Journal: International Journal of Molecular Sciences

Article Title: Brexpiprazole Reduces 5-HT7 Receptor Function on Astroglial Transmission Systems

doi: 10.3390/ijms23126571

Figure Lengend Snippet: Receptor binding profiles of mood-stabilising atypical antipsychotics. Brexpiprazole (Brex), aripiprazole (APZ), clozapine (CLZ), lurasidone (LUR), quetiapine (QTP), ziprasidone (ZIP), zotepine (ZTP) against serotonin (5-HT) type 1A (5-HT1AR), type 2A (5-HT2AR), type 7 (5-HT7R) receptor, histamine type 1 (H1R) receptor and dopamine receptors type 1 (D1R) and 2 (D2R). Data are equilibrium constant (Ki) values (nM).

Article Snippet: Antibodies against GAPDH (NB300-327, 1:300, Novus Biologicals, Littleton, CO, USA), connexin43 (C6219, 1:100, Sigma-Aldrich, St. Louis, MO, USA), 5-HT1AR (NBP2-21590, 1:00, Novus Biologicals, Littleton, CO, USA), 5-HT7R (NB100-56352, 1:00, Novus Biologicals, Littleton, CO, USA), Erk (AF1576, 10 μg/mL, R&D systems, Minneapolis, MN, USA), phosphorylated Erk (AF1018, 5 μg/mL, R&D Systems), Akt (AF1775, 1 μg/mL, R&D Systems), phosphorylated Akt (AF877, 5 μg/mL, R&D Systems), AMPKα (2603, 1:50, Cell Signalling Technology, Danvers, MA, USA), and phosphorylated-AMPKα (2535, 1:50, Cell Signalling Technology, Danvers, MA, USA) were diluted in an antibody diluent (Immuno Shot Platinum, CosmoBio, Tokyo, Japan) [ ].

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Brexpiprazole Reduces 5-HT7 Receptor Function on Astroglial Transmission Systems

doi: 10.3390/ijms23126571

Figure Lengend Snippet: Effects of subchronic administration (7 and 14 days) of therapeutic relevant concentration of Brex (Brex: 300 nM) and interaction between Brex and 5-HT1A receptor (5-HT1AR) antagonist WAY100635 (WAY: 10 μM) on protein expression of 5-HT1A (panel A ) and 5-HT7 (panel B ) receptor in the plasma membrane fraction of cortical primary cultured astrocytes. In left side histograms, ordinate: mean ± SD ( n = 6) of the relative protein level of 5-HT1AR and 5-HT7R per GAPDH. * p < 0.05, ** p < 0.01: relative to control (Brex-free) by one-way analysis of variance (ANOVA) with Tukey’s post-hoc test, and @ p < 0.05: relative to Brex for 14 days by Student’s T -test. Right side panels indicate their pseudo-gel images using capillary immunoblotting.

Article Snippet: Antibodies against GAPDH (NB300-327, 1:300, Novus Biologicals, Littleton, CO, USA), connexin43 (C6219, 1:100, Sigma-Aldrich, St. Louis, MO, USA), 5-HT1AR (NBP2-21590, 1:00, Novus Biologicals, Littleton, CO, USA), 5-HT7R (NB100-56352, 1:00, Novus Biologicals, Littleton, CO, USA), Erk (AF1576, 10 μg/mL, R&D systems, Minneapolis, MN, USA), phosphorylated Erk (AF1018, 5 μg/mL, R&D Systems), Akt (AF1775, 1 μg/mL, R&D Systems), phosphorylated Akt (AF877, 5 μg/mL, R&D Systems), AMPKα (2603, 1:50, Cell Signalling Technology, Danvers, MA, USA), and phosphorylated-AMPKα (2535, 1:50, Cell Signalling Technology, Danvers, MA, USA) were diluted in an antibody diluent (Immuno Shot Platinum, CosmoBio, Tokyo, Japan) [ ].

Techniques: Concentration Assay, Expressing, Clinical Proteomics, Membrane, Cell Culture, Control, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Brexpiprazole Reduces 5-HT7 Receptor Function on Astroglial Transmission Systems

doi: 10.3390/ijms23126571

Figure Lengend Snippet: Concentration-dependent acute effects of Brex (100 and 300 nM) on intracellular cAMP level in astrocytes (panel A ). Interaction among therapeutic relevant concentration of Brex (100 and 300 nM), 5 μM AS19 (5-HT7R agonist) (panel B ) and 10 μM WAY100635 (5-HT1AR antagonist) (panel C ) on intracellular cAMP level in astrocytes. Ordinates indicate mean ± SD ( n = 6) of intracellular cAMP level in cultured astrocytes (fmol/mg). * p < 0.05, ** p < 0.01: relative to control (Brex-free) by one-way ANOVA with Tukey’s post-hoc test.

Article Snippet: Antibodies against GAPDH (NB300-327, 1:300, Novus Biologicals, Littleton, CO, USA), connexin43 (C6219, 1:100, Sigma-Aldrich, St. Louis, MO, USA), 5-HT1AR (NBP2-21590, 1:00, Novus Biologicals, Littleton, CO, USA), 5-HT7R (NB100-56352, 1:00, Novus Biologicals, Littleton, CO, USA), Erk (AF1576, 10 μg/mL, R&D systems, Minneapolis, MN, USA), phosphorylated Erk (AF1018, 5 μg/mL, R&D Systems), Akt (AF1775, 1 μg/mL, R&D Systems), phosphorylated Akt (AF877, 5 μg/mL, R&D Systems), AMPKα (2603, 1:50, Cell Signalling Technology, Danvers, MA, USA), and phosphorylated-AMPKα (2535, 1:50, Cell Signalling Technology, Danvers, MA, USA) were diluted in an antibody diluent (Immuno Shot Platinum, CosmoBio, Tokyo, Japan) [ ].

Techniques: Concentration Assay, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Brexpiprazole Reduces 5-HT7 Receptor Function on Astroglial Transmission Systems

doi: 10.3390/ijms23126571

Figure Lengend Snippet: Proposed hypothesis of the mechanisms of Brex on intracellular signalling in astrocytes. Chronic administration of a therapeutically relevant concentration of Brex downregulates both 5-HT1AR and 5-HT7R via its 5-HT1AR partial agonistic and possible 5-HT7R inverse agonistic actions, respectively. The downregulation of 5-HT1AR and 5-HT7R by Brex attenuates AMPK, Erk, and Akt signalling in astrocytes.

Article Snippet: Antibodies against GAPDH (NB300-327, 1:300, Novus Biologicals, Littleton, CO, USA), connexin43 (C6219, 1:100, Sigma-Aldrich, St. Louis, MO, USA), 5-HT1AR (NBP2-21590, 1:00, Novus Biologicals, Littleton, CO, USA), 5-HT7R (NB100-56352, 1:00, Novus Biologicals, Littleton, CO, USA), Erk (AF1576, 10 μg/mL, R&D systems, Minneapolis, MN, USA), phosphorylated Erk (AF1018, 5 μg/mL, R&D Systems), Akt (AF1775, 1 μg/mL, R&D Systems), phosphorylated Akt (AF877, 5 μg/mL, R&D Systems), AMPKα (2603, 1:50, Cell Signalling Technology, Danvers, MA, USA), and phosphorylated-AMPKα (2535, 1:50, Cell Signalling Technology, Danvers, MA, USA) were diluted in an antibody diluent (Immuno Shot Platinum, CosmoBio, Tokyo, Japan) [ ].

Techniques: Concentration Assay

Journal:

Article Title: LACK OF ANALGESIC EFFICACY OF SPINAL ONDANSETRON ON THERMAL AND MECHANICAL HYPERSENSITIVITY FOLLOWING SPINAL NERVE LIGATION IN THE RAT

doi: 10.1016/j.brainres.2010.07.020

Figure Lengend Snippet: Spinal nerve ligation (SNL) does not alter the density of serotonergic nerve fibers in the spinal cord. Photomicrographs of serotonin transporter immunoreactive (5-HTT-IR) axons at the L4 (right panels) and L5 (left panels) spinal dorsal horn of a normal rat (A,B) or 14 days after spinal nerve ligation, ipsilateral to injury (C,D). Numerous 5-HTT-IR axons are observed throughout the dorsal horn, particularly in laminae I-IIo and laminae III-V. High power photomicrographs depicting 5-HTT-IR in axonal varicosities in spinal cord dorsal horn of normal (inset b) and SNL (inset d) rats. Scale bar = 100 μm; b,d = 25 μm.

Article Snippet: Every 4 th section was incubated over night at 4°C with primary antibody for 5HT (rabbit anti-rat 5-HT, Immunostar, Minneapolis, MN) or serotonin transporter (rabbit anti-rat 5-HTT, 1:1000, Immunostar).

Techniques: Ligation