Journal: Neurochemical Research

Article Title: Differential Pharmacological Actions of Methadone and Buprenorphine in Human Embryonic Kidney 293 Cells Coexpressing Human μ-Opioid and Opioid Receptor-Like 1 Receptors

doi: 10.1007/s11064-011-0525-z

Figure Lengend Snippet: B max (pmol/mg protein) and K D (nM) values of μ-opioid receptors (MOR) and opioid receptor-like 1 receptors (ORL1) expressed in HEK 293 cells

Article Snippet: Non-specific binding was determined by adding 3 μM nociceptin (Tocris Bioscience, Ellisville, Missouri, USA) or DAMGO (Tocris) to the reaction mixture.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Inhibitory Effects of a Novel μ-Opioid Receptor Nonpeptide Antagonist, UD-030, on Morphine-Induced Conditioned Place Preference

doi: 10.3390/ijms24043351

Figure Lengend Snippet: ( A ) Structure of UD-030. ( B – E ) Binding properties of UD-030 for displacement of the specific binding of 1 nM [ 3 H]DAMGO, 0.7 nM [ 3 H]DADLE, 1 nM [ 3 H]U-69593, and 0.5 nM [ 3 H]nociceptin to membranes of hMOR/CHO ( B ), hDOR/CHO ( C ), hKOR/CHO ( D ), and hNOP/CHO ( E ) cells, respectively. Assays were performed on two separate occasions, each in triplicate. The data are expressed as the mean ± SEM.

Article Snippet: For the competitive binding assays, these cell membrane suspensions were stirred at 1100 rpm for 60 min on a plate shaker at room temperature with [ 3 H]DAMGO for hMOP (final concentration, 1 nM), [ 3 H]DADLE for hDOP (final concentration, 0.7 nM), [ 3 H]U-69593 for hKOP (final concentration, 1 nM), or [ 3 H]nociceptin for hNOP (final concentration, 0.5 nM) in the presence of a WGA SPA Beads (PerkinElmer, Waltham, MA, USA) suspension and various concentrations of ligands.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Inhibitory Effects of a Novel μ-Opioid Receptor Nonpeptide Antagonist, UD-030, on Morphine-Induced Conditioned Place Preference

doi: 10.3390/ijms24043351

Figure Lengend Snippet: In vitro actions of UD-030 and reference compounds against human opioid receptors and the NOP.

Article Snippet: For the competitive binding assays, these cell membrane suspensions were stirred at 1100 rpm for 60 min on a plate shaker at room temperature with [ 3 H]DAMGO for hMOP (final concentration, 1 nM), [ 3 H]DADLE for hDOP (final concentration, 0.7 nM), [ 3 H]U-69593 for hKOP (final concentration, 1 nM), or [ 3 H]nociceptin for hNOP (final concentration, 0.5 nM) in the presence of a WGA SPA Beads (PerkinElmer, Waltham, MA, USA) suspension and various concentrations of ligands.

Techniques: In Vitro, Competitive Binding Assay, Binding Assay

Journal: Cells

Article Title: Nociceptin/Orphanin FQ Opioid Peptide-Receptor Expression in the Endometriosis-Associated Nerve Fibers—Possible Treatment Option?

doi: 10.3390/cells12101395

Figure Lengend Snippet: Endometriosis patients showed increased nerve fiber and blood vessel density, as well as NOP−receptor expression. ( A ) EM and control samples stained with PGP9.5 (green) and NOP (red) antibody. Merged images showi the colocalization of pan marker (PGP9.5) and the NOP receptor. All pictures are at 200× magnification. ( B , C ) PGP9.5−positive nerve fibers per mm 2 in EM patients and controls. ( D , E ) NOP−positive nerve fibers per mm 2 in EM patients and controls. ( F ) Blood vessels per mm 2 in EM patients and controls. ( G ) NOP−positive blood vessels mm 2 in EM patients and controls. Arrows indicate colocalization points. EM: endometriosis patients; Ctr: control; H+: under hormonal treatment; H−: without hormonal treatment. All results are presented as the median, and 25th–75th percentile. Mann–Whitney and Kruskal–Wallis with Dunn’s multiple comparison tests. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections of 2 μm thickness were cut and used for immunofluorescence double staining using antibodies against NOP receptor (Santa Cruz, Heidelberg, Germany, sc-398073, 1:50 and Abcam, Cambridge, UK, ab66219, 1:380), protein gene product 9.5 (PGP 9.5—Novus, Wiesbaden Nordenstadt, Germany, NB110-58872, 1:300), substance P (SP, Santa Cruz, sc-21715, 1:500), calcitonin gene-related peptide (CGRP—Santa Cruz, sc-8857, 1:100), tyrosine hydroxylase (TH—Sigma, St. Louis, MO, USA, T2928, 1:100), and vasoactive intestinal peptide (VIP—Santa Cruz, sc-25347, 1:100).

Techniques: Expressing, Control, Staining, Marker, MANN-WHITNEY, Comparison

Journal: Cells

Article Title: Nociceptin/Orphanin FQ Opioid Peptide-Receptor Expression in the Endometriosis-Associated Nerve Fibers—Possible Treatment Option?

doi: 10.3390/cells12101395

Figure Lengend Snippet: Correlation analysis.

Article Snippet: Sections of 2 μm thickness were cut and used for immunofluorescence double staining using antibodies against NOP receptor (Santa Cruz, Heidelberg, Germany, sc-398073, 1:50 and Abcam, Cambridge, UK, ab66219, 1:380), protein gene product 9.5 (PGP 9.5—Novus, Wiesbaden Nordenstadt, Germany, NB110-58872, 1:300), substance P (SP, Santa Cruz, sc-21715, 1:500), calcitonin gene-related peptide (CGRP—Santa Cruz, sc-8857, 1:100), tyrosine hydroxylase (TH—Sigma, St. Louis, MO, USA, T2928, 1:100), and vasoactive intestinal peptide (VIP—Santa Cruz, sc-25347, 1:100).

Techniques:

Journal: Cells

Article Title: Nociceptin/Orphanin FQ Opioid Peptide-Receptor Expression in the Endometriosis-Associated Nerve Fibers—Possible Treatment Option?

doi: 10.3390/cells12101395

Figure Lengend Snippet: NOP receptors are present in sympathetic, parasympathetic, and sensory fibers innervating endometriotic lesions. ( A ) EM and control samples stained with CGRP (sensory fibers—red) and NOP (green) antibody. ( B ) EM and control samples stained with SP (sensory fibers—green) and NOP (red) antibody. ( C ) EM and control samples stained with TH (sympathetic fibers—green) and NOP (red) antibody. ( D ) EM and control samples stained with VIP (parasympathetic fibers—green) and NOP (red) antibody. Arrows indicate colocalization points between nerve marker and the NOP receptor. All pictures are 200× magnification.

Article Snippet: Sections of 2 μm thickness were cut and used for immunofluorescence double staining using antibodies against NOP receptor (Santa Cruz, Heidelberg, Germany, sc-398073, 1:50 and Abcam, Cambridge, UK, ab66219, 1:380), protein gene product 9.5 (PGP 9.5—Novus, Wiesbaden Nordenstadt, Germany, NB110-58872, 1:300), substance P (SP, Santa Cruz, sc-21715, 1:500), calcitonin gene-related peptide (CGRP—Santa Cruz, sc-8857, 1:100), tyrosine hydroxylase (TH—Sigma, St. Louis, MO, USA, T2928, 1:100), and vasoactive intestinal peptide (VIP—Santa Cruz, sc-25347, 1:100).

Techniques: Control, Staining, Marker