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Research ArticleArticle

A Na+ /Cl−-Dependent Transporter for Catecholamines, Identified as a Norepinephrine Transporter, Is Expressed in the Brain of the Teleost Fish Medaka (Oryzias latipes)

Christine Roubert, Corinne Sagné, Marika Kapsimali, Philippe Vernier, Frank Bourrat and Bruno Giros
Molecular Pharmacology September 2001, 60 (3) 462-473;
Christine Roubert
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Corinne Sagné
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Marika Kapsimali
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Philippe Vernier
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Frank Bourrat
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Bruno Giros
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Abstract

We report the isolation, functional characterization, and localization of a Na+/Cl−-dependent catecholamine transporter (meNET) present in the brain of the teleost fish medaka. This carrier is very similar to the human neuronal norepinephrine transporter (NET) and the human neuronal dopamine transporter (DAT), showing 70 and 64% amino acid identity, respectively. When expressed in COS-7 cells, this transporter mediates the high-affinity uptake of dopamine (KM = 290 nM) and norepinephrine (KM = 640 nM). Its pharmacological profile reveals more similarities with NET, including a high affinity for the tricyclic antidepressants desipramine (IC50 = 0.92 nM) and nortriptyline (IC50 = 16 nM). In situ hybridization on the medaka brain shows that meNET mRNA is present only in a subset of tyrosine hydroxylase-positive neurons found in the noradrenergic areas of the hindbrain, such as the locus ceruleus and area postrema. None of the dopaminergic areas anterior to the isthmus contains any labeled neurons. Neither reverse transcriptase-polymerase chain reaction with degenerate primers specific for γ-aminobutyric acid transporter/NET nor autoradiographic experiments with [125I]3b-(4-iodophenyl)-tropane-2b-carboxylic acid methyl ester revealed an additional catecholamine transporter in the medaka brain. Uptake experiments with medaka brain synaptosomes show an endogenous transport with a pharmacological profile identical to that of the recombinant meNET. Thus, meNET is probably the predominant—if not the only—catecholamine transporter in the medaka fish brain. In view of the highly conserved primary structures and pharmacological properties of meNET, it is tempting to speculate that a specific dopamine transport developed later in vertebrate evolution and probably accompanied the tremendous enlargement of the meso-telencephalic dopaminergic pathways in amniotes.

Footnotes

  • ↵1 Present address: Sanofi-Synthelabo, Département SNC, 34184 Montpellier Cedex 04, France.

  • This research was supported by grants from Institut National de la Santé et de la Recherche Médicale (to B.G.) and from Centre National de la Recherche Scientifique and University Paris XI (to P.V.). C.R. was supported by Sanofi Recherche and Fondation pour la Recherche Medicale. M.K. was supported by Lilly and Fondation pour la Recherche Medicale. C.S. was supported by Rhône-Poulenc Rorer and Association France Parkinson.

  • Abbreviations:
    DA
    dopamine
    NE
    norepinephrine
    DAT
    dopamine transporter
    NET
    norepinephrine transporter
    TMD
    transmembrane domain(s)
    fET
    Rana catesbiana peripheral epinephrine transporter
    ceDAT
    Caenorhabditis elegans dopamine transporter
    RTI-55
    3b-(4-iodophenyl)-tropane-2b-carboxylic acid methyl ester
    meNET
    medaka norepinephrine transporter
    PCR
    polymerase chain reaction
    SSC
    standard saline citrate
    RT
    reverse transcriptase
    PBS
    phosphate-buffered saline
    SERT
    serotonin transporter
    hNET
    human norepinephrine transporter
    GAT
    γ-aminobutyric acid transporter
    GABA
    γ-aminobutyric acid
    rDAT
    rat dopamine transporter
    TH
    tyrosine hydroxylase
    PKC
    protein kinase C
    • Received March 2, 2001.
    • Accepted May 25, 2001.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 60 (3)
Molecular Pharmacology
Vol. 60, Issue 3
1 Sep 2001
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Research ArticleArticle

A Na+ /Cl−-Dependent Transporter for Catecholamines, Identified as a Norepinephrine Transporter, Is Expressed in the Brain of the Teleost Fish Medaka (Oryzias latipes)

Christine Roubert, Corinne Sagné, Marika Kapsimali, Philippe Vernier, Frank Bourrat and Bruno Giros
Molecular Pharmacology September 1, 2001, 60 (3) 462-473;

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Research ArticleArticle

A Na+ /Cl−-Dependent Transporter for Catecholamines, Identified as a Norepinephrine Transporter, Is Expressed in the Brain of the Teleost Fish Medaka (Oryzias latipes)

Christine Roubert, Corinne Sagné, Marika Kapsimali, Philippe Vernier, Frank Bourrat and Bruno Giros
Molecular Pharmacology September 1, 2001, 60 (3) 462-473;
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