ox1r antagonist  (Tocris)

Journal: Regenerative Therapy

Article Title: Orexin-A increases the differentiation of human olfactory sensory neurons through orexin receptor type 1

doi: 10.1016/j.reth.2024.10.014

Figure Lengend Snippet: Immunofluorescence of orexin receptors and dose-response analyses of orexin-A. (a) Immunofluorescence images of OSNs stained with OX1R and OX2R (green). (b) Schematic showing the proposed mechanism of orexin-mediated differentiation of OSNs and dose-response analyses of suvorexant. Western blot densitometric analysis of (c) ASCL1, (d) βIII Tubulin, and (e) OMP, respectively. (p > 0.05, compared to controls, n = 5).

Article Snippet: The orexin receptor type 1 (OX1R) antagonist, SB-674042 (HY-10898, MedChem Express, NJ, USA), and orexin receptor type 2 (OX2R) antagonist, TCS-OX2-29 (HY-100452, MedChem Express) were diluted in the dimethyl sulfoxide, DMSO with 1 μg/mL and 10 μg/mL, respectively.

Techniques: Immunofluorescence, Staining, Western Blot

Journal: Regenerative Therapy

Article Title: Orexin-A increases the differentiation of human olfactory sensory neurons through orexin receptor type 1

doi: 10.1016/j.reth.2024.10.014

Figure Lengend Snippet: The effect of OX1R antagonists (SB674042) and OX2R antagonists (TCS-OX2-29) on the differentiation of OSN. The expression of various markers was assessed through western blots (a and g) and subsequently analyzed densitometrically for ADCY3, Golf, ASCL1, βIII tubulin, and OMP (b-f and h-l). (Asterisks indicate p < 0.05, compared to controls, n = 5–6).

Article Snippet: The orexin receptor type 1 (OX1R) antagonist, SB-674042 (HY-10898, MedChem Express, NJ, USA), and orexin receptor type 2 (OX2R) antagonist, TCS-OX2-29 (HY-100452, MedChem Express) were diluted in the dimethyl sulfoxide, DMSO with 1 μg/mL and 10 μg/mL, respectively.

Techniques: Expressing, Western Blot

Journal: Translational psychiatry

Article Title: Lateral hypothalamus orexinergic projection to the medial prefrontal cortex modulates chronic stress-induced anhedonia but not anxiety and despair.

doi: 10.1038/s41398-024-02860-9

Figure Lengend Snippet: Fig. 7 Blocking OX1R or OX2R in the mPFC alleviated UCMS-induced anhedonia, but not anxiety and despair. A Experimental timeline of microinjection blocking the LHOrx-mPFC pathway. B Left: Schematic of microinjection. Right: Representative image of tube in the mPFC. Blocking either OX1R or OX2R did not prevent anxiety-like (C time in center of the OFT, D time in open arms of the EPM, E Latency to feed in the NSFT), despair-like phenotypes (F immobility time in the FST, G Immobility time in the TST). H In the UCMS mice, blocking OX1R or OX2R increased the grooming time in the SST. I Blocking OX1R or OX2R increased volume consumed in the SPT after UCMS. n (Ctrl + DMSO) = 11, n

Article Snippet: The Orx 1 receptor (Ox1R) antagonist SB334867 (SB, 25 mM, #1455, Tocris Bioscience, Bristol, UK) and Orx 2 receptor (Ox2R) antagonist TCS-OX2-29 (TCS, 33.3 μg/μL, #1457, Tocris Bioscience) dissolved in dimethyl sulfoxide (DMSO, #196055, MP Biomedicals, California, USA) and saline were injected to the mPFC, respectively [22].

Techniques: Blocking Assay, Microinjection

Journal: iScience

Article Title: An orexin-receptor-2-mediated heart-brain axis in cardiac pain

doi: 10.1016/j.isci.2024.109067

Figure Lengend Snippet: Epicardial application of OX2R agonists attenuates c-Fos activity in PVN (A) c-Fos staining in PVN 90 min after epicardial application of saline and 10 nmol/kg OB-Ala. (B) c-Fos staining in PVN 90 min after i.c.v. injection of saline and 10 nmol/kg OB-Ala. (C) Quantification of c-Fos-positive neuron numbers in PVN 90 min after epicardial application and i.c.v. injection of saline and OB-Ala at 1 nmol/kg, 10 nmol/kg, and 100 nmol/kg. (D) c-Fos staining in ARC 90 min after epicardial application of saline and 10 nmol/kg OB-Ala. (E) c-Fos staining in ARC 90 min after i.c.v. injection of saline and 10 nmol/kg OB-Ala. (F) Quantification of c-Fos-positive neuron numbers in ARC 90 min after epicardial application and i.c.v. injection of saline and OB-Ala at 1 nmol/kg, 10 nmol/kg, and 100 nmol/kg. (G) c-Fos staining in PVN 90 min after i.c.v. injection of 10 nmol/kg OA. (H) c-Fos staining in PVN 90 min after epicardial application of OA, OA together with OX2R antagonist or OX1R antagonist. (I) Quantification of (G) and (H). (J) c-Fos staining in ARC 90 min after i.c.v. injection of 10 nmol/kg OA. (K) c-Fos staining in PVN 90 min after epicardial application of OA, OA together with OX2R antagonist or OX1R antagonist. (L) Quantification of (J) and (K). N = 6–8 mice per group. Independent experiments were repeated two times. Data are presented as means ± SEM. Significant effect of each dosage by one-way ANOVA. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Scale bar: 100 μm. PVN, paraventricular nucleus; ARC, arcuate nucleus; OA, orexin A; OX1R, orexin receptor 1; OX2R, orexin receptor 2.

Article Snippet: 10μM OX1R specific antagonist SB334867 (MCE, HY-10895A) or OX2R specific antagonist EMPA (Sigma, SML0864) were added 10min before OA and capsaicin.

Techniques: Activity Assay, Staining, Saline, Injection

Journal: iScience

Article Title: An orexin-receptor-2-mediated heart-brain axis in cardiac pain

doi: 10.1016/j.isci.2024.109067

Figure Lengend Snippet: OX2R agonist blocks capsaicin-induced Ca 2+ flux in DRG (A) Fluo-8 indicated calcium images of cultured DGR neurons at 0 s and 30 s after Veh, 10 μM capsaicin, capsaicin + OB-Ala, capsaicin + OA, capsaicin + OA + OX2R antagonist, and capsaicin + OA + OX1R antagonist. (B) Quantification of calcium fluorescent change rate peak (about 30 s) in (A). 17/42/47/34/81/61 cells from five isolations were recorded for (A). Significant effect of each treatment by one-way ANOVA. Data are presented as means ± SEM. ∗∗∗∗p < 0.0001. Scale bar: 20 μm.

Article Snippet: 10μM OX1R specific antagonist SB334867 (MCE, HY-10895A) or OX2R specific antagonist EMPA (Sigma, SML0864) were added 10min before OA and capsaicin.

Techniques: Cell Culture

Journal: iScience

Article Title: An orexin-receptor-2-mediated heart-brain axis in cardiac pain

doi: 10.1016/j.isci.2024.109067

Figure Lengend Snippet: OX2R agonist inhibits c-Fos activity in the nucleus involved in capsaicin-mediated cardiac sympathetic afferent reflex c-Fos activity in PVN (A and B), NTS (C and D), and PAG (E and F) after epicardial application of saline, capsaicin, capsaicin + OB-Ala, and capsaicin + lidocaine. c-Fos activity in PVN (G and H), NTS (I and J), and PAG (K and L) after epicardial application of capsaicin + OA, capsaicin + OA + OX2R antagonist, and capsaicin OA + OX1R antagonist. N = 6–10 mice per group. Independent experiments were repeated at least two times. Significant effect of each treatment by one-way ANOVA. Data are presented as means ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Scale bar: 100 μm in all panels.

Article Snippet: 10μM OX1R specific antagonist SB334867 (MCE, HY-10895A) or OX2R specific antagonist EMPA (Sigma, SML0864) were added 10min before OA and capsaicin.

Techniques: Activity Assay, Saline

Journal: iScience

Article Title: An orexin-receptor-2-mediated heart-brain axis in cardiac pain

doi: 10.1016/j.isci.2024.109067

Figure Lengend Snippet:

Article Snippet: 10μM OX1R specific antagonist SB334867 (MCE, HY-10895A) or OX2R specific antagonist EMPA (Sigma, SML0864) were added 10min before OA and capsaicin.

Techniques: Recombinant, RNAscope, Calcium Assay, Enzyme-linked Immunosorbent Assay, Software