ness 0327  (Bio-Techne corporation)

Journal: Research

Article Title: Endocannabinoids Block Headache and Anxiety Comorbidity via Two-Pronged Anterior Insular Projections

doi: 10.34133/research.1031

Figure Lengend Snippet: CB1R antagonism in PrL- and ovBNST-reversed JZL184 produced analgesic and anxiolytic effects in mice with repeated ISDN injections. (A to C and K to M) Experimental schedule (A and K) and schematic diagram (B and L) for bilateral cannula implantation above bilateral PrLs or ovBNSTs, local injection of the CB1R antagonist NESS0327 into bilateral PrLs (B and C) or ovBNSTs (L and M), and intraperitoneal injection of 2-AG hydrolase (MAGL) inhibitor JZL184. Representative images show cannula tracks above PrLs (C) and ovBNSTs (M). The green fluorescent CTB-488 dye enables visualization of injection sites. Scale bars, 200 μm. (D to J and N to T) Effects of intra-PrL (D to J) or intra-ovBNST injection (N to T) of NESS0327 (0.5 μM/300 nl/side) and systematic administration of JZL184 (10 mg/kg) on repeated ISDN-induced cephalic cutaneous allodynia in Von Frey test (D and N) or anxiety-like behaviors in the OFT (E to G and O to Q) and EPM tests (H to J and R to T). The data are presented as the mean ± SEM. * P <0.05, ** P <0.01 versus Vehicle group. Detailed statistical results are provided in Table .

Article Snippet: For pharmacological inhibition of the CB1R in PrL or ovBNST, 300 nl/side of NESS0327 (0.5 μM, HY-117139, MedChemExpress, USA) was intracerebrally perfused into PrL or ovBNST.

Techniques: Produced, Injection

Journal: Frontiers in Neuroscience

Article Title: Endocannabinoid-mediated regulation of depression in the ovBNST

doi: 10.3389/fnins.2025.1629351

Figure Lengend Snippet: The cnr1 mRNA is abundantly expressed in the mPFC-ovBNST projection and acute stress increases endocannabinoid release in the ovBNST. (A) Schematic showing the retrograde tracing of upstream nuclei with AAV-Retro Plus-hSyn-Cre-EGFP infusion in the ovBNST. (B1,B2) Representative overview ( B1 ) and enlarged view ( B2 ) illustrating the injection site in the ovBNST. (C1–C8) Representative images showing retrogradely labeled neurons distributed in the upstream nuclei including the mPFC (C1) , ACC (C2) , AIP (C3) , PVA (C4) , PVT (C5) , BLA (C6) , BMP (C6) , BLP (C7) , AHiPM (C7) , PMCO (C7) , and APir (C8) . (D) Illustrative representation of cFos staining in the mPFC. (E,F) Quantification of cFos cells in the mPFC of mice exposed to the AFS ( E , n = 7) and ARS ( F , n = 6). (G) Representative images of cnr1 positive mPFC neurons projecting to the ovBNST. (H) Pie chart depicting the percentage of cnr1 + /EGFP + cells in the ovBNST ( n = 5). (I,J) Schematic (I) and representative (J) images showing the injection of AAV-hSyn-eCB2.0 into the mPFC and implantation of recording fiber above the ovBNST. (K) Representative traces of the eCB2.0 fluorescence in the ovBNST of naïve mice in responding to AFS. (L) Bar graphs showing quantitative statistics of the endocannabinoid signals ΔF/F (%) Peak ( n = 5). Data are presented as the mean ± SEM; *P < 0.05, ***P < 0.001, ****P < 0.0001, two-tailed unpaired or paired Student’s t -test. Scale bars: 200 μm for overviews and 100 μm or 50 μm for enlarged views.

Article Snippet: The CB1R antagonist NESS0327 (NESS, Cat# HY-117139), the CB1R antagonist AM251 (Cat# HY-15443), and the MAGL inhibitor JZL-184 (Cat# HY-15249) were purchased from MedChem Express.

Techniques: Retrograde Tracing, Injection, Labeling, Staining, Fluorescence, Two Tailed Test

Journal: Frontiers in Neuroscience

Article Title: Endocannabinoid-mediated regulation of depression in the ovBNST

doi: 10.3389/fnins.2025.1629351

Figure Lengend Snippet: NESS-induced CB1R blockade in the ovBNST reduces sucrose preference and increases immobility in the FST. (A) Experimental diagram illustrating the pharmacological manipulation of CB1R and behavioral tests. (B) Representative images showing bilateral implantation of cannula above the ovBNST. (C,D) Analysis of sucrose preference (%) in the SPT (C) and immobile duration in the FST (D) following NESS infusion into the ovBNST. (E,F) Representative track plots ( E ) and time in center (F) in the OFT of the mice receiving infusion of vehicle or NESS into the ovBNST. Data are presented as the mean ± SEM; *P < 0.05, two-tailed unpaired Student’s t -test. Scale bar: 500 μm.

Article Snippet: The CB1R antagonist NESS0327 (NESS, Cat# HY-117139), the CB1R antagonist AM251 (Cat# HY-15443), and the MAGL inhibitor JZL-184 (Cat# HY-15249) were purchased from MedChem Express.

Techniques: Two Tailed Test

Journal: Frontiers in Neuroscience

Article Title: Endocannabinoid-mediated regulation of depression in the ovBNST

doi: 10.3389/fnins.2025.1629351

Figure Lengend Snippet: Increased endocannabinoid signaling in the ovBNST ameliorates CUMS induced depressive-like behaviors. (A) Experimental diagram illustrating the CUMS modeling and pharmacological manipulation of CB1R with the CB1R agonist WIN or MAGL inhibitor JZL-184 administration in the ovBNST. (B) Representative images showing bilateral implantation of cannula above the ovBNST. (C–G) Analysis of sucrose preference (%) in the SPT (C) , immobile duration in the FST (D) and TST (E) , total distance travelled (F) and time in center (G) in the OFT following the CB1R agonist WIN infusion into the ovBNST in CTRL and CUMS mice. (H–L) Data corresponding to (C–G) , but for the mice subjected to the MAGL inhibitor JZL-184 infusion. Data are presented as the mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, data were analyzed using one-way ANOVA followed by Tukey-Kramer post hoc test for pairwise comparisons. Scale bar: 500 μm.

Article Snippet: The CB1R antagonist NESS0327 (NESS, Cat# HY-117139), the CB1R antagonist AM251 (Cat# HY-15443), and the MAGL inhibitor JZL-184 (Cat# HY-15249) were purchased from MedChem Express.

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Endocannabinoid-mediated regulation of depression in the ovBNST

doi: 10.3389/fnins.2025.1629351

Figure Lengend Snippet: Alterations in cnr1 mRNA and CB1R protein levels in the mPFC-ovBNST pathway of CUMS mice. (A) Representative images showing alterations in cnr1 mRNA levels in the mPFC following CUMS exposure. (B) Quantification of cnr1 mRNA levels in the mPFC of CTRL and CUMS mice ( n = 7). (C) Representative images showing alterations in cnr1 mRNA levels in the ovBNST following CUMS exposure. (D) Quantification of cnr1 mRNA levels in the ovBNST of CTRL and CUMS mice ( n = 6). (E–H) Representative images showing alterations in CB1R protein levels in the mPFC following CUMS exposure and quantification of CB1R protein levels in the ovBNST (E) , mPFC (F) , ACC (G) , and BLA (H) in CTRL and CUMS mice (ovBNST, CTRL, n = 15, CUMS, n = 12; mPFC, CTRL, n = 11, CUMS, n = 9; ACC, CTRL, n = 11, CUMS, n = 9; BLA, CTRL, n = 11, CUMS, n = 8). Data are presented as the mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed unpaired Student’s t -test. Scale bar: 200 μm.

Article Snippet: The CB1R antagonist NESS0327 (NESS, Cat# HY-117139), the CB1R antagonist AM251 (Cat# HY-15443), and the MAGL inhibitor JZL-184 (Cat# HY-15249) were purchased from MedChem Express.

Techniques: Two Tailed Test