Elsevier

Brain Research

Volume 679, Issue 1, 8 May 1995, Pages 82-88
Brain Research

Cloning and characterization of the promoter region of the mouse μ opioid receptor gene

https://doi.org/10.1016/0006-8993(95)00222-CGet rights and content

Abstract

Opioid compounds have potent analgesic and euphoric properties. They act with specific cell-membrane receptors which have been pharmacologically defined into three major classes, μ, κ and δ. These receptors are highly regulated with respect to their gene expression, resulting in a temporally and spatially specific pattern of distribution for each receptor. To characterize the promoter sequence of the μ opioid receptor (MOR) gene, a mouse genomic DNA library was screened under high stringency with a rat MOR (MOR-1) cDNA probe and genomic sequences for the mouse MOR gene were isolated. From one genomic clone, a 2.3-kbEcoRI fragment, which hybridized to the 5′-end of the rat MOR-1 cDNA probe, was subcloned and sequenced. This fragment contains 1.3 kb of sequence upstream of the initiation codon, extends downstream through exon 1 and includes a portion of intron 1. Primer extension analysis using mouse brain poly (A)+ RNA identified a transcription initiation site 793 bp upstream from the translation start site. Chimeric constructs of mouse MOR deletion fragments fused to a luciferase reporter gene were transfected into a human neuroblastoma cell line, SK-N-SH, which constitutively expresses endogenous MOR. These transient expression studies indicated that the 0.2-kb region upstream from the transcription site possesses a functional promoter, which directs the expression of the reporter gene in vitro and may possess promoter activity for the mouse MOR gene in vivo.

References (35)

  • ThompsonR.C. et al.

    Cloning and pharmacological characterization of a rat μ opioid receptor

    Neuron

    (1993)
  • WangJ.-B. et al.

    Human μ opiate receptor cDNA and genomic clones, pharmacologic characterization and chromosomal assignment

    FEBS Lett.

    (1994)
  • YuV.C. et al.

    Human neuroblastoma cell line expresses μ and δ opioid receptor sites

    J. Biol. Chem.

    (1986)
  • ChenY. et al.

    Molecular cloning and functional expression of a μ-opioid receptor from rat brain

    Mol. Pharmacol.

    (1993)
  • ChildersS.R.

    Opioid receptor-coupled second messenger systems

    Handb. Exp. Pharmacol.

    (1993)
  • CorbettA.D. et al.

    Selectivity of ligands for opioid receptors

    Handb. Exp. Pharmacol.

    (1993)
  • CoxB.M.

    Opioid receptor-G protein interactions: acute and chronic effects of opioids

    Handb. Exp. Pharmacol.

    (1993)
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