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C Pilon

Publications and source records attributed to C Pilon.

At least 37 records · Page 2Linked to original sources
PubMed1996

Antipsychotics with inverse agonist activity at the dopamine D3 receptor.

In NG 108-15 cells expressing the recombinant human D3 receptor, dopamine agonists enhance [3H]thymidine incorporation and decrease cAMP accumulation. In these cells, but not in wild type cells, haloperidol, fluphenazine, and various other antipsychotics inhibited basal [3H]thymidine incorporation in a concentration-dependent manner. In contrast, other dopamine antagonists such as nafadotride or (+)AJ 76, two D3-preferring antagonists, were without effect. The concentration-response curve of haloperidol was shifted to the right in presence of nafadotride, with a potency compatible with its nanomolar apparent affinity as neutral antagonist. Pertussis toxin treatment abolished or markedly reduced the responses to haloperidol or fluphenazine. In contrast, no significant enhancement of cAMP accumulation could be observed, under the influence of haloperidol or eticlopride. These data indicate that some dopamine antagonists behave as inverse agonists, and thus appear to inhibit an agonist-independent activity of the D3 receptor on [3H]thymidine incorporation pathway, but not on the cAMP pathway.

Animals↗
PubMed1996

Effects of taxol on the human NCI-H295 adrenocortical carcinoma cell line.

We investigated the effects of taxol, an antimicrotubule agent active in different cancers, on the human NCI-H295 steroid-secreting adrenocortical carcinoma cell line. Cells were incubated for 48, 72 or 96 h with taxol 10(-10)-10(-4) M. Cell viability was evaluated by MTT assay with IC50 calculation. Apoptosis was investigated by measuring DNA fragmentation with ELISA assay after cell exposure to taxol at IC50 for 24 h. For secretion studies, aldosterone, cortisol, testosterone and dehydroepiandrosterone-sulphate (DHEA-S) were measured by RIA in the conditioned medium after 96 h exposure to taxol 10(-10)-10(-6) M, and expressed as percentage of steroid production by control cells. By MTT, taxol induced a dose-dependent inhibition of cell proliferation, with ICs50 at 72-96 h corresponding to blood levels achieved in vivo in patients with other types of cancer. Nuclear fragmentation, morphologically confirmed at electron microscopy, showed a 4-fold increase after exposure to taxol. With 10(-6) M taxol, aldosterone decreased to 48%, cortisol to 61%, testosterone to 76% and DHEA-S to 89% of steroid production by control cells. Taxol is an effective cytotoxic and antiproliferative agent in a human adrenocortical steroid-secreting carcinoma cell line. Apoptosis induced by the drug is involved in neoplastic cell death.

Adrenal Cortex Neoplasms↗
PubMed1995

The preferential dopamine D3 receptor ligand, (+)-UH232, is a partial agonist.

In a NG 108 15 hybrid cell line stably expressing a recombinant dopamine D3 receptor, (+)-UH 232 (cis-(+)-1S,2R)-5-methoxy-1-methyl-2-(di-n- propylamino)tetralin), a partially selective D3 receptor ligand, stimulates mitogenesis, as measured by incorporation of [3H]thymidine, with an EC50 of 7.6 nM and a maximal increase corresponding to 23% of the response elicited by quinpirole, a full agonist. This effect was antagonised by nafadotride, a D3 receptor-selective antagonist. (+)-UH 232 also antagonised quinpirole-induced mitogenesis with a Ki value of 9.4 nM. (+)-UH 232 (1 microM) inhibited by 22% the forskolin-induced accumulation of cAMP, whilst the inhibition by quinpirole (100 nM) was 53%. These results indicate that (+)-UH 232 is a partial agonist at the D3 receptor with an intrinsic activity of 0.2-0.4.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1995

A functional test identifies dopamine agonists selective for D3 versus D2 receptors.

The functional potency of a series of dopamine agonists for increasing mitogenesis, measured by incorporation of [3H]thymidine, was established in transfected cell lines expressing human D2 or D3 receptors. The functional selectivity of agonists markedly differs from their binding selectivity. (+)7-OH-DPAT, pramipexole, quinerolane and PD 128,907, the most D3 receptor-selective compounds in binding studies, were 7, 15, 21 and 54 times more potent, respectively, at the D3 than at the D2 receptor in the functional test. Bromocriptine displayed a 10-fold functional selectivity toward the D2 receptor. The known behavioural actions of D3 selective agonists support a role for the D3 receptor in motor inhibitions, which should be taken into account for the treatment of motor dysfunctions by dopamine agonists.

Animals↗
PubMed1995

Experience of immediate stenting, perfusion balloon and secondary stenting in failed angioplasty.

OBJECTIVE: To present a comparative analysis of the immediate and short term experience of stent implantation as the initial treatment for failed angioplasty (immediate stenting) with the staged approach of prolonged inflation with a perfusion balloon followed by stent implantation if prolonged inflation was unsuccessful (secondary stenting). DESIGN: Retrospective analysis of the outcome of 90 consecutive patients from a single institution undergoing one of these treatment strategies for failed angioplasty. INTERVENTIONS AND MAIN RESULTS: The initial treatment for failed angioplasty was prolonged inflation with a perfusion balloon in 59 and immediate stent implantation in 31 patients. Angiographic success (less than 50% residual and normal flow) was achieved in 24 of 59 (41%) with the perfusion balloon (mean duration of inflation 12.2 +/- 8.3 mins) and in 30 of 31 (97%) with immediate stent implantation (P = 0.0001). Of the 35 patients in whom the perfusion balloon was unsuccessful secondary stenting was attempted in 27, with angiographic success in 24 (89%). Stent thrombosis occurred in four (13%) of the immediate stent group and in three (13%) of the secondary stent group. Acute ischemia compatible with vessel occlusion occurred in one (4%) of the perfusion balloon alone group following an initially successful result. One month following the procedure, 23 (72%) of the patients treated with perfusion balloon alone, 26 (84%) of the immediate stent group and 21 (78%) of the secondary stent group had a successful outcome and were free from major complications. CONCLUSION: Immediate stent implantation and the staged approach of prolonged inflation with a perfusion balloon followed by secondary stent implantation if prolonged inflation was unsuccessful yielded similar results.

Angioplasty, Balloon, Coronary↗
PubMed1994

Functional coupling of the human dopamine D3 receptor in a transfected NG 108-15 neuroblastoma-glioma hybrid cell line.

Transfection of a human dopamine D3 receptor cDNA in a neuroblastoma-glioma hybrid cell line (NG 108-15) provided clonal cell lines stably expressing up to 600 fmol per mg protein of [125I]iodosulpiride binding sites. Dopamine and several agonists distinguished two receptor-affinity states in membranes. In the case of dopamine, the high-affinity state (Ki = 0.9 nM, 30% of total binding) was completely converted into a low-affinity state (Ki = 57 nM) in the presence of 10 microM guanosine-5'-O-(3-thiotriphosphate). In addition to these two sites, a site with a very low affinity for dopamine was evidenced in whole cells. The dopamine D3 receptor mediated two responses: c-fos activation, as measured by the appearance of Fos-like immunoreactivity, and increased mitogenesis, as measured by incorporation of [3H]thymidine. The Fos-like immunoreactivity appeared within 30 min, lasted 2 h and was blocked by the partially selective dopamine D3 receptor compound (+)-UH 232 (cis-(+)-5-methoxy-1-methyl-2-(di-n-propylamino)tetralin). The mitogenic effect, which occurred after a lag time (over 2 h stimulation), was produced with subnanomolar potency and full intrinsic activity by several compounds previously identified as dopamine D2 receptor agonists, e.g. quinpirole, (+)-7-OH-DPAT ((+)-7-hydroxy-2-(di-n-propylamino)tetralin) and RU 24926 (N-n-propyl-di-beta(3-hydroxyphenyl)-ethylamine), and was reversibly blocked by (+)-UH 232 (Ki = 9 nM). Talipexole (B-HT 920, 5-allyl-2-amino-5,6,7,8-tetrahydro-4H-thiazolo[4,5-d]azepin) was identified as a partial agonist at the dopamine D3 receptor. Dopamine D3 receptor-mediated mitogenesis was potentiated by a phorbol ester and was abolished by pretreatment with pertussis toxin. A mitogenic effect of same amplitude was elicited by bradykinin or carbachol, both acting through constitutive receptors. Bradykinin markedly activated inositol phosphate turnover, and had no effect on forskolin-stimulated cyclic AMP accumulation. Carbachol inhibited forskolin-stimulated cyclic AMP accumulation and had no effect on inositol-phosphate turnover. Quinpirole had no effect on any of these second messenger pathways. Thus, in transfected NG 108-15 cells, the dopamine D3 receptor is coupled to a pertussis toxin-sensitive G protein and mediates two possibly unrelated biological effects, through initial biochemical events that remain to be identified.

Animals↗
PubMed1992

Identification, characterization, and localization of the dopamine D3 receptor in rat brain using 7-[3H]hydroxy-N,N-di-n-propyl-2-aminotetralin.

We have identified 7-[3H]hydroxy-N,N-di-n-propyl-2-aminotetralin ([3H]7-OH-DPAT) as a selective probe for the recently cloned dopamine D3 receptor and used it to assess the presence of this receptor and establish its distribution and properties in brain. In transfected Chinese hamster ovary (CHO) cells, it binds to D3 receptors with subnanomolar affinity, whereas its affinity is approximately 100-, 1000-, and 10,000-fold lower at D2, D4, and D1 receptors, respectively. Specific [3H]7-OH-DPAT binding sites, with a Kd of 0.8 nM and a pharmacology similar to those at reference D3 receptors of CHO cells, were identified in rat brain. D3 receptors differ from D2 receptors in brain by their lower abundance (2 orders of magnitude) and distribution, restricted to a few mainly phylogenetically ancient areas--e.g., paleostriatum and archicerebellum--as evidenced by membrane binding are autoradiography studies. Native D3 receptors in brain are characterized by an unusually high nanomolar affinity for dopamine and a low modulatory influence of guanyl nucleotides on agonist binding. These various features suggest that D3 receptors are involved in a peculiar mode of neurotransmission in a restricted subpopulation of dopamine neurons.

Animals↗
PubMed1992

Pharmacology of human dopamine D3 receptor expressed in a mammalian cell line: comparison with D2 receptor.

Two cell lines were created by transfecting cDNAs of the human D2 receptor or the recently cloned human D3 receptor to CHO cells, and the properties of [125I]iodosulpride binding to membranes of these cells were compared. In cell lines expressing the D2 receptor subtype where the selectable marker, a phleomycin-resistance gene, was cotransfected in a different plasmid, a stable expression could be maintained for only few passages. In cell lines expressing the D3 receptor subtype, the selectable marker, a dihydrofolate reductase gene, was cotransfected in the same plasmid and a stable expression could be obtained. In addition, the D3 receptor gene could be amplified in these latter cell lines and a high expression level reached (up to 10(6) binding sites per cell). Sodium and, to a lesser extent, lithium similarly increased [125I]iodosulpride binding to D2 and D3 receptors. In the absence of guanylnucleotide, dopamine had a 24-fold higher apparent affinity at D3 than at D2 receptors. Gpp(NH)p induced rightward shift and steepening of dopamine competition curves at either subtype but the effects were more marked at D2 than at D3 receptors. Several agonists and antagonists, previously regarded as autoreceptor-selective, displayed higher affinities at D3 than at D2 receptors. Although most antagonists used as antipsychotics displayed high affinities at the D3 receptor, all were more potent at the D2 receptor. However, the ratio of Ki values varied over about 10-fold among these compounds, suggesting that they realize differential dopamine receptor subtype occupancy during treatments and that this might be reflected in their clinical profile.

Animals↗
PubMed1992

Localization and function of the D3 dopamine receptor.

A novel dopamine receptor has been recently cloned, which differs from D1 and D2 receptors by its sequence, localization, pharmacology and possibly signalling system, hence its designation as D3 receptor. The D3 receptor cDNA was obtained by a combination of screenings of DNA libraries and PCR polymerase chain reaction experiments. It encodes a protein with a predicted structure consisting in 7 transmembrane domains indicating that it belongs to the G-protein coupled receptor family. Its global homology is 52% with the D2 receptor but 78% if only transmembrane domains are considered. Two shorter transcript variants, in addition to the full-length cDNA were detected by PCR in various rat brain regions. The shorter transcripts are generated by alternative splicing and encode two putative proteins respectively interrupted after the second transmembrane domain and lacking the second extracytoplasmic loop. After transfection of this latter isoform into cultured cells, no dopaminergic activity could be detected. These shorter splice variants may regulate the number of active D3 receptors. The human D3 receptor was also cloned using similar approaches with rat D3 receptor cDNA probes and was found highly homologous to the rat receptor, except in the third intracytoplasmic loop. The human D3 receptor gene was assigned to the chromosome 3 at q13.3 band. Visualisation of D3 receptor mRNA in rat brain by in situ hybridization indicated a predominant expression of the message in the ventral striatum and other "limbic" areas. There is no overlap in the distributions of D2 and D3 receptor mRNAs in discrete structures, suggesting that the two transcripts are expressed by different cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1991

Dopamine activation of the arachidonic acid cascade as a basis for D1/D2 receptor synergism.

Understanding the actions of the neurotransmitter dopamine in the brain is important in view of its roles in neuropsychiatric illnesses. Dopamine D1 receptors, which stimulate both adenylyl cyclase and phospholipase C, and D2 receptors, which inhibit them, can nevertheless act synergistically to produce many electrophysiological and behavioral responses. Because this functional synergism can occur at the level of single neurons, another, as yet unidentified, signalling pathway activated by dopamine has been hypothesized. We report here that in Chinese hamster ovary (CHO) cells transfected with the D2 receptor complementary DNA, D2 agonists potently enhanced arachidonic acid release, provided that such release has been initiated by stimulating constitutive purinergic receptors or by increasing intracellular Ca2+. In CHO cells expressed D1 receptors, D1 agonists exert no such effect. When D1 and D2 receptors are coexpressed, however, activation of both subtypes results in a marked synergistic potentiation of arachidonic acid release. The numerous actions of arachidonic acid and its metabolites in neuronal signal transduction suggest that facilitation of its release may be implicated in dopaminergic responses, such as feedback inhibition mediated by D2 autoreceptors, and may constitute a molecular basis for D1/D2 receptor synergism.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1991

Shorter variants of the D3 dopamine receptor produced through various patterns of alternative splicing.

Using Polymerase Chain Reaction amplification of mRNAs from several areas of rat brain we have shown the occurrence of two shorter transcripts of the dopamine D3 receptor gene, in addition to that corresponding to the D3 receptor. Cloning and sequencing of these transcripts, together with the establishment of the exon-intron organization of the D3 receptor gene, shown these transcripts to result from different processes of alternative splicing. The first transcript encodes a 100 amino acid protein, being produced by splicing of an exon whose absence deletes the third transmembrane domain and gives rise downstream to a frameshift in the open reading frame. In the second transcript, an in frame 54 bp deletion is produced by splicing occurring at an internal acceptor site, suppressing half of the second extracellular loop and a small sequence in the fifth transmembrane domain. This transcript was stably expressed in CHO cells which, however, failed to reveal any dopaminergic ligand binding activity. The functional significance and possible role of these shorter variants of the dopamine D3 receptor in cell signalling remain to be established.

Amino Acid Sequence↗
PubMed1991

Chromosomal localization of the human D3 dopamine receptor gene.

A novel dopamine D3 receptor gene that may be involved in psychiatric diseases has recently been characterized. It has been assigned to chromosome 3 by hybridization with a D3 receptor probe to human sorted chromosomes, and localized to band 3q 13.3 by in situ hybridization.

Chromosome Mapping↗
PubMed1989

Organization of the human lipoprotein lipase gene and evolution of the lipase gene family.

The human lipoprotein lipase gene was cloned and characterized. It is composed of 10 exons spanning approximately equal to 30 kilobases. The first exon encodes the 5'-untranslated region, the signal peptide plus the first two amino acids of the mature protein. The next eight exons encode the remaining 446 amino acids, and the tenth exon encodes the long 3'-untranslated region of 1948 nucleotides. The lipoprotein lipase transcription start site and the sequence of the 5'-flanking region were also determined. We compared the organization of genes for lipoprotein lipase, hepatic lipase, pancreatic lipase, and Drosophila yolk protein 1, which are members of a family of related genes. A model for the evolution of the lipase gene family is presented that involves multiple rounds of gene duplication plus exon-shuffling and intron-loss events.

Amino Acid Sequence↗