Characterization of neurotensin binding sites on rat mesencephalic cells in primary culture

Dana, C; Pélaprat, Didier; Vial, Micheline; Brouard, Aline; Lhiaubet, Anne-Marie; Rostène, William

doi:10.1016/0165-3806(91)90139-a

Cited by 14 publications

(13 citation statements)

References 41 publications

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“…Furthermore, high-affinity NT receptors in this area have been shown to be selectively associated with dopaminergic (DA) neurons in both rodent and primate brain (Palacios and Kuhar, 1982;Sadoul et al, 1984;Quirion et al, 1987;Szigethy and Beaudet, 1989). In keeping with these results, rat embryonic DA neurons were found to be enriched with functional NT receptors in primary cell cultures from the rat midbrain (Chabry et al, 1990;Dana et al, 1991;Brouard et al, 1992). Pure glial cultures were taken from the neocortex of neonatal animals, because this region was reported to express among the highest levels of low-affinity NT receptors in the adult rat brain (Chalon et al, 1996;Mazella et al, 1996).…”

Section: Abstract: Confocal Microscopy; Immunohistochemistry; Tyrosisupporting

confidence: 69%

Differential Binding Profile and Internalization Process of Neurotensin via Neuronal and Glial Receptors

et al. 1997

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Two G-protein-coupled receptors for the tridecapeptide neurotensin (NT) have been identified and cloned in mammalian brain: a high-affinity (K d ϭ 0.3 nM) receptor, sensitive to the antagonist SR 48692 but insensitive to levocabastine, and a lower-affinity (K d ϭ 2-4 nM) receptor, sensitive to levocabastine but with poor affinity for SR 48692. Although there is good evidence that the high-affinity site is predominantly expressed in neurons, little is known of the cellular localization of the low-affinity receptor. In the present study, we identify by confocal microscopy selective levocabastine-sensitive, SR 48692-resistant binding of a fluorescent derivative of NT (fluo-NT) to a subpopulation of glial fibrillary acidic protein-immunoreactive glial cells grown in culture from the midbrain and cerebral cortex of embryonic and neonatal rats, respectively. We also demonstrate, by combining fluo-NT detection with tyrosine hydroxylase immunofluorescence, that these glial binding sites are differentially regulated from the SR 48692-sensitive NT receptor expressed in the same cultures by mesencephalic dopamine neurons. Whereas the latter undergoes rapid ligandinduced internalization followed by centripetal mobilization of ligand-receptor complexes from processes to perikarya and from perikaryal periphery to cell center, the former induces the formation of cell-surface clusters that fail to internalize. It is concluded that NT may exert its effects on both neurons and astrocytes in the CNS. Whereas NT neural signaling is exerted through high-affinity receptors and may be partly effected through internalization of receptor-ligand complexes, glial signaling is exerted through low-affinity NT receptors and appears to be transduced exclusively at the level of the plasma membrane.

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Section: Abstract: Confocal Microscopy; Immunohistochemistry; Tyrosisupporting

confidence: 69%

Differential Binding Profile and Internalization Process of Neurotensin via Neuronal and Glial Receptors

et al. 1997

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“…The tion, autoradiograms are almost indiscernible from the film background (Fig. 2) (20), and COS-7 cells transfected with the cloned high-affinity rat brain receptor (18). SR 48692 also potently inhibited l251-neurotensin binding to the high-affinity binding sites in adult rat and mouse brain homogenates, whereas it was much less potent at the low-affinity, levocabastine-sensitive binding sites that are also present in these preparations (16).…”

Section: Pre~ara~on Of Timm S Whole Brain Homoge-mentioning

confidence: 99%

“…All binding assays with tissue homogenates were carried out at 209C in 50 mM Tris HCl buffer (pH 7.5) containing 0.2% bovine serum albumin, and 1 mM 1,10-phenanthroline (newborn mouse and human brain, technique as described (19,21). Binding experiments with rat mesencephalic neurons (500,000 cells per well) were carried out at 37TC as reported (20 maximal neurotensin-induced increase in the fluorescence ratio was determined.…”

Section: Pre~ara~on Of Timm S Whole Brain Homoge-mentioning

confidence: 99%

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Biochemical and pharmacological profile of a potent and selective nonpeptide antagonist of the neurotensin receptor.

Gully

Canton²,

Boigegrain³

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

373

251

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We describe the characteristics of SR 48692, a selective, nonpeptide antagonist of the neurotensin receptor. In vitro, this compound competitively inhibits 125I-labeled devoid of any intrinsic agonist activity. This compound is also active in vivo, since it reverses at low dose (80 pg/kg) the turning behavior induced by intrastriatal injection of neurotensin in mice with similar potency whatever the route of administration (i.p. or orally) and with a long duration of action (6 hr). Thus, being a potent and selective neurotensin receptor antagonist, SR 48692 may be considered as a powerful tool for investigating the role of neurotensin in physiological and pathological processes.

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“…Neurotensin (NT) and its receptors are present in high density in the ventral mesencephalon (Palacios and Kuhar, 1981;Szigethy and Beaudet, 1989;Dana et al, 1991). They are also present on astrocytes (Nouel et al, 1999;Trudeau, 2000).…”

Section: Introductionmentioning

confidence: 99%

Role of Calcium in Neurotensin-Evoked Enhancement in Firing in Mesencephalic Dopamine Neurons

St-Gelais¹,

Legault²,

Bourque³

et al. 2004

J. Neurosci.

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Characterization of neurotensin binding sites on rat mesencephalic cells in primary culture

Cited by 14 publications

References 41 publications

Differential Binding Profile and Internalization Process of Neurotensin via Neuronal and Glial Receptors

Differential Binding Profile and Internalization Process of Neurotensin via Neuronal and Glial Receptors

Biochemical and pharmacological profile of a potent and selective nonpeptide antagonist of the neurotensin receptor.

Role of Calcium in Neurotensin-Evoked Enhancement in Firing in Mesencephalic Dopamine Neurons

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