Journal: Journal of Hypertension

Article Title: Vasodilatory effects of cannabidiol in human pulmonary and rat small mesenteric arteries: modification by hypertension and the potential pharmacological opportunities

doi: 10.1097/hjh.0000000000002333

Figure Lengend Snippet: FIGURE 3 Mechanisms of the cannabidiol-induced vasorelaxation in isolated human pulmonary arteries. Concentration–response curve of the cannabidiol-induced vaso- relaxation after 30 min incubation (before precontraction) with antagonists of CB1 AM251 (1 mmol/l), CB2 AM630 (1 mmol/l) (a) and endothelium-dependent relaxation to cannabinoids O-1918 (10 mmol/l), TRPV1 capsazepine (1 mmol/l) and TRPV4 RN1734 (20 mmol/l) (b) in endothelium-intact hPAs. The results are presented as the mean SEM of n tissue samples for each curve. In a few cases, SEM is smaller than or equal to the size of symbols. See Table 2 for n and the statistical analysis if not stated here. Representative CB1 (c), CB2 (d), TRPV1 (e) and GPR18 (f) immunoreactivity. Scale bars: 50 mm. hPAs, human pulmonary arteries; SEM, standard error of the mean.

Article Snippet: In the immunohistochemical study, the EnVision method was used according to Baranowska-Kuczko et al. [25] using an antibody against the cannabinoid CB1 receptor [1: 1000; rabbit Anti-CB1 (no cat. ab23703); Abcam, Cambridge, UK], cannabinoid CB2 receptor [1: 200; rabbit Anti-CB2 (no cat. ab3561); Abcam], TPRV1 [1: 1000; rabbit Anti-TRPV1 (no cat. bs-1931R); Bioss Antibodies, Woburn, Massachusetts, USA] and GPR18 [1: 1000; rabbit Anti-GPR18 (no cat. ab150618); Abcam].

Techniques: Isolation, Concentration Assay, Incubation

Journal: Journal of Hypertension

Article Title: Vasodilatory effects of cannabidiol in human pulmonary and rat small mesenteric arteries: modification by hypertension and the potential pharmacological opportunities

doi: 10.1097/hjh.0000000000002333

Figure Lengend Snippet: FIGURE 6 Mechanisms of the cannabidiol-induced vasorelaxation in isolated rat small mesenteric arteries. Concentration–response curve of the cannabidiol-induced vasorelaxation after 30 min incubation (before precontraction) with antagonists of CB1 AM251 (1 mmol/l; a–h), CB2 AM630 (1 mmol/l; a–d), TRPV1 capsazepine (1 mmol/l; a–d) in endothelium-intact (a–d) and endothelium-denuded (-ENDO; e–h) rat sMAs. The results are presented as the mean SEM of n tissue samples for each curve. See Table 3 for n and the statistical analysis. In a few cases, SEM is smaller than or equal to the size of symbols. SEM, standard error of the mean; sMAs, small mesenteric arteries.

Article Snippet: In the immunohistochemical study, the EnVision method was used according to Baranowska-Kuczko et al. [25] using an antibody against the cannabinoid CB1 receptor [1: 1000; rabbit Anti-CB1 (no cat. ab23703); Abcam, Cambridge, UK], cannabinoid CB2 receptor [1: 200; rabbit Anti-CB2 (no cat. ab3561); Abcam], TPRV1 [1: 1000; rabbit Anti-TRPV1 (no cat. bs-1931R); Bioss Antibodies, Woburn, Massachusetts, USA] and GPR18 [1: 1000; rabbit Anti-GPR18 (no cat. ab150618); Abcam].

Techniques: Isolation, Concentration Assay, Incubation

Journal: Journal of Hypertension

Article Title: Vasodilatory effects of cannabidiol in human pulmonary and rat small mesenteric arteries: modification by hypertension and the potential pharmacological opportunities

doi: 10.1097/hjh.0000000000002333

Figure Lengend Snippet: FIGURE 7 The relative Cnr1 (a) and Cnr2 (b) gene expression evaluated by real-time quantitative PCR in small mesenteric arteries of hypertensive SHR and DOCA-salt vs. normotensive controls WKY and SHAM, respectively. Results are shown mean SEM (n ¼ 6 per group) as relative fold change in mRNA expression in comparison to respective control, where expression level was set as 1. P < 0.01 or P < 0.001 according to the Student’s t-test. DOCA-salt, 11-deoxycorticosterone acetate-salt; SEM, standard error of the mean; SHR, spontaneously hypertensive rats; SHAM, control sham-operated rats; WKY, Wistar–Kyoto.

Article Snippet: In the immunohistochemical study, the EnVision method was used according to Baranowska-Kuczko et al. [25] using an antibody against the cannabinoid CB1 receptor [1: 1000; rabbit Anti-CB1 (no cat. ab23703); Abcam, Cambridge, UK], cannabinoid CB2 receptor [1: 200; rabbit Anti-CB2 (no cat. ab3561); Abcam], TPRV1 [1: 1000; rabbit Anti-TRPV1 (no cat. bs-1931R); Bioss Antibodies, Woburn, Massachusetts, USA] and GPR18 [1: 1000; rabbit Anti-GPR18 (no cat. ab150618); Abcam].

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Expressing, Comparison, Control

Journal: Journal of Hypertension

Article Title: Vasodilatory effects of cannabidiol in human pulmonary and rat small mesenteric arteries: modification by hypertension and the potential pharmacological opportunities

doi: 10.1097/hjh.0000000000002333

Figure Lengend Snippet: FIGURE 8 Representative micrographs of immunohistochemical staining of CB1, CB2, TRPV1 and GPR18 receptors in cross-sections of rat small mesenteric arteries from normotensive control (WKY; SHAM) and hypertensive (SHR; DOCA-salt) rats. Scale bars: 50 mm. DOCA-salt, 11-deoxycorticosterone acetate-salt; SHR, spontaneously hypertensive rats; SHAM, control sham-operated rats; WKY, Wistar–Kyoto.

Article Snippet: In the immunohistochemical study, the EnVision method was used according to Baranowska-Kuczko et al. [25] using an antibody against the cannabinoid CB1 receptor [1: 1000; rabbit Anti-CB1 (no cat. ab23703); Abcam, Cambridge, UK], cannabinoid CB2 receptor [1: 200; rabbit Anti-CB2 (no cat. ab3561); Abcam], TPRV1 [1: 1000; rabbit Anti-TRPV1 (no cat. bs-1931R); Bioss Antibodies, Woburn, Massachusetts, USA] and GPR18 [1: 1000; rabbit Anti-GPR18 (no cat. ab150618); Abcam].

Techniques: Immunohistochemical staining, Staining, Control

Journal: Cerebral Cortex (New York, NY)

Article Title: Differential effects of group III metabotropic glutamate receptors on spontaneous inhibitory synaptic currents in spine-innervating double bouquet and parvalbumin-expressing dendrite-targeting GABAergic interneurons in human neocortex

doi: 10.1093/cercor/bhac195

Figure Lengend Snippet: Primary antibody information.

Article Snippet: CB1 , rabbit , Immunogenes , Anti-CB1 (Rabbit) Polyclonal Antibody , ( ) , , .

Techniques: Protein Concentration, Recombinant, Bacteria, Binding Assay, Purification

Journal: Cerebral Cortex (New York, NY)

Article Title: Neural stem cell lineage-specific cannabinoid type-1 receptor regulates neurogenesis and plasticity in the adult mouse hippocampus

doi: 10.1093/cercor/bhy258

Figure Lengend Snippet: (A) Scheme of the strategy for conditional deletion of CB1 from NSCs and genetic tagging of NSC with YFP in vivo (using nes-CreERT2, expressing tamoxifen-inducible Cre under the control of nestin promoter/enhancer elements, P/E), obtaining nes-CB1ko/ko. The respective controls (CB1wt/wt) contain the wild-type CB1 alleles, but are YFP tagged. (B) Time course of experiment. Mice were perfused at 28 days post tamoxifen-induced recombination (dptm) or 56 dptm. (C) Recombined cells express YFP (green) and are present in the subventricular zone (SVZ) and (D) in the subgranular zone (SGZ) of the dentate gyrus (DG). (E) Quantification of YFP-positive cells in the DG revealed a significant decrease in nes-CB1ko/ko mice as compared with CB1wt/wt at 28 dptm and 56 dptm. n = 4 animals/group, ** P < 0.01, * P < 0.05, two-tailed unpaired Student’s t-test. Data are represented as mean ± SEM. (F) Representative confocal images including z-stacks display co-localization of recombined YFP cells (green) and CB1 expression (red) in CB1wt/wt mice, whereas nes-CB1ko/ko mice show a lack of CB1 expression in recombined YFP cells (arrowheads indicate corresponding points in the orthogonal planes). DAPI, blue. Scale bar, 100 μm. Cortex (Cx); Striatum (Str); granule cell layer (GCL), molecular layer (ML).

Article Snippet: The following primary antibodies were used: rat anti-BrdU (1:100, Abcam), rabbit anti-GFP (1:500, kind gift of Matthias Klugman, Sydney, Australia), chicken anti-GFP (1:1000, Aves Labs), rabbit anti-RFP (1:500, Rockland), guinea pig anti-DCX (1:500, Merck), mouse anti-NeuN (1:500, Abcam), rabbit anti-S100b (1:500, Abcam), rabbit anti-CB1 (1:1000, Immunogenes), rabbit anti-CALB (1:5000, Swant), rabbit anti-cleaved Casp3 (1:200, Cell Signaling), mouse anti-GAD67 (1:500, Millipore).

Techniques: In Vivo, Expressing, Two Tailed Test

Journal: Cerebral Cortex (New York, NY)

Article Title: Neural stem cell lineage-specific cannabinoid type-1 receptor regulates neurogenesis and plasticity in the adult mouse hippocampus

doi: 10.1093/cercor/bhy258

Figure Lengend Snippet: (A) Schedule of experiment. Mice were injected with BrdU for five d at 23 dptm and were analyzed at 28 dptm and at 56 dptm, respectively. (B) Maximum intensity projections of z-stacks show YFP-positive cells expressing DCX, NeuN or S100b at 28 dptm. (C) Maximum intensity projections of z-stacks show YFP expressing cells positive for BrdU at 28 dptm (filled arrowhead: colocalizing cell). (D) Triple co-localization (filled arrowhead) at 56 dptm (with magnification in the quadrant), empty arrowhead: positive YFP/BrdU cell. (E) Quantification of YFP-positive cells co-localizating with the neuronal and astrocytic markers. Deletion of CB1 reduces the YFP/DCX and YFP/NeuN population at 28 dptm and 56 dptm. YFP/S100b cells were significantly reduced only at 28 dptm. n = 5 (28 dptm S100b), n = 3 (28 dptm DCX), n = 4 (28 dptm NeuN), n = 4 (56 dptm S100b), n = 4 (56 dptm DCX), n = 4 (56 dptm NeuN) animals/group * P < 0.05, ** P < 0.01, two-tailed unpaired Student’s t-test. (F) Nes-CB1ko/ko animals displayed a significantly reduced number of newborn cells, as assessed by co-localization of YFP with BrdU. n = 5 (28 dptm CB1wt/wt), n = 6 (28 dptm nes-CB1ko/ko), n = 4 (56 dptm CB1wt/wt), n = 4 (28 dptm nes-CB1ko/ko) animals/group; * P < 0.05, ** P < 0.01, two-tailed unpaired Student’s t-test. (G) Differentiation of BrdU labeled cells into the neuronal (NeuN) or astroglial (S100b) lineage is not affected at 56 dptm. N = 4 animals/group, two-tailed unpaired Student’s t-test, n.s. not significant. Data represent mean ± SEM. Scale bar, 50 μm. Subgranular zone (SGZ), granule cell layer (GCL), molecular layer (ML).

Article Snippet: The following primary antibodies were used: rat anti-BrdU (1:100, Abcam), rabbit anti-GFP (1:500, kind gift of Matthias Klugman, Sydney, Australia), chicken anti-GFP (1:1000, Aves Labs), rabbit anti-RFP (1:500, Rockland), guinea pig anti-DCX (1:500, Merck), mouse anti-NeuN (1:500, Abcam), rabbit anti-S100b (1:500, Abcam), rabbit anti-CB1 (1:1000, Immunogenes), rabbit anti-CALB (1:5000, Swant), rabbit anti-cleaved Casp3 (1:200, Cell Signaling), mouse anti-GAD67 (1:500, Millipore).

Techniques: Injection, Expressing, Two Tailed Test, Labeling

Journal: Cerebral Cortex (New York, NY)

Article Title: Neural stem cell lineage-specific cannabinoid type-1 receptor regulates neurogenesis and plasticity in the adult mouse hippocampus

doi: 10.1093/cercor/bhy258

Figure Lengend Snippet: (A) Maximum intensity projections of z-stacks showing YFP- positive cells expressing calretinin (CALR) at 28 dptm (arrowhead: co-localizing cell) and calbindin (CALB) at 56 dptm. Scale bar, 20 μm. (B) Quantification of YFP cells co-localizing with the two stage-specific markers. Deletion of CB1 reduced the YFP/CALR-positive population at 28 dptm and YFP/CALB-positive population at 56 dptm. N = 4 animals/group * P < 0.05, ** P < 0.01, two-tailed unpaired Student’s t-test. (C) Maximum intensity projections of z-stacks showing YFP/BrdU-positive cells in the SVZ at 28 dptm, overview and magnified area of the rectangle in the overview. Scale bar, 50 μm. (D) Quantification of anti-BrdU and anti-YFP co-staining in the SVZ demonstrates no difference between CB1wt/wt and nes-CB1 ko/ko animals. ( n = 4 for CB1wt/wt, n = 5 for nes-CB1ko/ko). (E) Exemplary micrographs showing apoptotic cells evaluated by activated caspase-3 (Casp3) staining and YFP-positive cells in CB1wt/wt and nes-CB1ko/ko mice at 28 dptm. Scale bar, 20 μm. (F) Averaged number of Casp3-positive cells in the DG per hippocampal section (left panel) and percentage of YFP/Casp3-positive cells of all Casp3-positive cells per section (right panel) at 28 dptm. N = 4 animals per genotype (CB1wt/wt: n = 13 sections; nes-CB1ko/ko: n = 19 sections).

Article Snippet: The following primary antibodies were used: rat anti-BrdU (1:100, Abcam), rabbit anti-GFP (1:500, kind gift of Matthias Klugman, Sydney, Australia), chicken anti-GFP (1:1000, Aves Labs), rabbit anti-RFP (1:500, Rockland), guinea pig anti-DCX (1:500, Merck), mouse anti-NeuN (1:500, Abcam), rabbit anti-S100b (1:500, Abcam), rabbit anti-CB1 (1:1000, Immunogenes), rabbit anti-CALB (1:5000, Swant), rabbit anti-cleaved Casp3 (1:200, Cell Signaling), mouse anti-GAD67 (1:500, Millipore).

Techniques: Expressing, Two Tailed Test, Staining

Journal: Cerebral Cortex (New York, NY)

Article Title: Neural stem cell lineage-specific cannabinoid type-1 receptor regulates neurogenesis and plasticity in the adult mouse hippocampus

doi: 10.1093/cercor/bhy258

Figure Lengend Snippet: (A) Scheme of deletion of CB1 from neural stem cells in vivo with a lentivirus containing Cre recombinase under the control of the Sox2 promoter (Sox2 P), obtaining Sox2-CB1ko/ko. The respective controls (CB1wt/wt) contain the wild-type CB1 alleles, but are YFP tagged. (B) Experimental timeline. Eight weeks old animals were injected with the lentivirus (LV) via stereotactic surgery. BrdU was injected i.p. for five consecutive days starting at 23 dpi, and animals were perfused at 28 dpi and 56 dpi. (C) Representative micrographs of CB1wt/wt and Sox2-CB1ko/ko mice show recombined cells in the subgranular zone (SGZ) expressing YFP (green) after lentivirus injection and a fraction of it co-localizing with BrdU (red) at 28 dpi and at 56 dpi. (D) Quantification of YFP/BrdU double positive cells showed that CB1 loss impaired significantly cell proliferation (at 28 dpi), but maintenance of YFP/BrdU cells (at 56 dpi) is only slightly (but not significantly) decreased upon CB1 deletion ( P = 0.060). N = 3 animals/group, * P < 0.05, two-tailed unpaired Student’s t-test, t = 2.845. (E) Exemplary micrographs of CB1wt/wt animals for YFP (green) cells co-localizing with BrdU (red) and S100b or NeuN (blue) in the dentate gyrus at 56 dpi. Arrowheads mark triple positive cells. (F) Percentage of YFP/BrdU/marker of total YFP/BrdU-positive cells (marker: S100b or NeuN) at 56 dpi. Differentiation into the astroglial or neuronal lineage is not affected. N = 4 (CB1wt/wt) and n = 3 (Sox2-CB1ko/ko) animals/group, n.s. not significant in two-tailed unpaired Student’s t-test. Values represent mean ± SEM. Scale bar, 100 μm.

Article Snippet: The following primary antibodies were used: rat anti-BrdU (1:100, Abcam), rabbit anti-GFP (1:500, kind gift of Matthias Klugman, Sydney, Australia), chicken anti-GFP (1:1000, Aves Labs), rabbit anti-RFP (1:500, Rockland), guinea pig anti-DCX (1:500, Merck), mouse anti-NeuN (1:500, Abcam), rabbit anti-S100b (1:500, Abcam), rabbit anti-CB1 (1:1000, Immunogenes), rabbit anti-CALB (1:5000, Swant), rabbit anti-cleaved Casp3 (1:200, Cell Signaling), mouse anti-GAD67 (1:500, Millipore).

Techniques: In Vivo, Injection, Expressing, Two Tailed Test, Marker

Journal: World Journal of Gastroenterology

Article Title: Anti-inflammatory effect of cannabinoid agonist WIN55, 212 on mouse experimental colitis is related to inhibition of p38MAPK

doi: 10.3748/wjg.v22.i43.9515

Figure Lengend Snippet: Representative figures of the expression of cannabinoid receptor type 1 and type 2 receptors in mouse colon by immunohistochemistry staining, which are from 4 separate experiments with similar results. A: Expression of cannabinoid (CB)1 receptors; B: Expression of CB2 receptors; (magnification × 400, respectively). The expressions of CB1 receptor in the colonic epithelial cells and CB2 receptor in the submucosal immune cells are marked by arrows and presented as positive results with brown-yellow staining.

Article Snippet: The first antibody was rabbit anti-CB1 receptor or rabbit anti-CB2 receptor polyclonal antibody (1:20 dilution for both; Enzo, Plymouth Meeting, PA, United States).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: World Journal of Gastroenterology

Article Title: Anti-inflammatory effect of cannabinoid agonist WIN55, 212 on mouse experimental colitis is related to inhibition of p38MAPK

doi: 10.3748/wjg.v22.i43.9515

Figure Lengend Snippet: Expression of cannabinoid receptor type 1 by western blot analysis. A: Collection of colonic tissues specimens from C57BL/6 mice in different experimental groups and the western blotting operation were performed as described in the Materials and Methods; B: Representative strips from 4 separate experiments with similar results are presented. Semi-quantitative blot analysis of cannabinoid (CB)1 receptors was carried out by ImageJ software. Data are presented as mean ± SD ( n = 4). a P < 0.05 vs control; c P < 0.05 vs dextran sulfate sodium (DSS) + vehicle (Veh).

Article Snippet: The first antibody was rabbit anti-CB1 receptor or rabbit anti-CB2 receptor polyclonal antibody (1:20 dilution for both; Enzo, Plymouth Meeting, PA, United States).

Techniques: Expressing, Western Blot, Software