Journal: PLoS ONE

Article Title: Chronic marijuana usage by human pancreas donors is associated with impaired islet function

doi: 10.1371/journal.pone.0258434

Figure Lengend Snippet: (A) Immunofluorescent staining of human pancreatic tissue from individuals with a history of non-use and chronic marijuana use. The sections were stained for CB1R (red) and insulin (green). Details of the staining are shown in the enlarged images. Two islet images are presented from non-users and marijuana-users. Confocal images were taken using Zeiss LSM 700. Scale bars, 50 μm. (B) Quantitative analysis of CB1R expression. The CB1R intensity per islet area was compared between the two donor groups. Samples from five non-user donors and from six marijuana-user donors were counted for the analysis. *** p<0.001. The values were expressed as mean ± SEM. Each dot in the bar graph represents calculated value from an islet. (C) Immunofluorescent staining of human pancreatic tissue from individuals with a history of chronic marijuana use and non-use. The sections were stained for CB1R (red), glucagon (green), and somatostatin (green). Details of the staining are shown in the enlarged images. Images are presented from samples from four non-users and four marijuana-users. Confocal images were taken using a Zeiss LSM 700 microscope. Scale bars, 50 μm.

Article Snippet: For pancreatic tissue (six chronic marijuana-users and five non-users), sections were incubated over night at 4°C with primary antibodies including rabbit anti-CB1R, which targets the human extracellular N-terminus of CB1R (1:200 dilution, Alomone Labs, Israel, Cat# ACR-001), guinea pig anti-insulin (1:400 dilution, Abcam, Cat # ab195956), mouse anti-glucagon (1:2,000 dilution, Sigma), rat anti-somatostatin (1:50 dilution, BioRad).

Techniques: Staining, Expressing, Microscopy

Journal: bioRxiv

Article Title: Tetrahydrocannabinol exposure to postejaculatory sperm compromises sperm structure, function, the epigenome, and early embryo development

doi: 10.64898/2026.03.23.713385

Figure Lengend Snippet: A) Immunocytochemistry images of bovine sperm, human placenta, bovine placenta, and mouse brain depicting nuclear staining with DAPI (blue), CB1 (red), and merged images. FITC-PSA fluorescence can be observed in the merged image of sperm (green), for identification of the sperm acrosome. B) Western blot image of bovine sperm and equine pituitary identifying a single and specific band at the expected 75 kD value. Scale bar = 50 μM.

Article Snippet: Primary antibody incubation for detection of CB1 include the addition of 50 μL of Anti-CB1(proteintech ® Cat No: 17978-1-AP) at a dilution of 1:50 in BB for 15 h at 4 ° C. Slides were then washed and 100μL of secondary antibody (Donkey-AntiRabbit Abcam Cat No: ab6799) was added at a dilution of 1:100 (in BB) for 1 h in the dark followed by washing.

Techniques: Immunocytochemistry, Staining, Fluorescence, Western Blot

Journal: bioRxiv

Article Title: Tetrahydrocannabinol exposure to postejaculatory sperm compromises sperm structure, function, the epigenome, and early embryo development

doi: 10.64898/2026.03.23.713385

Figure Lengend Snippet: Localization of CB1 in non-capacitated sperm (upper panel), to the post-acrosomal sheath as compared to loss of abundance when sperm are held in capacitating conditions (middle panel) as indicted by tyrosine phosphorylation staining (YPO 3-, yellow). Sperm exposed to THC under non-capacitating conditions exhibit decreased YPO 3 , CB1 and acrosome detection (green) (lower panel). Scale bar = 50 μm.

Article Snippet: Primary antibody incubation for detection of CB1 include the addition of 50 μL of Anti-CB1(proteintech ® Cat No: 17978-1-AP) at a dilution of 1:50 in BB for 15 h at 4 ° C. Slides were then washed and 100μL of secondary antibody (Donkey-AntiRabbit Abcam Cat No: ab6799) was added at a dilution of 1:100 (in BB) for 1 h in the dark followed by washing.

Techniques: Phospho-proteomics, Staining

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

Journal: The Journal of Neuroscience

Article Title: Active Zone Proteins RIM1αβ Are Required for Normal Corticostriatal Transmission and Action Control

doi: 10.1523/JNEUROSCI.1940-18.2018

Figure Lengend Snippet: Gross brain anatomy is unaltered in conditional RIM1 KO mice. A–I, Neurofilament, CB1R, mu opioid receptor (MOPR), parvalbumin (PV), and/or DAPI staining of striatum (A–D), motor cortex (E,F) and hippocampus (G–I) of RIM1fl/fl and Emx1::Cre;RIM1−/− mice. Scale bars: A–D, G–I, 400 μm; E, F, 200 μm.

Article Snippet: Mesoscale cortical and hippocampal structure was similarly preserved in these mice, as indicated by labeling with DAPI and immunostaining for neurofilament, parvalbumin, and CB1R ( D–I ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2. caption a7 Gross brain anatomy is unaltered in conditional RIM1 KO mice.

Techniques: Staining

Journal: The Journal of Neuroscience

Article Title: Active Zone Proteins RIM1αβ Are Required for Normal Corticostriatal Transmission and Action Control

doi: 10.1523/JNEUROSCI.1940-18.2018

Figure Lengend Snippet: Intact short- and long-term depression of excitatory synaptic transmission in conditional RIM1 KO mice. A, DSE in DLS of RIM1fl/fl and Emx1::Cre;RIM1−/− mice. Arrow indicates 10 s period of postsynaptic depolarization to 0 mV. B, DHPG-induced LTD of EPSCs in DLS of RIM1fl/fl and Emx1::Cre;RIM1−/− mice. C, Effects of CB1R antagonist AM251 preincubation on DHPG-mediated LTD of EPSCs in DLS of RIM1fl/fl and Emx1::Cre;RIM1−/− mice. D, LTD of EPSCs in DLS of RIM1fl/fl and Emx1::Cre;RIM1−/− mice induced by bath application of mGlu2/3 receptor agonist, LY379268. Calibration: 10 ms, 100 pA.

Article Snippet: Mesoscale cortical and hippocampal structure was similarly preserved in these mice, as indicated by labeling with DAPI and immunostaining for neurofilament, parvalbumin, and CB1R ( D–I ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2. caption a7 Gross brain anatomy is unaltered in conditional RIM1 KO mice.

Techniques: Transmission Assay

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: Structure of representative CB1 allosteric modulators

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques:

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: PAM activity of fluoro-analogs of GAT211

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques: Activity Assay

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: PAM activity of aza-analogs of GAT211

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques: Activity Assay, Solubility

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: Effect of 6r, 9j, and 6s on the binding of a) 1 nM [3H]SR141716A or b) [3H]CP55,940 to membranes obtained from CHO cells stably-expressing human CB1R; or the binding of [35S]GTPγS to G protein in membranes obtained from CHO cells stably-expressing human CB1R and treated with c) CP55,940 and 1 μM GAT compound, or d) GAT compound alone. Ki (nM) and EC50 (nM) are mean with 95% CI. EMax data are mean ± SEM. Data were fit to a variable slope (4 parameter) nonlinear regression model, mean ± SEM, n = 4–6 independent experiments performed in duplicate. ***P < 0.001 Student’s one sample t-test.

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques: Binding Assay, Stable Transfection, Expressing

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: (a-c) CHO cells stably-expressing hCB1R were treated with 0.10 nM – 10 μM GAT compounds alone for 90 min and cAMP inhibition (a) or βarrestin2 recruitment was measured (b). Compound agonist bias between cAMP inhibition and βarrestin2 recruitment (ΔΔLogR (cAMP - βarr2)] is shown in c. (d-f) CHO cells stably-expressing hCB1R were treated with 100 nM CP55,940 + 0.10 nM – 10 μM GAT compounds for 90 min and cAMP inhibition (d) or βarrestin2 recruitment was measured (e). Compound PAM bias between cAMP inhibition and βarrestin2 recruitment (ΔΔLogR (cAMP - βarr2)] is shown in f. cAMP data are expressed as % Forskolin response within compound treatment. βarrestin2 recruitment data are expressed as % CP55,940 response. Data were fit to a nonlinear regression (4 parameter model, GraphPad v. 7) for statistics in Table 5; or fit to the operational model (eq. 1) to calculate bias (c,f). Data are mean ± S.E.M. or 95% CI (c,f), n ≥ 6 independent experiments performed in triplicate.

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques: Stable Transfection, Expressing, Inhibition

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: The CB1 positive allosteric modulators 6r and 6s suppress CFA-Induced mechanical allodynia. (A, B) Both 6r (0.1, 0.3, 1, 3, 10 and 30 mg/kg i.p.) and 6s (0.1, 0.3, 1, 3, 10 and 30 mg/kg i.p.) increase mechanical paw withdrawal thresholds in the paw ipsilateral (A) but not contralateral (B) to CFA injection. (C, D) Both 6r (30 mg/kg i.p.) and 6s (30 mg/kg i.p.) elevated mechanical paw withdrawal thresholds in the paw ipsilateral (C) but not contralateral (D) to CFA injection with efficacy observed over at least 2.5 h post injection. (E, F) Neither 6r nor 6s altered mechanical paw withdrawal thresholds in CB1 KO mice in the paw ipsilateral (E) or contralateral (F) to CFA injection.

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques: Injection

Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: Effect of key CB1R ago-PAMs on catalepsy, hypothermia and tail flick test. Male C57BL/6 mice treated with 0.1 – 10 mg/kg compound or vehicle (1:1:18 ethanol:cremaphor:saline) and physiological assessments of catalepsy (5 min, % MPE 60 sec) (a), body temperature relative to vehicle (15 min, 100%) (b), and nociception in the tail flick assay (52°C) (20 min, % MPE 20 sec) (c). n = 5–6, data are mean ± SEM. *P < 0.05, **P < 0.01 6r compared to vehicle; ^P < 0.05, ^^^P < 0.001 6s compared to vehicle as determined via two-way ANOVA followed by Tukey’s post-hoc test.

Article Snippet: PathHunter® CB1R βarrestin2 assay: βarrestin2 recruitment was determined using the hCB1R CHO-K1 cell PathHunter® assay (DiscoveRx®).

Techniques: Tail Flick Test, Saline