (all at 10 µM) or their respective vehicles, in combination with GM-CSF (20 ng/ml) and IL-6 (20 ng/ml). After 3 days of culture, both PMN- and M-MDSCs were isolated. CD4 + T cells were isolated from the spleens of 6- to 8-week-old female BALB/c mice. The MDSCs were then co-cultured with CFSE-labeled CD4 + T cells in the presence of anti-CD3/CD28. CFSE dilution was evaluated by flow cytometry after 3 days. (A) Gating strategy to assess CD4 + T-cell proliferation by flow cytometry. CFSE + CD4 + cells were first selected from viable cells after which the percentages of CD4 + T-cell proliferation were analysed (histogram). (B) Representative histograms showing the CFSE dilutions in CD4 + T cells that were co-cultured together with PMN- or M-MDSCs (pre-incubated with 10 µM L-902,688 or vehicle) at different ratios. (C) Analysis of the immunosuppressive activity of PMN- (left) and M-MDSCs (right). Data represents the normalised percentages of proliferating CD4 + T cells (positive control, without MDSCs, set to 100%). Data were pooled from 3-4 experiments with 2 technical replicates per experiment. Results are presented as mean ± SD. Statistical analysis was performed comparing each MDSC: CD4 + T cell ratio with unpaired two-tailed Student’s t-tests. * p < 0.05, *** p < 0.001 compared with vehicle.

Article Snippet: The following compounds were tested: PGE2, EP2 receptor agonist Butaprost, EP1/EP3 receptor agonist Sulprostone and EP4 receptor agonist L-902,688 (all from Cayman Chemical, Ann Arbor, USA).

Techniques: Activity Assay, Isolation, Cell Culture, Labeling, Flow Cytometry, Incubation, Positive Control, Two Tailed Test

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Central EP3 receptors mediate salt sensitive hypertension and immune activation

doi: 10.1161/HYPERTENSIONAHA.119.13850

Figure Lengend Snippet: Effects of EP3 deficiency on L-NAME/high salt induced hypertension and autonomic function. Wild type and EP3−/− mice received 0.5 mg/mL L-NAME (LN) in the drinking water, followed by a 2-week washout (WO) and subsequently a 4% high-salt diet. Systolic blood pressure (BP), diastolic BP and heart rate (panels A) were measured by telemetry. Power spectral analysis of blood pressure and heart rate variabilities at night time are shown in B and C. Spot urine was collected at the end of high salt treatment, and urinary norepinephrine (NE) was determined by HPLC as shown in D. Blood pressure and heart rate data were analyzed by area under the curve followed with t test, data from the power spectral analysis and urinary norepinephrine were analyzed with 2-way ANOVA and Bonferroni post-hoc multiple comparisons. n=5 to 9 in each group. *P<0.05, **P<0.01.

Article Snippet: PGE 2 , EP1 receptor antagonist SC-51322, EP3 receptor antagonist DG-041, EP1 or EP3 receptor agonists 17-phenyl-trinor-PGE2 (Cayman Chemical, Ann Arbor, MI), sulprostone (Cayman Chemical, Ann Arbor, MI), MB-28767 (a gift from Dr. M.P.L.

Techniques:

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Central EP3 receptors mediate salt sensitive hypertension and immune activation

doi: 10.1161/HYPERTENSIONAHA.119.13850

Figure Lengend Snippet: Effects of EP3 deficiency on renal oxidative stress and renal fibrosis in response to L-NAME/high salt (LN/HS) induced hypertension. DHE fluorescence was visualized at 530 nm to 560 nm and relative fluorescence intensity was quantified in (A) and (B). Collagen was identified using Masson’s trichrome blue stain and quantified in (C) and (D). Data were analyzed using 2-way ANOVA and post-hoc multiple comparisons. *P<0.05, **P<0.01, n= 4 to 8 in each group. Black lines denote 100 μm.

Article Snippet: PGE 2 , EP1 receptor antagonist SC-51322, EP3 receptor antagonist DG-041, EP1 or EP3 receptor agonists 17-phenyl-trinor-PGE2 (Cayman Chemical, Ann Arbor, MI), sulprostone (Cayman Chemical, Ann Arbor, MI), MB-28767 (a gift from Dr. M.P.L.

Techniques: Fluorescence, Staining

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Central EP3 receptors mediate salt sensitive hypertension and immune activation

doi: 10.1161/HYPERTENSIONAHA.119.13850

Figure Lengend Snippet: Effects of EP3 deficiency on renal leukocyte and T-cell infiltration. Mice underwent L-NAME/high salt (LN/HS) protocol as in figure 1. Flow cytometry gating strategy of kidneys are shown in (A). Live singlet cells were gated for total leukocytes (CD45+), monocytes/macrophages (F4/80+), total T cells (CD3+), CD4+ and CD8+ T cells. Representative intracellular staining for IFN-γ and IL-17A in CD3+ T cells from wild type and EP3−/− mice, as well as respective fluorescence-minus-one (FMO) controls are shown in (B). Mean data are shown in (C). Data were analyzed using 2-way ANOVA followed by Bonferroni post-hoc test, n=5 to 12 in each group. *P<0.05, **P<0.01.

Article Snippet: PGE 2 , EP1 receptor antagonist SC-51322, EP3 receptor antagonist DG-041, EP1 or EP3 receptor agonists 17-phenyl-trinor-PGE2 (Cayman Chemical, Ann Arbor, MI), sulprostone (Cayman Chemical, Ann Arbor, MI), MB-28767 (a gift from Dr. M.P.L.

Techniques: Flow Cytometry, Staining, Fluorescence

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Central EP3 receptors mediate salt sensitive hypertension and immune activation

doi: 10.1161/HYPERTENSIONAHA.119.13850

Figure Lengend Snippet: Effects of EP3 deficiency on dendritic cell activation and isoLG adduct formation in L-NAME/high salt (LN/HS) induced hypertension. DCs were gated after exclusion of macrophages, and expression of costimulatory molecules CD86, and intracellular isoLG-protein adducts were measured. Representative results are shown in (A), and mean data are shown in (B) and (C). Fluorescence minus one controls (FMO) were shown in black dashed lines. Data were analyzed using 2-way ANOVA. *P<0.05, n= 4 to 6 in each group.

Article Snippet: PGE 2 , EP1 receptor antagonist SC-51322, EP3 receptor antagonist DG-041, EP1 or EP3 receptor agonists 17-phenyl-trinor-PGE2 (Cayman Chemical, Ann Arbor, MI), sulprostone (Cayman Chemical, Ann Arbor, MI), MB-28767 (a gift from Dr. M.P.L.

Techniques: Activation Assay, Expressing, Fluorescence

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Central EP3 receptors mediate salt sensitive hypertension and immune activation

doi: 10.1161/HYPERTENSIONAHA.119.13850

Figure Lengend Snippet: Role of central EP3 receptors in L-NAME/high salt induced hypertension. Punch biopsies of the subfornical organ (SFO), organum vasculosum laminae terminalis (OVLT), paraventricular nucleus (PVN) and cortex were collected from mice after normal salt (NS), L-NAME only (LN), and L-NAME/high salt (LN/HS). Gene expression of COX-1, COX-2, and PGE2 receptors (EP1-EP4) was determined by real time PCR shown in (A). Beta-actin was used as control gene to calculate ΔCT, n=3 to 9 in each group. Knockdown EP3 receptor was accomplished by ICV injection of 107 lentivirus vectors encoding shRNAs targeting EP3 gene or scrambled control sequences. After recovery from surgery, mice were subjected to L-NAME/high salt protocol, and systolic blood pressure (BP), diastolic BP and heart rate were recorded using telemetry shown in (B). Data were analyzed by area under the curve followed with t test. Power spectra analyses (PSA) of blood pressure and heart rate variability (BPV and HRV) are shown in panel (C). After animals were sacrificed, EP3 receptor mRNA levels in OVLT, SFO, PVN, and cortex were determined by real time PCR shown in (D). Data were analyzed by multiple t tests, *P<0.05, **P<0.01, n=7 in both groups.

Article Snippet: PGE 2 , EP1 receptor antagonist SC-51322, EP3 receptor antagonist DG-041, EP1 or EP3 receptor agonists 17-phenyl-trinor-PGE2 (Cayman Chemical, Ann Arbor, MI), sulprostone (Cayman Chemical, Ann Arbor, MI), MB-28767 (a gift from Dr. M.P.L.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control, Knockdown, Injection

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Central EP3 receptors mediate salt sensitive hypertension and immune activation

doi: 10.1161/HYPERTENSIONAHA.119.13850

Figure Lengend Snippet: Role of central EP3 receptors in L-NAME/high salt induced renal inflammation. IsoLG adducts in splenic DCs were quantified by flow cytometry shown in (A). Total leukocytes (CD45+), total T lymphocytes (CD3+), monocytes/macrophages (F4/80+), and CD4+ and CD8+ T cell subsets in the kidney were quantified by flow cytometry shown in from panel B to E. Data were analyzed by student t tests. P<0.05, **P<0.01, n=7 in both groups.

Article Snippet: PGE 2 , EP1 receptor antagonist SC-51322, EP3 receptor antagonist DG-041, EP1 or EP3 receptor agonists 17-phenyl-trinor-PGE2 (Cayman Chemical, Ann Arbor, MI), sulprostone (Cayman Chemical, Ann Arbor, MI), MB-28767 (a gift from Dr. M.P.L.

Techniques: Flow Cytometry