Journal: Annals of Dermatology

Article Title: Selective Cannabinoid Receptor-1 Agonists Regulate Mast Cell Activation in an Oxazolone-Induced Atopic Dermatitis Model

doi: 10.5021/ad.2016.28.1.22

Figure Lengend Snippet: Newly synthesized cannabinoids and the cAMP assay screening for cannabinoid receptor modulators. (A) The structure of α-oleoyl oleoylamine ethanolamine (α-OOE) and α-oleoyl oleoylamine serinol (α-OOS). (B, C) Synthetic compounds were screened in stable CHO cell lines overexpressing cannabinoid receptor-1 (CB1R) or CB2R. Pre-treatment with CBR agonists significantly diminished the cytosolic levels of cAMP induced by 10 µM forskolin (B). N-arachidonoylethanolamine (AEA) and Hu308 were used as controls for CB1R and CB2R, respectively. All data are presented as the mean±standard error, t -test and analysis of variance. * p <0.01.

Article Snippet: To establish stable cell lines expressing CBR, CHO-K1 cells were transfected with human CB1R (Cat. RC210397) and CB2R (Cat. SC118984) cDNA constructs (Origene, Rockville, MD, USA); the following day, media were replaced with F12 standard medium containing G418 (100 μg/ml) to select for stable clones.

Techniques: Synthesized, cAMP Assay

Journal: Annals of Dermatology

Article Title: Selective Cannabinoid Receptor-1 Agonists Regulate Mast Cell Activation in an Oxazolone-Induced Atopic Dermatitis Model

doi: 10.5021/ad.2016.28.1.22

Figure Lengend Snippet: Effects of cannabinoid receptor-1 (CB1R) agonists on the degranulation of rat basophilic leukemia cell line (RBL-2H3) mast cells. (A, B) The inhibitory effects of α-oleoyl oleoylamine ethanolamine (α-OOE), α-oleoyl oleoylamine serinol (α-OOS), and N-arachidonoylethanolamine (AEA) were determined on ionomycin (2 µM)-induced histamine (A) or β-hexosaminidase release (B) from rat mast cells. (C, D) RBL-2H3 cells sensitized with anti-dinitropheno-immunoglobulin E (anti-DNP-IgE) (0.5 µg/ml) for 12 hr were pretreated with the CB1R agonists. Histamine (C) or β-hexosaminidase release (D) from mast cells was measured. (E) Bone marrow derived-cultured mast cells from mouse bone marrow was sensitized with anti-DNP-IgE for 12 hr and histamine release from cells was measured. All data are presented as the mean±standard error, t-test and analysis of variance. NON: non-stimulated, CON: control (stimulated), IgE: Immunoglobulin E, DNP: dinitrophenol, BSA: bovine serum albumin. * p <0.01 and ** p <0.05.

Article Snippet: To establish stable cell lines expressing CBR, CHO-K1 cells were transfected with human CB1R (Cat. RC210397) and CB2R (Cat. SC118984) cDNA constructs (Origene, Rockville, MD, USA); the following day, media were replaced with F12 standard medium containing G418 (100 μg/ml) to select for stable clones.

Techniques: Derivative Assay, Cell Culture, Control

Journal: Annals of Dermatology

Article Title: Selective Cannabinoid Receptor-1 Agonists Regulate Mast Cell Activation in an Oxazolone-Induced Atopic Dermatitis Model

doi: 10.5021/ad.2016.28.1.22

Figure Lengend Snippet: Inhibition of proliferation of rat basophilic leukemia cell line (RBL-2H3) cells by cannabinoid receptor-1 (CB1R) agonists. RBL-2H3 cells were untreated or treated with the indicated concentrations of CB1R agonists for 24 hr (A) or with 25 µM agonists for the indicated time periods (B). (C) RBL-2H3 cell proliferation was analyzed by counting cell numbers following CB1R agonist treatment for 48 hr. α-OOE: α-oleoyl oleoylamine ethanolamine, α-OOS: α-oleoyl oleoylamine serinol, AEA: N-arachidonoylethanolamine. Significant difference ( * p <0.01 in Student's t -test) as compared with control samples.

Article Snippet: To establish stable cell lines expressing CBR, CHO-K1 cells were transfected with human CB1R (Cat. RC210397) and CB2R (Cat. SC118984) cDNA constructs (Origene, Rockville, MD, USA); the following day, media were replaced with F12 standard medium containing G418 (100 μg/ml) to select for stable clones.

Techniques: Inhibition, Control

Journal: Annals of Dermatology

Article Title: Selective Cannabinoid Receptor-1 Agonists Regulate Mast Cell Activation in an Oxazolone-Induced Atopic Dermatitis Model

doi: 10.5021/ad.2016.28.1.22

Figure Lengend Snippet: Effects of cannabinoid receptor-1 agonists in oxazolone-induced atopic dermatitis mice models. (A) Histology of dorsal skin lesions. The dorsal skin of each mouse was removed and fixed for toluidine blue staining (×400). Images are representative of five mice. (B) Change of skin fold thickness in oxazolone model. α-oleoyl oleoylamine ethanolamine (α-OOE), α-oleoyl oleoylamine serinol (α-OOS), and N-arachidonoylethanolamine (AEA) treatment prevented the increase of skin fold thickness. (C) Quantitative histomorphometry of the number of mast cells. Mast cells were counted in the toluidine blue-stained slide. (D) Total blood histamine levels in serum. The histamine concentration of each mouse was measured according to a previously described method. Each point represents the mean±standard error. Data are from five to six mice. VEH: vehicle, DEXA: dexamethasone. * p <0.01, ** p <0.05 significantly different from the vehicle group (Student's t-test).

Article Snippet: To establish stable cell lines expressing CBR, CHO-K1 cells were transfected with human CB1R (Cat. RC210397) and CB2R (Cat. SC118984) cDNA constructs (Origene, Rockville, MD, USA); the following day, media were replaced with F12 standard medium containing G418 (100 μg/ml) to select for stable clones.

Techniques: Staining, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Cannabinoid Receptor Type 1 Regulates Drug Reward Behavior via Glutamate Decarboxylase 67 Transcription

doi: 10.3390/ijms221910486

Figure Lengend Snippet: Effects of CB1 knockdown on GAD67 expression in primary cultured neurons and METH-induced CPP. ( A ) Vehicle group was treated with vehicle (5% DMSO, 5% tween-20, 90% PBS) as the same amount of JWH-210 for 24 h. JWH-210 group was treated with 0.1 μM JWH-210 for 24 h. Mock + JWH-210 and CB1 overexpression (OE) + JWH-210 groups were treated with mock or CB1 ORF lentiviral particles (5 MOI) for 24 h before treatment with vehicle or 0.1 μM JWH-210 for 24 h. Immunostaining of CB1, Myc, and GAD67 was performed with specific antibodies (Scale bar: 200 μm). ( B ) Cultured neurons were treated with control or CB1 shRNA lentiviral particles. mRNA levels of ( C ) CB1 and ( D ) GAD67 were confirmed by using qPCR with each specific primer and normalized to the relative amplification of GAPDH. Data are expressed as the mean ± S.E. ( n = 6; Student’s t -test; * p < 0.05 and ** p < 0.01 vs. vehicle group). ( E ) After CB1 knockdown, 0.3 mg/kg METH-induced CPP scores (s) was measured by CPP test. Data are expressed as the mean ± S.E. ( n = 8) and were analyzed using two-way ANOVA followed by Bonferroni post-hoc t -test (* p < 0.05 and ** p < 0.01 vs. each saline group; # ## p < 0.001 vs. Control shRNA/METH group). OE: overexpression. TUJ1: beta-tubulin III. METH: methamphetamine.

Article Snippet: CB1 human open reading frame (ORF) clone lentiviral particles (c-Myc tagged) were obtained from Origene (Rockville, MD, USA) for CB1 overexpression.

Techniques: Expressing, Cell Culture, Over Expression, Immunostaining, shRNA, Amplification