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Neuropeptide FF (NPFF) analogs functionally antagonize opioid activities in NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells.
Mol Pharmacol. 2005 Mar; 67(3):965-75.MP

Abstract

To elucidate the mechanism of the cellular antiopioid activity of neuropeptide FF (NPFF), we have transfected the SH-SY5Y neuroblastoma cell line, which expresses mu-and delta-opioid receptors, with the human NPFF2receptor. The selected clone, SH2-D9, expressed high-affinity NPFF2 receptors in the same range order as mu- and delta-opioid receptors (100-300 fmol/mg of protein). The NPFF analog [D-Tyr1, (NMe)Phe3]NPFF (1DMe) did not modify the binding parameters of the mu- and delta-specific agonists [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin and deltorphin-I, respectively. 1DMe dose dependently inhibited 75 to 80% of the cAMP production stimulated by forskolin. Preincubation with 1DMe halved the maximal inhibition of N-type Ca2+ channels by opioid agonists. In the presence of carbachol, acting on muscarinic receptors to release Ca2+ from the intracellular stores, deltorphin-I and 1DMe enhanced this release. Preincubation with 1DMe reduced the maximal effect of deltorphin-I by 40%, demonstrating an antiopioid effect in this experimental model for the first time. By using peptides corresponding to the carboxyl terminus of the alphai1,2, alphai3, alphao, and alphas subunits of G proteins, which specifically uncouple receptors from G proteins, we demonstrated that mu-opioid and NPFF2 receptors couple to the four subunits assayed. The Ca2+ release from the intracellular stores by 1DMe resulted from the coupling of NPFF2 receptors with Galphao and Galphai1,2, whereas the coupling with Galphas reduced the antiopioid effect of 1DMe in the modulation of N-type channels. This SH2-D9 cell line now provides the opportunity to study the interaction between both receptors.

Authors+Show Affiliations

Institut de Pharmacologie et de Biologie Structurale, CNRS, UMR 5089, 205 route de Narbonne, 31077 Toulouse cedex 04, France.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15608144

Citation

Mollereau, Catherine, et al. "Neuropeptide FF (NPFF) Analogs Functionally Antagonize Opioid Activities in NPFF2 Receptor-transfected SH-SY5Y Neuroblastoma Cells." Molecular Pharmacology, vol. 67, no. 3, 2005, pp. 965-75.
Mollereau C, Mazarguil H, Zajac JM, et al. Neuropeptide FF (NPFF) analogs functionally antagonize opioid activities in NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells. Mol Pharmacol. 2005;67(3):965-75.
Mollereau, C., Mazarguil, H., Zajac, J. M., & Roumy, M. (2005). Neuropeptide FF (NPFF) analogs functionally antagonize opioid activities in NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells. Molecular Pharmacology, 67(3), 965-75.
Mollereau C, et al. Neuropeptide FF (NPFF) Analogs Functionally Antagonize Opioid Activities in NPFF2 Receptor-transfected SH-SY5Y Neuroblastoma Cells. Mol Pharmacol. 2005;67(3):965-75. PubMed PMID: 15608144.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Neuropeptide FF (NPFF) analogs functionally antagonize opioid activities in NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells. AU - Mollereau,Catherine, AU - Mazarguil,Honoré, AU - Zajac,Jean-Marie, AU - Roumy,Michel, Y1 - 2004/12/17/ PY - 2004/12/21/pubmed PY - 2005/4/13/medline PY - 2004/12/21/entrez SP - 965 EP - 75 JF - Molecular pharmacology JO - Mol Pharmacol VL - 67 IS - 3 N2 - To elucidate the mechanism of the cellular antiopioid activity of neuropeptide FF (NPFF), we have transfected the SH-SY5Y neuroblastoma cell line, which expresses mu-and delta-opioid receptors, with the human NPFF2receptor. The selected clone, SH2-D9, expressed high-affinity NPFF2 receptors in the same range order as mu- and delta-opioid receptors (100-300 fmol/mg of protein). The NPFF analog [D-Tyr1, (NMe)Phe3]NPFF (1DMe) did not modify the binding parameters of the mu- and delta-specific agonists [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin and deltorphin-I, respectively. 1DMe dose dependently inhibited 75 to 80% of the cAMP production stimulated by forskolin. Preincubation with 1DMe halved the maximal inhibition of N-type Ca2+ channels by opioid agonists. In the presence of carbachol, acting on muscarinic receptors to release Ca2+ from the intracellular stores, deltorphin-I and 1DMe enhanced this release. Preincubation with 1DMe reduced the maximal effect of deltorphin-I by 40%, demonstrating an antiopioid effect in this experimental model for the first time. By using peptides corresponding to the carboxyl terminus of the alphai1,2, alphai3, alphao, and alphas subunits of G proteins, which specifically uncouple receptors from G proteins, we demonstrated that mu-opioid and NPFF2 receptors couple to the four subunits assayed. The Ca2+ release from the intracellular stores by 1DMe resulted from the coupling of NPFF2 receptors with Galphao and Galphai1,2, whereas the coupling with Galphas reduced the antiopioid effect of 1DMe in the modulation of N-type channels. This SH2-D9 cell line now provides the opportunity to study the interaction between both receptors. SN - 0026-895X UR - https://www.unboundmedicine.com/medline/citation/15608144/Neuropeptide_FF__NPFF__analogs_functionally_antagonize_opioid_activities_in_NPFF2_receptor_transfected_SH_SY5Y_neuroblastoma_cells_ L2 - http://molpharm.aspetjournals.org/cgi/pmidlookup?view=long&pmid=15608144 DB - PRIME DP - Unbound Medicine ER -