pacap antagonist pacap 1 27  (MedChemExpress)

Journal: Addiction Biology

Article Title: PACAP Signalling Network in the Nucleus Accumbens Core Regulates Reinstatement Behaviour in Rat

doi: 10.1111/adb.70090

Figure Lengend Snippet: PACAP signalling in the NAcc. (A) Western blots of NAcc punch lysates confirming the presence of PACAP receptor (PAC1R), with specificity validated using a synthetic blocking peptide. (B) Quantification of endogenous PACAP levels in the NAcc extracellular lysate by microdialysis (176.70 ± 53.27 pM, mean ± SEM). (C) Diagram illustrating retrograde tracing, highlighting the neural pathway from the prefrontal cortex (PFC) to the NAcc. (D) Confocal image (10×) of the NAc injection site showing the spread of fluorescently conjugated cholera toxin subunit B (CTb 594) labelling (magenta) and cell nuclei stained with DAPI (blue), with the anterior commissure (aca) as an anatomical landmark. (E) Brain section highlighting prefrontal cortical regions with detailed anatomical labelling, including motor areas (M1 and M2), orbital cortices (MO, VO and LO) and agranular insular regions (AIV and AID). The inset (40 × composite) shows PACAP mRNA expression (green), CTb‐labelled projection neurons (magenta), nuclear DAPI staining (blue) and a merged image highlighting colocalization (yellow arrows). Abbreviations: OB (olfactory bulb), Cm (cerebellum), PrL (prelimbic cortex), Cg1 (cingulate cortex area 1), Fr3 (frontal cortex, Area 3).

Article Snippet: Pharmacological interventions included bilateral intracranial microinfusions of vehicle (0.9% saline), PACAP 1–38 (225, 450 ng/500 nL, #350‐35, Echelon Biosciences, UT, USA), SKF81297 (3 μg/500 nL, #1447, Tocris, Bristol, UK) in 16% DMSO or sumanirole (100 ng/500 nL, #2773, Tocris, Bristol) at 125 nL/min in 16% DMSO 10 min before testing, with lever pressing recorded for 2 h. All DMSO solutions were prepared in 0.9% saline.

Techniques: Western Blot, Blocking Assay, Retrograde Tracing, Injection, Staining, Expressing

Journal: Addiction Biology

Article Title: PACAP Signalling Network in the Nucleus Accumbens Core Regulates Reinstatement Behaviour in Rat

doi: 10.1111/adb.70090

Figure Lengend Snippet: PACAP 1–38 microinjection in the NAcc does not induce cocaine‐seeking behaviour. (A) Active lever presses (mean ± SEM) during a 2‐h session in extinction‐trained rats compared to those tested following intra‐NAcc microinjection of PACAP 1–38 (450 ng/500 nL). Lever pressing did not significantly differ between the test sessions, indicating that PACAP 1–38 administered into the NAcc at this dose is not sufficient to produce cocaine‐seeking behaviour ( n = 16/condition). (B) Schematic illustration of PACAP 1–38 microinjection sites within the NAcc, confirmed by histological analysis.

Article Snippet: Pharmacological interventions included bilateral intracranial microinfusions of vehicle (0.9% saline), PACAP 1–38 (225, 450 ng/500 nL, #350‐35, Echelon Biosciences, UT, USA), SKF81297 (3 μg/500 nL, #1447, Tocris, Bristol, UK) in 16% DMSO or sumanirole (100 ng/500 nL, #2773, Tocris, Bristol) at 125 nL/min in 16% DMSO 10 min before testing, with lever pressing recorded for 2 h. All DMSO solutions were prepared in 0.9% saline.

Techniques: Microinjection

Journal: Addiction Biology

Article Title: PACAP Signalling Network in the Nucleus Accumbens Core Regulates Reinstatement Behaviour in Rat

doi: 10.1111/adb.70090

Figure Lengend Snippet: PACAP 1–38 microinjection in the NAcc inhibits cocaine‐primed reinstatement. (A) Lever presses (mean ± SEM) during a 2‐h session by extinguished cocaine‐seeking rats reinstated with an intraperitoneal injection of 10‐mg/kg cocaine. PACAP 1–38 microinjected into the NAcc 10 min prior to the session significantly reduced cocaine‐seeking behaviour in a dose‐dependent manner ( n = 6–11). * and # indicate Holm–Sidak, p < 0.05 compared to the extinction and vehicle groups, respectively. (B) Schematic showing the injection sites within the NAcc.

Article Snippet: Pharmacological interventions included bilateral intracranial microinfusions of vehicle (0.9% saline), PACAP 1–38 (225, 450 ng/500 nL, #350‐35, Echelon Biosciences, UT, USA), SKF81297 (3 μg/500 nL, #1447, Tocris, Bristol, UK) in 16% DMSO or sumanirole (100 ng/500 nL, #2773, Tocris, Bristol) at 125 nL/min in 16% DMSO 10 min before testing, with lever pressing recorded for 2 h. All DMSO solutions were prepared in 0.9% saline.

Techniques: Microinjection, Injection

Journal: Addiction Biology

Article Title: PACAP Signalling Network in the Nucleus Accumbens Core Regulates Reinstatement Behaviour in Rat

doi: 10.1111/adb.70090

Figure Lengend Snippet: PACAP 1–38 attenuates D1‐like dopamine receptor agonist‐primed reinstatement. (A) Lever presses (mean ± SEM) during a 2‐h session by extinguished rats following intra‐NAcc administration of the D1‐like dopamine receptor agonist SKF81297 (3 μg/500 nL). SKF81297 ‐induced reinstatement of cocaine‐seeking behaviour was significantly attenuated by PACAP 1–38 (450 ng/500 nL; n = 10–12). * and # indicate Holm–Sidak, p < 0.05 compared to the extinction and vehicle groups, respectively. (B) Schematic showing the injection sites for SKF81297 and PACAP 1–38 within the NAcc.

Article Snippet: Pharmacological interventions included bilateral intracranial microinfusions of vehicle (0.9% saline), PACAP 1–38 (225, 450 ng/500 nL, #350‐35, Echelon Biosciences, UT, USA), SKF81297 (3 μg/500 nL, #1447, Tocris, Bristol, UK) in 16% DMSO or sumanirole (100 ng/500 nL, #2773, Tocris, Bristol) at 125 nL/min in 16% DMSO 10 min before testing, with lever pressing recorded for 2 h. All DMSO solutions were prepared in 0.9% saline.

Techniques: Injection

Journal: Addiction Biology

Article Title: PACAP Signalling Network in the Nucleus Accumbens Core Regulates Reinstatement Behaviour in Rat

doi: 10.1111/adb.70090

Figure Lengend Snippet: PACAP 1–38 has no effect on D2‐like dopamine receptor agonist‐primed reinstatement in drug seeking. (A) The data (mean ± SEM) show that administration of the D2‐like dopamine receptor agonist sumanirole (100 ng/500 nL) moderately induces reinstatement in cocaine‐experienced rats and PACAP 1–38 (450 ng/500 nL) coadministration does not inhibit it ( n = 11–12/group). * indicates Holm–Sidak, p < 0.05 compared to the extinction. (B) Schematic illustrates the sumanirole and PACAP injection sites.

Article Snippet: Pharmacological interventions included bilateral intracranial microinfusions of vehicle (0.9% saline), PACAP 1–38 (225, 450 ng/500 nL, #350‐35, Echelon Biosciences, UT, USA), SKF81297 (3 μg/500 nL, #1447, Tocris, Bristol, UK) in 16% DMSO or sumanirole (100 ng/500 nL, #2773, Tocris, Bristol) at 125 nL/min in 16% DMSO 10 min before testing, with lever pressing recorded for 2 h. All DMSO solutions were prepared in 0.9% saline.

Techniques: Injection

Journal: Addiction Biology

Article Title: PACAP Signalling Network in the Nucleus Accumbens Core Regulates Reinstatement Behaviour in Rat

doi: 10.1111/adb.70090

Figure Lengend Snippet: PACAP signalling in the NAcc. (A) Western blots of NAcc punch lysates confirming the presence of PACAP receptor (PAC1R), with specificity validated using a synthetic blocking peptide. (B) Quantification of endogenous PACAP levels in the NAcc extracellular lysate by microdialysis (176.70 ± 53.27 pM, mean ± SEM). (C) Diagram illustrating retrograde tracing, highlighting the neural pathway from the prefrontal cortex (PFC) to the NAcc. (D) Confocal image (10×) of the NAc injection site showing the spread of fluorescently conjugated cholera toxin subunit B (CTb 594) labelling (magenta) and cell nuclei stained with DAPI (blue), with the anterior commissure (aca) as an anatomical landmark. (E) Brain section highlighting prefrontal cortical regions with detailed anatomical labelling, including motor areas (M1 and M2), orbital cortices (MO, VO and LO) and agranular insular regions (AIV and AID). The inset (40 × composite) shows PACAP mRNA expression (green), CTb‐labelled projection neurons (magenta), nuclear DAPI staining (blue) and a merged image highlighting colocalization (yellow arrows). Abbreviations: OB (olfactory bulb), Cm (cerebellum), PrL (prelimbic cortex), Cg1 (cingulate cortex area 1), Fr3 (frontal cortex, Area 3).

Article Snippet: Antibody specificity was confirmed by reprobing with and without a synthetic blocking PAC1R peptide (1:600, #BLP‐VR003, Alomone Labs, Jerusalem, Israel).

Techniques: Western Blot, Blocking Assay, Retrograde Tracing, Injection, Staining, Expressing