Journal: Journal of Inflammation (London, England)

Article Title: Cannabinoid 2 receptor attenuates inflammation during skin wound healing by inhibiting M1 macrophages rather than activating M2 macrophages

doi: 10.1186/s12950-018-0201-z

Figure Lengend Snippet: a and b Representative immunoblotting results and ( c ) semi-quantitative analysis of CB2R protein level at different posttraumatic intervals. + P < 0.05, ++ P < 0.01, JWH133 versus vehicle; * P < 0.05, ** P < 0.01, GP1a versus vehicle; # P < 0.05, ## P < 0.01, AM630 versus vehicle

Article Snippet: After being blocked with 5% fat-free milk at RT for 2 h, PVDF membranes were incubated, respectively, with a rabbit anti-CB2R polyclonal primary antibody (1:250 dilution; sc-25,494, Santa Cruz Biotechnology, CA, USA) and mouse anti–GAPDH monoclonal antibody (1:2500 dilution, sc-32,233; Santa Cruz Biotechnology, Inc) at 4 °C overnight.

Techniques: Western Blot

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: HIV-infected MDM keep expressing surface CB2R at day 12pi. MDM were labeled with a fluorescent CB2R monoclonal antibody (FL4 channel) conjugated with Alexa Fluor® 647 and then analyzed by flow cytometry. ( a ) Representative histograms of extracellular staining show significant differences in fluorescence intensities from HIV + (dark histogram) and uninfected (grey histogram) on the expression of CB2R compared with the nonfluorescent MDM (unshaded histograms to the left. ( b ) A graphic representation of surface CB2R levels per donor is shown. Panels a and b are representative of three different donors (n = 3). An unstained control was included as a technical negative control and is representative of 1 donor (n = 1). ( c ) Representative images of intracellular levels of CB2R and GAPDH expression using Western Blot. Images were cropped from the same blot after stripping and reprobing with a different antibody and were acquired using different exposure times. Full-length blots are presented in Supplementary Fig. 4 ( d ) A graphic representation of intracellular CB2R levels per donor is shown. Figures c and d are representative of four different donors (n = 4).

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Infection, Expressing, Labeling, Flow Cytometry, Staining, Fluorescence, Control, Negative Control, Western Blot, Stripping Membranes

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: CB2R agonists treatment decrease HIV-1 replication and CATB secretion from MDM. MDM were infected with HIV-1 and treated with CB2R agonists, JWH-133 and HU-308. HIV-1 p24 and CATB levels were measured from supernatants of days 3, 6, 9, and 12pi by ELISA. ( a ) Temporal HIV-1 p24 levels in supernatants of HIV-infected MDM treated with JWH-133. ( b ) Temporal CATB secretion levels in supernatants of HIV-infected MDM treated with JWH-133. Figures a and b are representative of at least four different donors (n = at least 4). ( c ) Temporal HIV-1 p24 levels in supernatants of HIV-infected MDM treated with HU-308. ( d ) Temporal CATB secretion levels in supernatants of HIV-infected MDM treated with HU-308. Figures c and d are representative of at least six different donors (n = at least 6). Graphs are presented using the mean and the ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01 vs. vehicle control at its respective time-point.

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: JWH-133 prevents HIV-induced increase in surface CB2R expression and induces oscillating expressions over time. MDM were cultured in 8-well chamber slides, infected with HIV-1 ADA , and treated with JWH-133 at 0.5 µM. Slides were fixed at days 3, 6, 9, and 12dpi. Cells were stained with an anti-CB2R antibody (red) and nuclei were stained with DAPI (blue). At least 3 different random pictures per condition were acquired. Representative immunofluorescence images are shown. Graphs are presented using the mean ± standard error of the mean (SEM). This figure is representative of three different donors (n = 3). * p < 0.05, ** p < 0.01.

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Expressing, Cell Culture, Infection, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: JWH-133 decreases HIV-1 replication and CATB secretion through CB2R activation. MDM were cultured in 24-wells plates and infected with HIV-1 ADA . After removal of residual virus, cells were treated with CB2R antagonist SR144528 (SR: 1 µM) for 1 h, followed by JWH-133 (JWH) treatment at 0.5 µM. Treatments were maintained for 6dpi, exchanging half of the media at day 3pi and repeating the co-administration protocol. HIV-1 p24 and CATB levels were measured in HIV-infected MDM supernatants using ELISA. HIV-1 p24 and CATB levels were normalized against its vehicle control per donor. ( a ) HIV-1 p24 levels in HIV-infected MDM after co-administration of CB2R ligands. This figure is representative of at least eight different donors (n = 8). ( b ) CATB levels in HIV-infected MDM after co-administration of CB2R ligands. This figure is representative of ten different donors (n = 10). Graphs are presented using the mean ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01.

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Activation Assay, Cell Culture, Infection, Virus, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of neurochemistry

Article Title: Δ9-THC increases endogenous AHA1 expression in rat cerebellum and may modulate CB1 receptor function during chronic use

doi: 10.1111/j.1471-4159.2011.07391.x

Figure Lengend Snippet: Proteins differentially regulated by chronic Δ9-THC in intact and ovarectomized rats DeCyder software (GE Healthcare) was used for simultaneous comparison of abundance changes across all samples’ 2-DE gels, and for comparisons of individual Cy3 and Cy5 samples for each rat. The DeCyder differential in-gel analysis (DIA) module was used for pair-wise comparisons of each sample on a gel to the Cy2 labeled standard present on each gel. For each pair-wise DIA comparison, the entire signals from each CyDye channel are normalized prior to the co-detection of protein spot boundaries and the calculation of the volume ratio for each protein spot. The DeCyder biological variation analysis (BVA) module was then used to simultaneously match all protein spot maps from all gels, and using the Cy3/Cy5:Cy2 DIA ratios, calculate abundance changes and paired Student’s t -test p -values for the variance of these ratios for each protein pair across all samples. In the absence of sufficient experimental replicates, no statistical information can be generated. Fold abundance changes are reported, whereby a fold increase is calculated directly from the volume ratio, and a fold decrease by the inverse of volume ratio. The DIA module was also used to calculate the direct Cy5:Cy3 volume ratio for each paired sample individually (without the Cy2 mixed standard) to assess the contribution from each subject and reveal changes that were present in a group. After determining spots of interest, the protein were excised using the Ettan Spot Handling Workstation with a 2-mm diameter spot-picking head (GE Healthcare). Gel spots were cut, de-stained and then digested with trypsin (Promega). The resulting peptide mixture was loaded on a Dionex PepMap C18 trap column and was separated by a New Objective reversed phase C18 Picofrit column/emitter. Peptide mass was determined by a Thermo-Fisher LTQXL linear ion trap mass spectrometer (Waltham, MA, USA) coupled with an Eksigent nanoLC (Dublin, CA, USA). The raw data were analyzed by the Mascot search engine V2.2 (Matrix Science Inc, Boston, MA, USA) against the rat SwissProt database (false discovery rate <5%) to generate a list of possible proteins for that gel spot.

Article Snippet: The sources of the antibodies used in the present study were as follows: anti-CB1R and anti-CB2R antibodies were from Cayman, anti-hemagglutinin (anti-HA) was from Santa Cruz, anti-AHA1 antibody was from Abnova, anti-HSP90α and anti-HSP90β antibodies were from Enzo Lifesciences, secondary anti-mouse and anti-rabbit antibodies were from Perkin Elmer, Alexa Fluor 594-labeled anti-mouse and anti-rabbit and anti-mouse antibodies and 4,6-diamidino-2-phenylindole were obtained from Invitrogen.

Techniques: Software, Comparison, Labeling, Generated, Mass Spectrometry, Clinical Proteomics, Membrane

Journal: Journal of neurochemistry

Article Title: Δ9-THC increases endogenous AHA1 expression in rat cerebellum and may modulate CB1 receptor function during chronic use

doi: 10.1111/j.1471-4159.2011.07391.x

Figure Lengend Snippet: A. Western blots of AHA1 levels in the cerebellar extracts from intact or ovariectimized female rats administered either saline or Δ9-THC during adolescence. The cerebellar extracts (20 μg/lane) were separated by 10% SDS-PAGE, transferred onto PDVF membranes, and subjected to western-blotting with specific antibodies. The data were quantified using a Fuji Film luminescent image analyzer (LAS-1000 Plus) and the Image Gauge program (Version 3.4). B. Quantification for the experiments shown in A. AHA1 levels were corrected for the β-actin levels in the same samples and the data are presented as the % from the levels found in the control group. *- indicate statistical significant differences between the four groups with p < 0.05.

Article Snippet: The sources of the antibodies used in the present study were as follows: anti-CB1R and anti-CB2R antibodies were from Cayman, anti-hemagglutinin (anti-HA) was from Santa Cruz, anti-AHA1 antibody was from Abnova, anti-HSP90α and anti-HSP90β antibodies were from Enzo Lifesciences, secondary anti-mouse and anti-rabbit antibodies were from Perkin Elmer, Alexa Fluor 594-labeled anti-mouse and anti-rabbit and anti-mouse antibodies and 4,6-diamidino-2-phenylindole were obtained from Invitrogen.

Techniques: Western Blot, Saline, SDS Page, Control

Journal: Journal of neurochemistry

Article Title: Δ9-THC increases endogenous AHA1 expression in rat cerebellum and may modulate CB1 receptor function during chronic use

doi: 10.1111/j.1471-4159.2011.07391.x

Figure Lengend Snippet: A. AHA-1 levels in control and AHA-1 transfected HEK293T cells. The cells were transfected with pcDNA 3.1 or AHA-1 (2.25 μg/well in 6-well plates) and serum starved for 24 h. Two days after transfection the cells were lysated and 10 μg of total protein were separated by 10% SDS-PAGE and AHA-1 levels were determined by western blot. B. Modulation of CB1R and CB2R plasma membrane levels by AHA1 overexpression. HEK293T were co-transfected with CB1R or CB2R (0.25 μg/well) and pcDNA 3.1 or AHA-1 (2.25 μg/well each) and after six hours the cells were trypsinized and plated on 12-well plates as described in Material and Methods. FBS was withdrawn for 24 h, and two days after transfection the plasma membrane levels of CB1R and CB2R were determined by ELISA as described in Material and Methods. n=12 in each case from three different transfections. *- indicate p < 0.05 compared to pcDNA 3.1 transfected cells. C. Subcellular localization of CB1R in HEK293T cells. The cells were grown on coverslips in 6-well plates and transfected with 3xHA-CB1R (0.1 μg/well) and DsRed-Rab5 (0.1 μg/well). After serum starvation for 24 h, the cells were fixed and permeabilized. The CB1R localization was stained using HA antibody as described in Material and Methods. Blue: DNA staining by 4,6-diamidino-2-phenylindole (nuclear), green: 3xHA-CB1R, red: DsRed-Rab5. The images are representative from three different coverslips, obtained from three independent transfections. D. CB1R interacts with AHA-1 in HEK293T cells. The cells were transiently co-transfected in 10 mm2 dishes with pcDNA 3.1 (10 μg, control), or with CB1R (5 μg) and pcDNA3.1 (5 μg), or CB1R (5 μg) and AHA-1 (5 μg). After two days the cells were solubilized and immunoprecipitated with CB1R antibody as described under Material and Methods. The immunoprecipitates (20 μg/lane) or the lysates (10 μg/lane) were separated by 10 % SDS-Page and subject to western-blotting with AHA-1 and CB1R antibodies. The experiment shown is representative from three independent transfections.

Article Snippet: The sources of the antibodies used in the present study were as follows: anti-CB1R and anti-CB2R antibodies were from Cayman, anti-hemagglutinin (anti-HA) was from Santa Cruz, anti-AHA1 antibody was from Abnova, anti-HSP90α and anti-HSP90β antibodies were from Enzo Lifesciences, secondary anti-mouse and anti-rabbit antibodies were from Perkin Elmer, Alexa Fluor 594-labeled anti-mouse and anti-rabbit and anti-mouse antibodies and 4,6-diamidino-2-phenylindole were obtained from Invitrogen.

Techniques: Control, Transfection, SDS Page, Western Blot, Clinical Proteomics, Membrane, Over Expression, Enzyme-linked Immunosorbent Assay, Staining, Immunoprecipitation

Journal: Journal of neurochemistry

Article Title: Δ9-THC increases endogenous AHA1 expression in rat cerebellum and may modulate CB1 receptor function during chronic use

doi: 10.1111/j.1471-4159.2011.07391.x

Figure Lengend Snippet: HEK293T co-transfected with CB1R or CB2R and pcDNA3.1 or AHA1 as in Fig 2B were plated on 24 well-plates and serum straved for 24h. Two days after transfection the medium was changed to PBS containing 100 μM IBMX for one hour. Subsequently, the cells were pretreated with Δ9-THC (1 μM)) for 5 min, followed by stimulation with forskolin (10 μM) for 15 min. The reactions were stopped by medium aspiration and addition of 200 μl thhricloracetic acid. cAMP levels were determined using cAMP Elisa Kit (Cayman Biochemicals) as described in Material and Methods. n=12 in each case from three independent transfections. *- indicates statistically significant differences between pcDNA 3.1 and AHA1 transfected cells by two-way ANOVA followed by Holm Sidak-test (interaction: F(2,12=14.03, p<0.001).

Article Snippet: The sources of the antibodies used in the present study were as follows: anti-CB1R and anti-CB2R antibodies were from Cayman, anti-hemagglutinin (anti-HA) was from Santa Cruz, anti-AHA1 antibody was from Abnova, anti-HSP90α and anti-HSP90β antibodies were from Enzo Lifesciences, secondary anti-mouse and anti-rabbit antibodies were from Perkin Elmer, Alexa Fluor 594-labeled anti-mouse and anti-rabbit and anti-mouse antibodies and 4,6-diamidino-2-phenylindole were obtained from Invitrogen.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay

Journal: Journal of neurochemistry

Article Title: Δ9-THC increases endogenous AHA1 expression in rat cerebellum and may modulate CB1 receptor function during chronic use

doi: 10.1111/j.1471-4159.2011.07391.x

Figure Lengend Snippet: HEK293T cells in 6-well plates were co-transfected with CB1R (0.25/μg/well) and pcDNA 3.1 or AHA1 (each at 2.25 μg/well). Subsequently, the cells were serum starved for 24 h and then stimulated with increasing concentrations of Δ9-THC for 5 min. The reactions were stopped by aspiration of the medium and addition of 200 μl lysis buffer. Ten μg of protein were separated by SDS-page and ERK 1/2 activation and total ERK 1/2 levels were detected using specific antibodies. Top panel: representative blots from four different experiments. Lower panel: Quantification of the effects of Δ9-THC on MAPK levels, open symbols indicate pcDNA 3.1 transfected cells and black symbols indicate AHA1 overexpressing cells; n=4 in each case from four different transfections, * indicates statistically significant differences compared to pcDNA 3.1 transfected cells by one-way ANOVA.

Article Snippet: The sources of the antibodies used in the present study were as follows: anti-CB1R and anti-CB2R antibodies were from Cayman, anti-hemagglutinin (anti-HA) was from Santa Cruz, anti-AHA1 antibody was from Abnova, anti-HSP90α and anti-HSP90β antibodies were from Enzo Lifesciences, secondary anti-mouse and anti-rabbit antibodies were from Perkin Elmer, Alexa Fluor 594-labeled anti-mouse and anti-rabbit and anti-mouse antibodies and 4,6-diamidino-2-phenylindole were obtained from Invitrogen.

Techniques: Transfection, Lysis, SDS Page, Activation Assay

Journal: Journal of neurochemistry

Article Title: Δ9-THC increases endogenous AHA1 expression in rat cerebellum and may modulate CB1 receptor function during chronic use

doi: 10.1111/j.1471-4159.2011.07391.x

Figure Lengend Snippet: A. AHA-1 levels in control and AHA-1 transfected Neuro-2A cells. The cells were transfected with pcDNA 3.1 or AHA-1 (2.25 μg/well in 6-well plates) and serum starved for 24 h. The lysates and the western blot were performed as described in Fig 2A. B. Cell surface levels of CB1R in Neuro-2A overexpressing cells. Neuro2-A cells were transfected as above and then trypsinized and plated at a density of 50×104 cells/well in 12-well plates. Subsequent procedures to determine plasma membrane CB1R levels were similar to that described for HEK293T cells in Fig 2B. n=9 from three different experiments. * indicates statistically significant differences compared to pcDNA 3.1 transfected cells. C. The effects of CB1R stimulation on cAMP levels in pcDNA 3.1 or AHA-1 transfected Neuro-2A cells. The transfections were made as described and subsequently, the cAMP determinations were conducted as described in the Fig 3. n=12 in each case from three different transfections; * - indicates statistically significant differences between pcDNA 3.1 and AHA1 transfected cells by two-way ANOVA followed by Holm Sidak-test (interaction: F(2,12=30.98, p<0.001).

Article Snippet: The sources of the antibodies used in the present study were as follows: anti-CB1R and anti-CB2R antibodies were from Cayman, anti-hemagglutinin (anti-HA) was from Santa Cruz, anti-AHA1 antibody was from Abnova, anti-HSP90α and anti-HSP90β antibodies were from Enzo Lifesciences, secondary anti-mouse and anti-rabbit antibodies were from Perkin Elmer, Alexa Fluor 594-labeled anti-mouse and anti-rabbit and anti-mouse antibodies and 4,6-diamidino-2-phenylindole were obtained from Invitrogen.

Techniques: Control, Transfection, Western Blot, Clinical Proteomics, Membrane

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Effect of maternal high-fat diet on key components of the placental and hepatic endocannabinoid system

doi: 10.1152/ajpendo.00119.2017

Figure Lengend Snippet: Endocannabinoid receptor 2 (CB2R) protein expression in the maternal and fetal livers of high-fat diet (HFD; n = 11) and control (CTR; n = 10) animals. A–D: representative images of fetal livers from male fetuses (MF) in the CTR (n = 3) and HFD (n = 6) diet groups. E–H: the fetal livers from female fetuses (FF) in the CTR (n = 4) and HFD (n = 4) groups. I–L: maternal livers of dams with MF in the CTR (n = 3) and HFD (n = 5) groups. M–P: maternal liver sections from dams carrying FF in the CTR (n = 5) and HFD (n = 4) groups. Q–T: quantitative analyses. Blue arrows point to bile ducts, green arrows point to arteries, pink arrows point to central veins (CV), and yellow asterisks indicate the portal vein. Negative controls (omission of primary antibodies) are shown in A–P, insets (top right). Scale bars, 20 μm. The data are presented as means ± SE.

Article Snippet: Immunohistochemistry (IHC) was performed using a CB1R monoclonal primary antibody (cat. no. Img-CB1R-mab001; Immunogenes, Budakeszi, Hungary), a CB2R monoclonal primary antibody (cat. no. H00001269-M01; Novus Biologicals, Littleton, CO), and an anti-mouse secondary antibody (Vectastain ABC Kit anti-mouse IgG, cat. no. PK-4002; Vector Laboratories, Burlingame, CA).

Techniques: Expressing

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Effect of maternal high-fat diet on key components of the placental and hepatic endocannabinoid system

doi: 10.1152/ajpendo.00119.2017

Figure Lengend Snippet: Fetal and maternal hepatic expression of main endocannabinoid receptors and metabolizing enzymes in male and female fetuses near term of HFD and CTR mothers

Article Snippet: Immunohistochemistry (IHC) was performed using a CB1R monoclonal primary antibody (cat. no. Img-CB1R-mab001; Immunogenes, Budakeszi, Hungary), a CB2R monoclonal primary antibody (cat. no. H00001269-M01; Novus Biologicals, Littleton, CO), and an anti-mouse secondary antibody (Vectastain ABC Kit anti-mouse IgG, cat. no. PK-4002; Vector Laboratories, Burlingame, CA).

Techniques: Expressing

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Effect of maternal high-fat diet on key components of the placental and hepatic endocannabinoid system

doi: 10.1152/ajpendo.00119.2017

Figure Lengend Snippet: Protein expression of CB1R, CB2R, fatty acid amide hydrolase-1 (FAAH), diacylglyceride lipase A (DAGLα), monoacylglycerol lipase (MAGL), and cyclooxygenase-2 (COX-2) in the liver and placenta. A: image of Western blot of CB1R, CB2R, FAAH, MAGL, and COX-2 in the maternal liver (n = 3–6). B: image of Western blot of CB1R, CB2R, FAAH, DAGLα, MAGL, and COX-2 in the fetal liver (n = 4–6). C: image of Western blot analysis (CB1R, CB2R, FAAH, DAGL, MAGL, and COX-2) in the placenta (n = 4–6). CTR, control; HFD, high-fat diet; MF, male fetus; FF, female fetus; M, male; F, female.

Article Snippet: Immunohistochemistry (IHC) was performed using a CB1R monoclonal primary antibody (cat. no. Img-CB1R-mab001; Immunogenes, Budakeszi, Hungary), a CB2R monoclonal primary antibody (cat. no. H00001269-M01; Novus Biologicals, Littleton, CO), and an anti-mouse secondary antibody (Vectastain ABC Kit anti-mouse IgG, cat. no. PK-4002; Vector Laboratories, Burlingame, CA).

Techniques: Expressing, Western Blot

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Effect of maternal high-fat diet on key components of the placental and hepatic endocannabinoid system

doi: 10.1152/ajpendo.00119.2017

Figure Lengend Snippet: CB1R and CB2R protein expression in the placenta from high-fat diet (HFD; n = 10) and control (CTR; n = 11) animals. A, B, E, and F: placental expression of CB1R in male fetuses (MF). C, D, G, and H: representative images of the placental sections from female fetuses (FF). Blue arrows point to syncytiotrophoblasts, green arrows point to cytotrophoblasts, and pink arrows point to fetal capillaries. The negative controls (omission of the primary antibodies) are shown as insets (A–H, top right). Scale bar, 20 μm. I–L: immunohistochemical quantifications for the HFD and CTR groups. Data are presented as means ± SE.

Article Snippet: Immunohistochemistry (IHC) was performed using a CB1R monoclonal primary antibody (cat. no. Img-CB1R-mab001; Immunogenes, Budakeszi, Hungary), a CB2R monoclonal primary antibody (cat. no. H00001269-M01; Novus Biologicals, Littleton, CO), and an anti-mouse secondary antibody (Vectastain ABC Kit anti-mouse IgG, cat. no. PK-4002; Vector Laboratories, Burlingame, CA).

Techniques: Expressing, Immunohistochemical staining

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Effect of maternal high-fat diet on key components of the placental and hepatic endocannabinoid system

doi: 10.1152/ajpendo.00119.2017

Figure Lengend Snippet: Placental expression of main endocannabinoid receptors and metabolizing enzymes in placentas of male and female fetuses near term of HFD and CTR mothers

Article Snippet: Immunohistochemistry (IHC) was performed using a CB1R monoclonal primary antibody (cat. no. Img-CB1R-mab001; Immunogenes, Budakeszi, Hungary), a CB2R monoclonal primary antibody (cat. no. H00001269-M01; Novus Biologicals, Littleton, CO), and an anti-mouse secondary antibody (Vectastain ABC Kit anti-mouse IgG, cat. no. PK-4002; Vector Laboratories, Burlingame, CA).

Techniques: Expressing

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Effect of maternal high-fat diet on key components of the placental and hepatic endocannabinoid system

doi: 10.1152/ajpendo.00119.2017

Figure Lengend Snippet: Graphic summary of presented findings in a framework of selected reported obesity-related placental changes, which are relevant to endocannabinoid transport and metabolism. Top: increased maternal and decreased fetal systemic 2-arachidonoyl glycerol (2-AG) concentrations in maternal high-fat diet (HFD) consumption. Placental 2-AG concentration is decreased in maternal obesity (MO) (13). Bottom: cross-sections of placental terminal villi. CB1R is expressed in the fetal endothelium of control (CTR) animals and additionally in the syncytiotrophoblast (ST) of the HFD group. Macrophages infiltration (17, 28), fatty acid-binding protein (FABP) and fatty acid transport protein (FATP) expression (24), lipoprotein lipase (LPL) activity (66), and lipid droplet (LD) formation (39) were increased, and docosahexaenoic acid (DHA) was reduced (16) in placentas in MO and in the HFD group. MFSD2A is expressed in placental ST and is a transporter of DHA (58, 64), which in turn is a precursor of endogenous cannabinoid (eCB) (31). Associations with eCB metabolism: LDs are functional sides of AEA inactivation (42). CB2R is expressed in the cytotrophoblast (CT) and macrophages (52), FABP is involved in eCB transport (43), and LPL activation is CB1R mediated (78). The fetal endothelium expresses monoacylglycerol lipase (MAGL), diacylglycerol lipase-α (DAGLα), N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD), and CB1R; CT expresses CB2R, DAGL, Fatty Acid Amide Hydrolase 1 (FAAH-1), NAPE-PLD, and MAGL, and ST expresses CB1R, MAGL, and FAAH-1.

Article Snippet: Immunohistochemistry (IHC) was performed using a CB1R monoclonal primary antibody (cat. no. Img-CB1R-mab001; Immunogenes, Budakeszi, Hungary), a CB2R monoclonal primary antibody (cat. no. H00001269-M01; Novus Biologicals, Littleton, CO), and an anti-mouse secondary antibody (Vectastain ABC Kit anti-mouse IgG, cat. no. PK-4002; Vector Laboratories, Burlingame, CA).

Techniques: Concentration Assay, Binding Assay, Expressing, Activity Assay, Functional Assay, Activation Assay