Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors

Valerie Audinot, Anne Bonnaud, Line Grandcolas, Marianne Rodriguez, Nadine Nagel, Jean-Pierre Galizzi, Ales Balik, Sophie Messager, David G. Hazlerigg, Perry Barrett, Philippe Delagrange, Jean A. Boutin

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

In order to interpret the effects of melatonin ligands in rats, we need to determine their activity at the receptor subtype level in the corresponding species. Thus, the rat melatonin rMT(1) receptor was cloned using DNA fragments for exon 1 and 2 amplified from rat genomic DNA followed by screening of a rat genomic library for the full length exon sequences. The rat rMT(2) receptor subtype was cloned in a similar manner with the exception of exon 1 which was identified by screening a rat genomic library with exon 1 of the human hMT(2) receptor. The coding region of these receptors translates proteins of 353 and 364 amino acids, respectively, for rMT(1) and rMT(2). A 55% homology was observed between both rat isoforms. The entire contiguous rat MT1 and MT2 receptor coding sequences were cloned, stably expressed in CHO cells and characterized in binding assay using 2- [I-125]-Iodomelatonin. The dissociation constants (K-d) for rMT(1) and rMT(2) were 42 and 130 pM, respectively. Chemically diverse compounds previously characterized at human MT1 and MT2 receptors were evaluated at rMT1 and rMT2 receptors, for their binding affinity and functionality in [S-35] -GTP gamma S binding assay. Some, but not all, compounds shared a similar binding affinity and functionality at both rat and human corresponding subtypes. A different pharmacological profile of the MT1 subtype has also been observed previously between human and ovine species. These in vitro results obtained with the rat melatonin receptors are thus of importance to understand the physiological roles of each subtype in animal models. (C) 2008 Elsevier Inc. All rights reserved.

Original languageEnglish
Pages (from-to)2007-2019
Number of pages13
JournalBiochemical Pharmacology
Volume75
Issue number10
DOIs
Publication statusPublished - 15 May 2008

Keywords

  • melatonin
  • melatonin receptors
  • selective agonists
  • selective antagonists
  • rat
  • amidic derivatives
  • ligand binding
  • antagonists
  • agonists
  • design
  • identification
  • activation
  • affinity
  • retina
  • artery

Cite this

Audinot, V., Bonnaud, A., Grandcolas, L., Rodriguez, M., Nagel, N., Galizzi, J-P., ... Boutin, J. A. (2008). Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors. Biochemical Pharmacology, 75(10), 2007-2019. https://doi.org/10.1016/j.bcp.2008.02.022

Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors. / Audinot, Valerie; Bonnaud, Anne; Grandcolas, Line; Rodriguez, Marianne; Nagel, Nadine; Galizzi, Jean-Pierre; Balik, Ales; Messager, Sophie; Hazlerigg, David G.; Barrett, Perry; Delagrange, Philippe; Boutin, Jean A.

In: Biochemical Pharmacology, Vol. 75, No. 10, 15.05.2008, p. 2007-2019.

Research output: Contribution to journalArticle

Audinot, V, Bonnaud, A, Grandcolas, L, Rodriguez, M, Nagel, N, Galizzi, J-P, Balik, A, Messager, S, Hazlerigg, DG, Barrett, P, Delagrange, P & Boutin, JA 2008, 'Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors', Biochemical Pharmacology, vol. 75, no. 10, pp. 2007-2019. https://doi.org/10.1016/j.bcp.2008.02.022
Audinot V, Bonnaud A, Grandcolas L, Rodriguez M, Nagel N, Galizzi J-P et al. Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors. Biochemical Pharmacology. 2008 May 15;75(10):2007-2019. https://doi.org/10.1016/j.bcp.2008.02.022
Audinot, Valerie ; Bonnaud, Anne ; Grandcolas, Line ; Rodriguez, Marianne ; Nagel, Nadine ; Galizzi, Jean-Pierre ; Balik, Ales ; Messager, Sophie ; Hazlerigg, David G. ; Barrett, Perry ; Delagrange, Philippe ; Boutin, Jean A. / Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors. In: Biochemical Pharmacology. 2008 ; Vol. 75, No. 10. pp. 2007-2019.
@article{9742212169c942c29018dcd59a28433b,
title = "Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors",
abstract = "In order to interpret the effects of melatonin ligands in rats, we need to determine their activity at the receptor subtype level in the corresponding species. Thus, the rat melatonin rMT(1) receptor was cloned using DNA fragments for exon 1 and 2 amplified from rat genomic DNA followed by screening of a rat genomic library for the full length exon sequences. The rat rMT(2) receptor subtype was cloned in a similar manner with the exception of exon 1 which was identified by screening a rat genomic library with exon 1 of the human hMT(2) receptor. The coding region of these receptors translates proteins of 353 and 364 amino acids, respectively, for rMT(1) and rMT(2). A 55{\%} homology was observed between both rat isoforms. The entire contiguous rat MT1 and MT2 receptor coding sequences were cloned, stably expressed in CHO cells and characterized in binding assay using 2- [I-125]-Iodomelatonin. The dissociation constants (K-d) for rMT(1) and rMT(2) were 42 and 130 pM, respectively. Chemically diverse compounds previously characterized at human MT1 and MT2 receptors were evaluated at rMT1 and rMT2 receptors, for their binding affinity and functionality in [S-35] -GTP gamma S binding assay. Some, but not all, compounds shared a similar binding affinity and functionality at both rat and human corresponding subtypes. A different pharmacological profile of the MT1 subtype has also been observed previously between human and ovine species. These in vitro results obtained with the rat melatonin receptors are thus of importance to understand the physiological roles of each subtype in animal models. (C) 2008 Elsevier Inc. All rights reserved.",
keywords = "melatonin, melatonin receptors, selective agonists, selective antagonists, rat, amidic derivatives, ligand binding, antagonists, agonists, design, identification, activation, affinity, retina, artery",
author = "Valerie Audinot and Anne Bonnaud and Line Grandcolas and Marianne Rodriguez and Nadine Nagel and Jean-Pierre Galizzi and Ales Balik and Sophie Messager and Hazlerigg, {David G.} and Perry Barrett and Philippe Delagrange and Boutin, {Jean A.}",
year = "2008",
month = "5",
day = "15",
doi = "10.1016/j.bcp.2008.02.022",
language = "English",
volume = "75",
pages = "2007--2019",
journal = "Biochemical Pharmacology",
issn = "0006-2952",
publisher = "Elsevier Inc.",
number = "10",

}

TY - JOUR

T1 - Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors

AU - Audinot, Valerie

AU - Bonnaud, Anne

AU - Grandcolas, Line

AU - Rodriguez, Marianne

AU - Nagel, Nadine

AU - Galizzi, Jean-Pierre

AU - Balik, Ales

AU - Messager, Sophie

AU - Hazlerigg, David G.

AU - Barrett, Perry

AU - Delagrange, Philippe

AU - Boutin, Jean A.

PY - 2008/5/15

Y1 - 2008/5/15

N2 - In order to interpret the effects of melatonin ligands in rats, we need to determine their activity at the receptor subtype level in the corresponding species. Thus, the rat melatonin rMT(1) receptor was cloned using DNA fragments for exon 1 and 2 amplified from rat genomic DNA followed by screening of a rat genomic library for the full length exon sequences. The rat rMT(2) receptor subtype was cloned in a similar manner with the exception of exon 1 which was identified by screening a rat genomic library with exon 1 of the human hMT(2) receptor. The coding region of these receptors translates proteins of 353 and 364 amino acids, respectively, for rMT(1) and rMT(2). A 55% homology was observed between both rat isoforms. The entire contiguous rat MT1 and MT2 receptor coding sequences were cloned, stably expressed in CHO cells and characterized in binding assay using 2- [I-125]-Iodomelatonin. The dissociation constants (K-d) for rMT(1) and rMT(2) were 42 and 130 pM, respectively. Chemically diverse compounds previously characterized at human MT1 and MT2 receptors were evaluated at rMT1 and rMT2 receptors, for their binding affinity and functionality in [S-35] -GTP gamma S binding assay. Some, but not all, compounds shared a similar binding affinity and functionality at both rat and human corresponding subtypes. A different pharmacological profile of the MT1 subtype has also been observed previously between human and ovine species. These in vitro results obtained with the rat melatonin receptors are thus of importance to understand the physiological roles of each subtype in animal models. (C) 2008 Elsevier Inc. All rights reserved.

AB - In order to interpret the effects of melatonin ligands in rats, we need to determine their activity at the receptor subtype level in the corresponding species. Thus, the rat melatonin rMT(1) receptor was cloned using DNA fragments for exon 1 and 2 amplified from rat genomic DNA followed by screening of a rat genomic library for the full length exon sequences. The rat rMT(2) receptor subtype was cloned in a similar manner with the exception of exon 1 which was identified by screening a rat genomic library with exon 1 of the human hMT(2) receptor. The coding region of these receptors translates proteins of 353 and 364 amino acids, respectively, for rMT(1) and rMT(2). A 55% homology was observed between both rat isoforms. The entire contiguous rat MT1 and MT2 receptor coding sequences were cloned, stably expressed in CHO cells and characterized in binding assay using 2- [I-125]-Iodomelatonin. The dissociation constants (K-d) for rMT(1) and rMT(2) were 42 and 130 pM, respectively. Chemically diverse compounds previously characterized at human MT1 and MT2 receptors were evaluated at rMT1 and rMT2 receptors, for their binding affinity and functionality in [S-35] -GTP gamma S binding assay. Some, but not all, compounds shared a similar binding affinity and functionality at both rat and human corresponding subtypes. A different pharmacological profile of the MT1 subtype has also been observed previously between human and ovine species. These in vitro results obtained with the rat melatonin receptors are thus of importance to understand the physiological roles of each subtype in animal models. (C) 2008 Elsevier Inc. All rights reserved.

KW - melatonin

KW - melatonin receptors

KW - selective agonists

KW - selective antagonists

KW - rat

KW - amidic derivatives

KW - ligand binding

KW - antagonists

KW - agonists

KW - design

KW - identification

KW - activation

KW - affinity

KW - retina

KW - artery

U2 - 10.1016/j.bcp.2008.02.022

DO - 10.1016/j.bcp.2008.02.022

M3 - Article

VL - 75

SP - 2007

EP - 2019

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

IS - 10

ER -