Journal: FEBS Open Bio

Article Title: The photoswitchable cannabinoid azo ‐ HU308 enables optical control of Ca 2+ dynamics in INS ‐1 β‐cells via off‐target effects on TRPC channels

doi: 10.1002/2211-5463.70146

Figure Lengend Snippet: Azo‐HU308's photoswitching effect in INS‐1 cells is not mediated by CB2R or G protein signaling. INS‐1 cells were transfected with R‐GECO and Ca 2+ levels recorded by confocal microscopy. Azo ‐HU308 (20 μ m ) and 375 nm light were applied in conjunction with various signaling antagonists. Shown are: (A) Summary bar graph showing that the % Ca 2+ effect for azo ‐HU308 photoswitching in the presence of CB2R inhibitors AM630 (20 μ m ; N = 151, T = 3), SR144528 (20 μ m ; N = 159, T = 3), and JWH133 (32 μ m ; N = 166, T = 3), which failed to block azo ‐HU308's effect. Similar results were observed with the CB1R inhibitor Rimanobant (2 μ m ; N = 103, T = 2), GPR55 inhibitor CID16020047 (20 μ m ; N = 104, T = 2), PLC inhibitor U73122 (10 μ m ; N = 89, T = 2), or the IP 3 R inhibitor Xestospongin C (1 μ m ; N = 152, T = 3). Inhibitors for G protein signaling, including G s ‐protein inhibitor NF449 (10 μ m ; N = 173, T = 3), G i/o ‐protein inhibitor pertussis toxin (100 ng·mL −1 ; N = 135, T = 4), and G q/11 protein inhibitor YM524890 (10 μ m ; N = 120, T = 3) also failed to block azo ‐HU308 photoswitching. (B) Averaged Ca 2+ traces for CB2R inhibitors AM630 (red), SR144528 (blue), and JWH133 (black). (C) Average traces for U73122 (green) and Xestospongin C (orange). (D) Average Ca 2+ traces for G protein inhibitors: G s ‐inhibitor NF449 (cyan), G i/o ‐inhibitor pertussis toxin (magenta) and G q/11 ‐inhibitor YM254890 (yellow). Error bars = mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant. Student's t ‐test.

Article Snippet: HU308 (Cayman Chemical, Ann Arbor, MI, USA; #90086), AM630 (Cayman Chemical, #1006974), SR 144528 (Cayman Chemical, #9000491), JWH 133 (Cayman Chemical, #10005428), Rimonabant hydrochloride (Sigma‐Aldrich, St. Louis, MO, USA; #SML0800), CID16020046 (Tocris, Bristol, UK; #4959), Xestospongin C (Cayman Chemical, #64950), U73122 (Tocris, #1268), YM254890 (Tocris, #7352), Pertussis Toxin (Cayman Chemical, #19546), NF449 (Cayman Chemical, #13324), 2‐APB (Tocris, #1224), Capsazepine (Cayman Chemical, #10007518), SAR7334 hydrochloride (Cayman Chemical, #28292), SKF 96365 hydrochloride (Cayman Chemical, #10009312), ML‐204 (Cayman Chemical, #15626), YM‐58483 (Cayman Chemical, #13246), and Pyr 10 (Tocris, #6941) were obtained from commercial suppliers.

Techniques: Transfection, Confocal Microscopy, Blocking Assay

Journal: Cells

Article Title: Intranasal Treatment with Cannabinoid 2 Receptor Agonist HU-308 Ameliorates Cold Sensitivity in Mice with Traumatic Trigeminal Neuropathic Pain.

doi: 10.3390/cells13231943

Figure Lengend Snippet: Figure 4. Comparative analysis of the effects of intranasal and oral administration of CB2 agonist on cold hypersensitivity in IONC mice. The area under the curve was determined from the acetone test data following either intranasal (i.n.) or oral (p.o.) repeated administration of HU-308 (HU, 30 nmole, 10 µL) and compared. Individual data and mean ± SEM are shown. N = 5; * p < 0.05, ** p < 0.01 (two-way ANOVA followed by Tukey’s multiple comparisons test).

Article Snippet: Regardless of the administration methods, 30 nmol of CB2 agonist HU-308 (Cat. #: 90086; Cayman Chemical, Ann Arbor, MI, USA) and 100 nmol of CB2 antagonist SR 144528 (Cat. #: 9000491; Cayman Chemical, Ann Arbor, MI, USA) were used in this study.

Techniques: