Journal: Physiological Reports

Article Title: Ontogeny and programming of the fetal temporal cortical endocannabinoid system by moderate maternal nutrient reduction in baboons ( Papio spp.)

doi: 10.14814/phy2.14024

Figure Lengend Snippet: Results of the ANOVA for treatment (TRT; CTR vs. MNR), gestational age (GA; 120 vs. 140 vs. 165 dGA) and the interaction between treatment and age (TRT × GA) for CB1R and CB2R gene expression in male and female fetuses

Article Snippet: The membranes were blocked with 5% fat‐free milk (Cat. # 170‐6404, Bio‐Rad, USA) for 1 h. The membranes were then probed with primary antibodies to detect CB1R (1:1000; Cat. # IMG‐CB1R‐mAb001 Immunogenes, USA), CB2R (1:2000; Cat. # H00001269‐M01, Abnova, Taiwan), FAAH‐1 (1:1000; Cat. # ab54615, Abcam, USA), DAGL α (1:1000; Cat. # SC‐390409, Santa Cruz Biotechnology Inc., USA), MAGL (1:1000; Cat. # 100035, Cayman Chemical, Ann Arbor, MI, USA), COX‐2 (1:1000; Cat. # ab62331, Abcam, Cambridge, MA, USA), and NAPE‐PLD (1:1000; Cat. # 10305, Cayman Chemical, USA) in 1% BSA (Cat. # A4503, Sigma‐Aldrich) in TBS‐T.

Techniques: Expressing

Journal: Physiological Reports

Article Title: Ontogeny and programming of the fetal temporal cortical endocannabinoid system by moderate maternal nutrient reduction in baboons ( Papio spp.)

doi: 10.14814/phy2.14024

Figure Lengend Snippet: Results of ANOVA for treatment (TRT; CTR vs. MNR), gestational age (GA; 120 vs. 140 vs. 165 dGA) and the interaction between treatment and age (TRT × GA) for protein expression in male and female fetuses

Article Snippet: The membranes were blocked with 5% fat‐free milk (Cat. # 170‐6404, Bio‐Rad, USA) for 1 h. The membranes were then probed with primary antibodies to detect CB1R (1:1000; Cat. # IMG‐CB1R‐mAb001 Immunogenes, USA), CB2R (1:2000; Cat. # H00001269‐M01, Abnova, Taiwan), FAAH‐1 (1:1000; Cat. # ab54615, Abcam, USA), DAGL α (1:1000; Cat. # SC‐390409, Santa Cruz Biotechnology Inc., USA), MAGL (1:1000; Cat. # 100035, Cayman Chemical, Ann Arbor, MI, USA), COX‐2 (1:1000; Cat. # ab62331, Abcam, Cambridge, MA, USA), and NAPE‐PLD (1:1000; Cat. # 10305, Cayman Chemical, USA) in 1% BSA (Cat. # A4503, Sigma‐Aldrich) in TBS‐T.

Techniques: Expressing

Journal: Frontiers in Molecular Neuroscience

Article Title: Spinal cannabinoid receptor 2 activation alleviates neuropathic pain by regulating microglia and suppressing P2X7 receptor

doi: 10.3389/fnmol.2023.1061220

Figure Lengend Snippet: SNI induced hyperalgesia changed the expression and morphology of microglia in the spinal dorsal horn. (A,B) SNI significantly reduced the MWT (A) and improved the acetone score (B) in the ipsilateral paws. (**** p < 0.0001 compared with the sham group, n = 12 in each group). (C) Double immunofluorescence of CB2R and Iba1 in the spinal cord of sham and SNI-7d rats. The fluorescence images in the boxes were magnified to better display the double staining. Scale bar, 100 μm. (D) The expression of Iba1 increased on days 3, 7, and 14 after SNI. (* p < 0.05 compared with the sham group, n = 6 in each group). (E) microglia micrographs from the ipsilateral spinal dorsal horn sham or SNI-7d rats. The images with higher magnification were skeletonized and expressed as black-and-white images (right side) for measuring the three parameters. Scale bars, 100 μm for lower magnification and 50 μm for higher magnification. (F) Quantification of the area of Iba1+ cells and morphology of microglia. Solidity, the area of cell body / the area of processes for each cell. Results for each group were obtained from three rats and three micrographs from each sample. The average values of each sample were taken for statistics. (** p < 0.01, *** p < 0.001 compared with the sham group).

Article Snippet: The CB2R agonist PM226 (GC50387, GLPBIO, United States) was dissolved in normal saline.

Techniques: Expressing, Immunofluorescence, Fluorescence, Double Staining

Journal: Frontiers in Molecular Neuroscience

Article Title: Spinal cannabinoid receptor 2 activation alleviates neuropathic pain by regulating microglia and suppressing P2X7 receptor

doi: 10.3389/fnmol.2023.1061220

Figure Lengend Snippet: The mechanical and cold hyperalgesia were alleviated in the ipsilateral paws of rats after PM226 treatment. (A) Timeline of the experimental process. (B,D) Continuous intrathecal administration of PM226 improved the MWT (B) and decreased the acetone score (D) in the ipsilateral paws (C,E) . There was no significant difference in the MWT and acetone score between the two groups after PM226 treatment (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 compared with SNI + Vehicle group, n = 10 in each group). MWT, mechanical withdrawal threshold; AT, acetone test.

Article Snippet: The CB2R agonist PM226 (GC50387, GLPBIO, United States) was dissolved in normal saline.

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Spinal cannabinoid receptor 2 activation alleviates neuropathic pain by regulating microglia and suppressing P2X7 receptor

doi: 10.3389/fnmol.2023.1061220

Figure Lengend Snippet: PM226 blocks the proinflammatory process and promotes microglia to be beneficial. (A) Western blot showed that the expression of TNF-α and CD86 was significantly decreased in the spinal dorsal cord after intrathecal administration for 7 days. (B) The expression of Arg-1 and CD163 was significantly increased in the spinal dorsal cord after intrathecal administration for 7 days. (* p < 0.05, ** p < 0.01, *** p < 0.001 compared with SNI + Vehicle group, n = 6 in each group). (C) Double immunostaining of CD86 (green) and Iba1 (red) in the spinal dorsal horn. Arrows showed the representative merge of CD86 and Iba1. (D) Double immunostaining of CD163 (green) and Iba1 (red) in the spinal dorsal horn. Arrows showed the representative merge of CD163 and Iba1. Scale bars, 100 μm for lower magnification and 50 μm for higher magnification. (E) Histograms are quantifications of the percentage of CD86 + −Iba1 + cells and CD163 + −Iba1 + cells. Results for each group were obtained from three rats, and three micrographs from each sample. The average values of each sample were taken for statistics. (** p < 0.01 compared with SNI + Vehicle group). (F) The expression of TNF-α mRNA, CD86, and iNOS mRNA was significantly decreased and Arg-1, CD163, and IL-10 mRNA was increased in the spinal dorsal cords after intrathecal administration of PM226 for 7 days (* p < 0.05, ** p < 0.01, *** p < 0.001 compared with SNI + Vehicle group, n = 4 in each group). S + V, SNI + Vehicle group; S + P, SNI + PM226 group.

Article Snippet: The CB2R agonist PM226 (GC50387, GLPBIO, United States) was dissolved in normal saline.

Techniques: Western Blot, Expressing, Double Immunostaining

Journal: Frontiers in Molecular Neuroscience

Article Title: Spinal cannabinoid receptor 2 activation alleviates neuropathic pain by regulating microglia and suppressing P2X7 receptor

doi: 10.3389/fnmol.2023.1061220

Figure Lengend Snippet: PM226 suppressed P2X7 in the rat spinal dorsal horn. (A) Western blot showed that the expression of P2X7 increased with time, and significantly increased on days 7 and 14 after SNI. (* p < 0.05, ** p < 0.01 compared with sham group, n = 6 in each group). (B) The mean fluorescence intensity of P2X7 ( C , red) in the ipsilateral spinal dorsal horn of the sham and SNI 7d group. Results for each group were obtained from three rats, and three micrographs from each sample. The average values of each sample were taken for statistics. (* p < 0.05, compared with sham group). (C) Double immunostaining of P2X7 (red) and Iba1 (green) in the dorsal horn of the spinal cord. Arrows showed the representative merge of P2X7 and Iba1. Scale bars, 100 μm for lower magnification and 50 μm for higher magnification. (D) The administration of A-438079 on day 3 improved the MWT of the ipsilateral paws on days 5 and 7 (* p < 0.05 compared FIGURE 5 (Continued)with SNI + Vehicle group, n = 8 in each group). (E) The IBA1 protein level and P2X7 protein level in the spinal cord both decreased after A-438079 administration (** p < 0.01 compared with SNI + Vehicle group, n = 6 in each group). (F) The mean fluorescence intensity of Iba1 of ipsilateral spinal dorsal horn in the SNI + A-438079 group was significantly lower than that in the SNI + Vehicle group. Results of each group were obtained from three rats, and three micrographs from each sample. The average values of each sample were taken for statistics (* p < 0.05 compared with SNI + Vehicle group). (G) The protein expression of P2X7 was significantly decreased in the spinal dorsal cord after intrathecal administration of PM226 for 7 days. (*** p < 0.001 compared with SNI + Vehicle group, n = 6 in each group). (H) The mean fluorescence intensity of P2X7 ( F , red) in the ipsilateral spinal dorsal horn after intrathecal administration for 7 days. Results of each group were obtained from three rats, and three micrographs from each sample. The average values of each sample were taken for statistics. (* p < 0.05 compared with SNI + Vehicle group). (I) Double immunostaining of P2X7 (red) and Iba1 (green) in the spinal dorsal horn after intrathecal administration. Arrows showed the representative merge of P2X7 and Iba1. Scale bars, 100 μm for lower magnification and 50 μm for higher magnification.

Article Snippet: The CB2R agonist PM226 (GC50387, GLPBIO, United States) was dissolved in normal saline.

Techniques: Western Blot, Expressing, Fluorescence, Double Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Structure of the CB2 receptor. ( A ) Side view. ( B ) Top view. The transmembrane helices 1, 2, 3, 4, 5, 6, and 7 are shown in blue, green, light pink, orange, pink, yellow and red colors, respectively. The intracellular and extracellular loops are shown in black, and the extracellular N terminus, helix 8, and the intracellular C terminus are shown in gray. A representative ligand in the orthosteric binding site is rendered in tan color spheres.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Binding affinity of THC to CB2R with and without vitamin E acetate.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Hydrogen bonds between vitamin Es and ( A ) the active state structure of CB2R, ( B ) the inactive state structure of CB2R, ( C ) the phospholipid bilayer surrounding the active state structure of CB2R, and ( D ) the phospholipid bilayer surrounding the inactive state structure of CB2R.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: The average positions of vitamin Es in the membrane of active state ( left panel ) and inactive state structures ( right panel ) of CB2R. The average positions of phosphates in the upper and lower layers are shown as orange lines, and the positions of the five vitamin Es as black, blue, magenta, green, and violet lines.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques: Membrane

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Contact frequencies of the amino acid residues of the active state structure of CB2R with α (black), β (red), γ (green) and δ (blue) vitamin Es.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Contact frequencies of the amino acid residues of the inactive state structure of CB2R with α (black), β (red), γ (green) and δ (blue) vitamin Es.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Interaction fractions (upper panel) and 2D interaction diagram (lower panel) of THC with active state structure of CB2R in ( A ) the absence of α vitamin E and its acetate, ( B ) in the presence of α vitamin Es, and ( C ) in the presence of α vitamin E acetate.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Last frames of ( A ) system 1 and ( B ) system 2. THC and Ser 285 are shown in green, CB2R in gray, and water in blue.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Potential Pro-Inflammatory Effect of Vitamin E Analogs through Mitigation of Tetrahydrocannabinol (THC) Binding to the Cannabinoid 2 Receptor

doi: 10.3390/ijms23084291

Figure Lengend Snippet: Different systems and time scales for MD simulation.

Article Snippet: The crystal structure of inactive state CB2R (PDB 5ZTY) was obtained from the protein databank [ ].

Techniques: