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Titolo:
Identification of dopamine plasma membrane and vesicular transporters in human peripheral blood lymphocytes
Autore:
Amenta, F; Bronzetti, E; Cantalamessa, F; El-Assouad, D; Felici, L; Ricci, A; Tayebati, SK;
Indirizzi:
Univ Camerino, Dipartimento Sci Farmacol & Med Sperimentale, Sez Anat Umana, I-62032 Camerino, Italy Univ Camerino Camerino Italy I-62032 Anat Umana, I-62032 Camerino, Italy Univ Roma La Sapienza, Dipartimento Sci Cardiovasc & Resp, I-00161 Rome, Italy Univ Roma La Sapienza Rome Italy I-00161 asc & Resp, I-00161 Rome, Italy
Titolo Testata:
JOURNAL OF NEUROIMMUNOLOGY
fascicolo: 1-2, volume: 117, anno: 2001,
pagine: 133 - 142
SICI:
0165-5728(20010702)117:1-2<133:IODPMA>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
RADIOLIGAND BINDING ASSAY; PARKINSONS-DISEASE; RECEPTOR; EXPRESSION; SYSTEM;
Keywords:
dopamine transporter; vesicular monoamine transporter; lymphocytes; radioligand binding assay; Western blot analysis; immunocytochemistry;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
33
Recensione:
Indirizzi per estratti:
Indirizzo: Amenta, F Univ Camerino, Dipartimento Sci Farmacol & Med Sperimentale, SezAnat Umana, Via Scalzino 3, I-62032 Camerino, Italy Univ Camerino Via Scalzino 3 Camerino Italy I-62032 rino, Italy
Citazione:
F. Amenta et al., "Identification of dopamine plasma membrane and vesicular transporters in human peripheral blood lymphocytes", J NEUROIMM, 117(1-2), 2001, pp. 133-142

Abstract

Plasma membrane dopamine transporter (DAT), vesicular monoamine transporters (VMAT) type-1 and -2 and the expression of the dopaminergic markers dopamine and tyrosine hydroxylase were assessed in membranes and/or in cytospincentrifuged human peripheral blood lymphocytes. The radiolabeled DAT ligand [H-3]GBR12935 was bound to peripheral lymphocytes in a manner consistent with the specific binding to a dopamine uptake system, with a dissociation constant similar to that found in striatum, but with a lower density of binding sites. On the other hand, no specific binding occurred in cerebellum used as a test tissue not expressing DAT. Western blot analysis using antibodies raised against amino or carboxy terminus of DAT or against VMAT-1 or VMAT-2 revealed labeling of single bands of approximately 76, 55 or 68 KDa, respectively, displaying similar migration characteristics in lymphocytes and test tissues used for comparison. Immunofluorescence revealed that anti-dopamine, anti-tyrosine hydroxylase, anti-DAT, anti-VMAT-1 and anti-VMAT-2 antibodies labeled the total population of cytospin-centrifuged lymphocytesmounted on microscope slides. Confocal laser microscopy demonstrated that dopamine and VMAT-2 immunoreactivity was developed mainly in cytoplasmic punctiform areas likely corresponding to vesicles and to a lower extent was associated to plasma membrane. Tyrosine hydroxylase immunoreactivity was diffused to cytoplasm and to plasma membrane of lymphocytes, whereas DAT and VMAT-1 immunoreactivity were located almost exclusively in lymphocyte plasmamembrane and cytoplasm, respectively. Lymphocyte DAT characterized in thisstudy has probably functional relevance as [H-3]dopamine was taken up by intact lymphocytes and uptake was inhibited specifically by compounds known to affect dopamine transport. These findings indicate that human peripheralblood lymphocytes possess DAT plasma membrane and VMAT-1 and VMAT-2 transporters. Increasing evidence indicates that dopamine transporter changes maybe related to neuronal injury. In view of this assessment of lymphocyte DAT and VMAT transporters can be considered for identifying pathologies characterized by impaired dopaminergic neurotransmission. (C) 2001 Elsevier Science B.V. All rights reserved.

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Documento generato il 04/04/20 alle ore 07:41:06