Diabète, Insuline & Glucose - page 1 - Publications Scientifiques

Dernière publication 01/05/2026

Divergence in μ and δ opioid receptor pharmacology in neurons and antinociceptiv

Calcium imaging experiments were performed 24–36 h after seeding. Cells were incubated in a solution containing 2 μM Fura-2 acetoxymethyl ester...

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    [title] => Divergence in μ and δ opioid receptor pharmacology in neurons and antinociceptiv
    [paragraph] => Divergence in μ and δ opioid receptor pharmacology in neurons and antinociceptive efficacy in neuropathic pain- Insights from MP135 and CYM51010
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Authors
Perrine Inquimbert, Chantal Fitterer, Sylvain Hugel, Mila Jesic, Yannick Goumon, Virgine Andry, Severine Schneider, Francois Daubeuf, Abdelfattah Faouzi, Stephane Doridot, Susruta Majumdar, Frederic Bihel, Martine Schmitt, Dominique Massotte

 Lab
Centre National de la Recherche Scientifique; Interdisciplinary Thematic Institute IMS
Journal
British Journal of Pharmacology
Abstract
Calcium imaging experiments were performed 24–36 h after seeding. Cells were incubated in a solution containing 2 μM Fura-2 acetoxymethyl ester (F1201, Molecular Probes, Eugene, USA) and 0.01% pluronic acid (Molecular Probes, Eugene, USA) for 1 h at room temperature. During intracellular Ca2+measurements, cells were continuously perfused with an extracellular solution (130 mM NaCl; 5 mM KCl; 1 mM CaCl2; 1 mM MgCl2; 10 mM glucose; 10 mM HEPES pH 7.3) maintained at 34°C. Fluorescence measurements were performed on an inverted microscope (Axiovert 35, Zeiss, Rueil Malmaison, France) with a 40X oil-immersion objective (Fluo 40, NA 1.30, Nikon, Amstelveen, The Netherlands) using a real-time imaging system with a cooled CCD camera (CoolSnap HQ, Photometrics) and an imaging analysis software (Imaging Workbench software 4.0, Axon Instruments, Union City, USA). Fluorescence was alternatively excited at 340 and 380 nm with a Lambda-10 filter wheel (Sutter Instruments, Novato, USA), and the emitted light was collected above 520 nm. Images were acquired every 2 s and intracellular Ca2+concentration was expressed using the fluorescence ratio F340/F380 after background subtraction. Data are presented as mean ± SEM from five independent cultures per condition (WT, δ KO, μ KO), each prepared from at least two animals, corresponding to a total of at least 10 animals per condition. Each drug was applied only once per neuron with WT morphine n = 6 neurons/culture, WT CYM51010 n = 9 neurons/culture, WT MP135 n = 11 neurons/culture, δ KO morphine n = 34 neurons/culture, δ KO CYM51010 n = 27 neurons/culture, δ KO MP135 n = 23 neurons/culture,μKO morphine n = 9 neurons/culture,μKO CYM51010 n = 12 neurons/culture andμKO MP135 n = 13 neurons/culture. The effects of morphine (1 μM), CYM51010 (400 nM) and MP135 (100 nM) were assessed by measuring changes in intracellular Ca2+concentration in response to 50 mM KCl. For each neuron, responses to KCl alone were compared with those obtained when KCl was co-applied with the drug. Drug effects were quantified as the ratio of the amplitude of the KCl + drug response to the amplitude of the KCl alone response, with each neuron serving as its own control. All cells exhibiting a detectable response to 50 mM KCl were included in the analysis. Neuronal surface area was measured using ImageJ software, and the average cell diameter was estimated according to diameter = 2√(surface/π). Only morphologically isolated neurons were included in surface measurements.
Keywords/Topics
G protein-coupled receptor; heteromer; internalization; neuropathic pain; opioid receptor; respiratory depression; sex difference; signalling
BIOSEB Instruments Used:
Von Frey Filaments (BIO-VF-M)
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