Biomedical subjects

J Vignon

Publications and source records attributed to J Vignon.

At least 19 recordsLinked to original sources

PubMed2004

P2Y purinergic potentiation of glucose-induced insulin secretion and pancreatic beta-cell metabolism.

Purine nucleotides and their analogs increase insulin secretion through activation of pancreatic beta-cell P2Y receptors. The present study aimed at determining the role of glucose metabolism in the response to P2Y agonists and whether ATP-activated K+ channels (KATP channels) are involved in this response. The experiments were performed in the rat isolated pancreas, perfused with a Krebs-bicarbonate buffer supplemented with 2 g/l bovine serum albumin under dynamic glucose conditions from 5 mmol/l baseline to 11 mmol/l. ADPbetaS (0.5 micromol/l) was selected as a stable and selective P2Y agonist. This compound, ineffective on the 5 mmol/l glucose background, induced a significant threefold increase in insulin release triggered by the glucose challenge. The effect of ADPbetaS was markedly reduced (P <0.001) in the presence of an inhibitor of glucose metabolism. In addition to glucose, the ADP analog also amplified the beta-cell insulin response to 15 mmol/l methyl pyruvate (P <0.05), but it was ineffective on the insulin response to 2.5 mmol/l methyl succinate. A nonmetabolic stimulus was applied using tolbutamide (185 micromol/l). Insulin secretion induced by the KATP channel blocker was strongly reinforced by ADPbetaS (P <0.001), which prompted us to check a possible interplay of KATP channels in the effect of ADPbetaS. In the presence of diazoxide 250 micromol/l and 21 mmol/l KCl, ADPbetaS still amplified the second phase of glucose-induced insulin secretion (P <0.001). We conclude that P2Y receptor activation is able to promote insulin secretion through a mechanism, involving beta-cell metabolism and a rise in intracellular calcium; this effect does not result from a direct inhibitory effect on KATP channels.

Adenosine Diphosphate↗

PubMed2001

Interaction of gacyclidine enantiomers with 'non-NMDA' binding sites in the rat central nervous system.

Gacyclidine, a channel blocker of N-methyl-D-aspartate receptors (NMDAR), exhibits potent neuroprotective properties and a low self-neurotoxicity. Preventing its interaction with NMDARs we demonstrate, through the use of its enantiomers, that gacyclidine also interacts with other ('non-NMDA') binding sites. The autoradiographic study showed that these sites displayed a uniform specific binding in the forebrain and a more discrete distribution in the molecular layer of the cerebellum. The 'non-NMDA' binding sites could exert a modulatory control on glutamatergic neurotransmission.

Animals↗

PubMed2001

Characterization of 'non-N-methyl-D-Aspartate' binding sites for gacyclidine enantiomers in the rat cerebellar and telencephalic structures.

Gacyclidine is a non-competitive NMDA receptor antagonist with potent neuroprotective properties. However, we have previously demonstrated that gacyclidine enantiomers [(-) and (+)GK11] interact with other ('non-NMDA') binding sites which may play a role in the lower self-neurotoxicity of this compound relative to the other NMDA receptor antagonists. Evidence for these binding sites has been obtained from autoradiographic and membrane binding experiments. They were found to be expressed at high levels in the molecular layer of the cerebellum, although they can also been seen in the granular layer and in telencephalic regions. The present study was designed to further characterize these gacyclidine 'non-NMDA' binding sites. The pharmacological profiles obtained on cerebellar and telencephalic membrane homogenates showed that they could not be linked directly to the main receptors or uptake complexes of the central nervous system (CNS). However, the comparison of (-) and (+)[(3)H]GK11 binding distribution in different mutant animals bearing specific cellular deficits in the cerebellum has demonstrated that the gacyclidine 'non-NMDA' binding sites are associated with the dendritic trees of Purkinje cells. Interestingly, our study also shows that the radioligand binding to both cerebellar and telencephalic structures could be modulated by endogenous factors which can be removed by a stringent prewashing procedure.

Animals↗

PubMed2001

Gacyclidine: a new neuroprotective agent acting at the N-methyl-D-aspartate receptor.

Gacyclidine is a new phencyclidine derivative with neuroprotective properties. Tritiated gacyclidine and its enantiomers bind to NMDA receptors with binding parameters similar to those of other non-competitive NMDA receptor antagonists. The (-)enantiomer, (-)GK11, exhibits an affinity (2.5 nM) similar to that of dizocilpine (MK-801), while the (+)enantiomer, (+)GK11, has a 10 times lower affinity. When its interaction with NMDA receptors is prevented, gacyclidine binds also to "non-NMDA" binding sites which are mainly located in the molecular layer of the cerebellum on the dendritic tree of Purkinje cells. These binding sites do not appear to be related to any known neurotransmitters. In primary cortical cultures, gacyclidine and its enantiomers, at 0.1 to 5.0 microM, prevent glutamate-induced neuronal death. In rats, in vivo neurotoxicity of gacyclidine is far low than that of MK-801. No necrotic neurons were detected in animals sacrificed at 18 or 96 h after treatment with gacyclidine (1, 5, 10 or 20 mg/kg i.v.). At the highest (20 mg/kg) but not the lower doses (1-100 mg/kg) electron microscopy revealed the presence of few cytoplasmic or intramitochondrial vacuoles. In soman-treated monkeys gacyclidine enhanced neuroprotective activity of "three drugs cocktail" (atropine + diazepam + pralidoxime). Moreover, in rats, gacyclidine exerts a dose- and time-dependent neuroprotection in three models of spinal cord lesion. Beneficial effects of gacyclidine include reduction of lesion size and improvement of functional parameters after injury. In traumatic brain injury models gacyclidine improves also behavioral parameters and neuronal survival. Optimal protection is obtained when gacyclidine is administered at 0 to 30 min after injury. It is, therefore, concluded that gacyclidine exhibits neuroprotective effects similar to those of other NMDA receptor antagonists, with the advantage of being substantially less neurotoxic maybe due to its interaction with "non-NMDA" binding sites.

Animals↗

PubMed2000

Binding properties of [3H]gacyclidine (cis(pip/me)-1-[1-(2-thienyl)-2-methylcyclohexyl]piperidine) enantiomers in the rat central nervous system.

Gacyclidine (cis(pip/me)-1-[1-(2-thienyl)-2-methylcyclohexyl]piperidine) is a TCP derivative, which exhibits potent neuroprotective properties against glutamate-induced neurotoxicity in vitro and in vivo. In order to better understand gacyclidine pharmacological properties, the binding parameters of its enantiomers ((-) and (+)[3H]GK11) were determined in the rat central nervous system (CNS). An autoradiographic study has shown that their binding distributions are correlated with those of N-methyl-D-aspartate (NMDA) receptors throughout the CNS. Globally, the labeling was the highest with (-)[3H]GK11. In the cerebellum, both radioligands similarly labeled the molecular layer. For both radioligands, on telencephalic, cerebellum and spinal cord homogenates, the association and dissociation kinetics were accounted for by multiphasic process. In all regions, (-)[3H]GK11 exhibited the highest affinity in the nanomolar range. The pharmacological study revealed that both enantiomers labeled both high and low affinity sites in all regions. The pharmacological profile of high affinity sites was correlated with those of NMDA receptors. Those of low affinity sites were different in telencephalic and cerebellar homogenates. Overall, this study showed that low affinity sites might constitute a heterogeneous population, which could include sigma receptors in the cerebellum. The autoradiographic study has shown that these sites may be located in the molecular layer. The contribution of low affinity sites to the neuroprotective properties of gacyclidine remains to be investigated.

Animals↗

PubMed2000

Binding properties of [3H]gacyclidine in the rat central nervous system.

Gacyclidine (1-[1-(2-thienyl)-2-methylcyclohexyl]piperidine), the racemate of (+)-and (-)-GK11, exhibits potent neuroprotective properties due to its antagonism at the NMDA receptor. In its tritiated form, gacyclidine showed a binding distribution similar to that of NMDA receptors in the rat brain. With membrane preparations, the (-)-enantiomer of gacyclidine exhibited an affinity similar to that of MK-801 (dizocilpine, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a, d]cyclohepten-5,10-imine) in the low nanomolar range, while the (+)-enantiomer was about 10 times less potent. Gacyclidine affinity was lower in the cerebellum than in the forebrain or the spinal cord. In this latter region and in the cerebellum, two binding sites were evidenced, one of which was a low-affinity site insensitive to MK-801. In all regions, PRE-084 (2-(4-morpholino)ethyl-1-phenylcyclohexane-1-carboxylate), a sigma receptor ligand, had no effect on [3H]gacyclidine binding.

Animals↗

PubMed2000

The low affinity PCP sites in the rat cerebellum not only bind TCP-like but also BTCP-like structures.

Congeners of the potent dopamine (DA) re-uptake inhibitor 1-[1-(2-benzo[b]thiophenyl)cyclohexyl]piperidine (BTCP) are unexpectedly able to bind in the rat cerebellum, although this structure is devoid of dopaminergic nerve endings. In line with previous studies the hypothesis that they bind to low affinity PCP sites labelled with [3H]TCP in the rat cerebellum, even though they do not bind to the high affinity PCP sites in the forebrain, was considered. Analogues of 1-[1-(2-thiophenyl)cyclohexyl]piperidine (TCP) and BTCP with a modified aromatic moiety and with O or S atoms substituted in the cyclohexyl ring were prepared and tested in competition experiments both in rat forebrain and cerebellum membranes labelled with [3H]TCP, and in rat striatum membranes labelled with [3H]BTCP. Results indicated that BTCP and congeners could bind to low affinity PCP sites labelled with [3H]TCP in the rat cerebellum with a decrease of the selectivity for the DA transporter. On the contrary, some TCP analogues displayed a very high selectivity for these low affinity sites; they might be important pharmacological tools to elucidate the nature and function at yet unknown of these sites.

Animals↗

PubMed2000

Modulation of glutamate neurotoxicity on mesencephalic dopaminergic neurons in primary cultures by the presence of striatal target cells.

Glutamate toxicity was compared in substantia nigra (SN)/striatum (STR) and SN/cerebellum (CRB) co-cultures on both the entire neuronal population (neuron specific enolase (NSE) immunopositive cells) and dopaminergic neurons (tyrosine hydroxylase (TH) immunopositive cells). In SN/CRB co-cultures NSE- and TH-positive cells were more sensitive to glutamate-induced toxicity than in SN/STR co-cultures. Moreover, in SN/STR co-cultures as compared to SN/CRB and SN cultures, glutamate toxicity was prevented to a larger extent by TCP, a non-competitive NMDA antagonist. These results suggest that target cells induce a differential expression of the different glutamate receptor subtypes in mesencephalic dopaminergic cells. Alternatively, the presence of target cells may induce the selective development of a subpopulation of dopaminergic neurons expressing predominantly NMDA receptors.

Animals↗

PubMed2000

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) yields two active primary metabolites in vivo. Identification and quantification of BTCP primary metabolites in mice plasma, urine, and brain and their affinity for the neuronal dopamine transporter.

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) and cocaine bind to the neuronal dopamine transporter (DAT) to strongly inhibit dopamine (DA) reuptake. Although similar to acute administration, cocaine and BTCP produce sensitization and tolerance, respectively, on chronic administration. We previously found that liver microsomes produced two primary metabolites from BTCP with a high affinity for DAT. Because such metabolites, if produced in vivo, could account for the pharmacological difference with cocaine, it was important to compare BTCP biotransformations in vitro and in vivo. Therefore, we identified and quantified BTCP and primary metabolites in mice urine, plasma, and brain after acute i.p. administration. The low recovery yield suggest that BTCP might behave like its close analogue, phencyclidine, with long-term storage of metabolites. Two active metabolites found in vitro were found in mice brain with estimated half-life values similar to that of BTCP ( approximately 0.3 h). Although respective brain concentrations were 20 and 40 times lower than that of BTCP, their potency to displace in vivo [3H]BTCP bound to the DAT was 50 and 10 times higher, respectively, than that of BTCP. They could, therefore, contribute to the inhibition of DA transport and play an important role in BTCP pharmacology. They could also explain the differences between BTCP and cocaine on repeated administration.

Animals↗

PubMed1999

Does crossing over repeated treatment with the dopamine reuptake inhibitors cocaine and BTCP modify their effects on cocaine-induced locomotion?

Because the dopamine reuptake inhibitors cocaine and BTCP produce different behavioral effects after repeated administration, we studied whether they could alter each other's effects by examining the effects of crossing over repeated treatment with cocaine and BTCP on cocaine-induced locomotion. Male C57BL/6 mice were treated repeatedly with cocaine or BTCP during a first phase (days 1-3) and 3 days later, treated repeatedly with the same or the other compound during a second phase (days 7-9), after which they were administered one of several doses of cocaine on the next day. Locomotor activity was assessed after every daily treatment. The results show that 1) cocaine induced sensitization to its locomotor effects, 2) cocaine-induced sensitization was not altered by subsequent repeated treatment with BTCP, 3) initial repeated treatment with BTCP induced apparent cross-tolerance to cocaine, and 4) the initial effects of repeated BTCP were not markedly altered by subsequent repeated treatment with cocaine. The results indicate that the initial effects produced by repeated cocaine or BTCP are enduring and relatively difficult to alter by crossing over repeated treatment with the other compound. Thus, sensitization to the locomotor effects of cocaine in mice appeared to be attenuated by prior repeated treatment with BTCP but not reversed when followed by repeated treatment with BTCP.

Animals↗

PubMed1999

Pharmacological characterization of the discriminative stimulus properties of the phencyclidine analog, N-[1-(2-benzo(b)thiophenyl)-cyclohexyl]piperidine.

RATIONALE: Although both cocaine and the phencyclidine analog, BTCP, have dopamine (DA) re-uptake blocking properties, under some conditions their behavioral effects can be differentiated. Therefore, we examined whether the discriminative stimulus (DS) effects of BTCP are different from those of cocaine. OBJECTIVES: To compare the effects of monoamine re-uptake blockers, varying in their in vitro potencies as inhibitors of DA, norepinephrine (NE), or serotonin re-uptake, in different groups of rats trained to discriminate either BTCP or cocaine from saline. Additionally, drugs from other pharmacological classes were tested in both groups. METHODS: Rats were trained to discriminate either BTCP (5 mg/kg, i.p.) or cocaine (10 mg/kg, i.p.) from saline under a two-lever FR10 drug discrimination procedure. RESULTS: BTCP and cocaine cross-substituted in BTCP- and cocaine-trained rats. The DA re-uptake blockers, mazindol, indatraline, methylphenidate, GBR12909, and GBR12935, occasioned dose-related drug-lever (DL) selection both in cocaine- and in BTCP-trained rats, with potencies that were significantly correlated. In contrast, the NE re-uptake blockers, nisoxetine, desipramine, and nortriptyline, produced higher levels of DL selection in BTCP-trained rats than in cocaine-trained rats, a profile like that reported in low-dose cocaine-trained rats. Drugs from other classes acted similarly in both discriminations. Further, the alpha1-adrenergic antagonist prazosin dose dependently blocked the DS effects of the training dose of BTCP, but not of cocaine. CONCLUSIONS: The results suggest that the DS effects of BTCP are similar to cocaine, and resemble those of a low training dose of cocaine.

Animals↗

PubMed1997

1-[1-2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) yields two active primary metabolites in vitro: synthesis, identification from rat liver microsome extracts, and affinity for the neuronal dopamine transporter.

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP, 1) and cocaine bind to the neuronal dopamine transporter to inhibit dopamine (DA) reuptake. However, on chronic administration, cocaine produces sensitization, but 1 produces tolerance. Because metabolites of 1 might be responsible for some of its pharmacological properties, we have identified the primary metabolites of 1 produced by rat liver microsomes and determined their affinities for the DA transporter. Five monohydroxylated derivatives (3, 5, 9, 10, 14) and two degradation compounds (15, 16) were identified as metabolites through comparison with synthetic standards in HPLC and GC systems. Standards were obtained utilizing synthetic schemes previously used for the synthesis of phencyclidine metabolites. In vitro, two compounds (3, 5) showed a high affinity for the DA transporter. These active metabolites might be important in the pharmacology of 1.

Animals↗

PubMed1997

Long-term monitoring of extracellular dopamine concentration in the rat striatum by a repeated microdialysis procedure.

This study examined a protocol for repeated measurement of the extracellular dopamine (DA) concentration in the rat striatum by microdialysis. Rats were implanted with a guide cannula in the striatum and the probe was inserted on each dialysing day, i.e. ten times over a 23 day period. During this period the animals were submitted to a control saline treatment. DA concentration was measured using the no-net-flux method. In these conditions, DA concentration remained remarkably constant over the 23 day period. The histological analysis using glial fibrillary acidic protein (GFAP), dopamine (DA) and tyrosine hydroxylase (TH) immunocytochemistry showed a moderate gliosis and a discrete increase of immunoreactivity of catecholaminergic fibres around the probe implantation site. This increase is probably related to a plasticity of the dopaminergic system in response to the lesion due to the probe implantations. This study shows that such a paradigm makes possible to measure the whole time course of the DA concentration in the rat striatum during chronic treatments with psychoactive drugs such cocaine or other compounds acting in the nigrostriatal system.

Animals↗

PubMed1997

Comparison of glutamate and N-methyl-D-aspartate toxicities on rat mesencephalic primary cell cultures.

Excitotoxicities of glutamate and NMDA were studied on primary cultures of rat embryonic substantia nigra. The toxicity of the general neuronal population (identified with neuron specific enolase-NSE) was compared with that of dopaminergic neurons (identified with TH antibodies). We have shown that there exists a time-dependent toxicity to glutamate in 9 d old cultures in vitro and exposures as short as 5 min are significantly toxic. By comparing the effects of long time exposures (24 h) to NMDA and glutamate, we can show dose-dependent toxicity; however NMDA shows a less marked effect, especially at high doses (> 500-1000 microM) as opposed to less potent lower doses (< 500 microM). In comparison to the general population of NSE-positive mesencephalic neurons, TH-positive neurons seem to exhibit a similar vulnerability to EAA. The fact that TH-positive neurons are only partially protected against glutamate toxicity by the non-competitive NMDA antagonist TCP indicates that they are more susceptible to non-NMDA mediated neurotoxicity than the general neuronal population.

Animals↗

PubMed1997

Synthesis and biodistribution of two potential PET radioligands for dopamine reuptake sites: no-carrier-added 4-(2-[18F]fluoroethyl) and 4-[11C]methyl BTCP-piperazine.

Radioligands that specifically target dopamine uptake sites can provide a means of determining dopamine fiber loss at intrastriatal mesencephalic grafts in Parkinsonian patients, using Positron Emission Tomography (PET). The BTCP derivative, 1-[1-(2-benzo(b)thiophenyl)cyclohexyl]-4-(2-hydroxyethyl)-piperazine, shows in vitro high affinity and selectivity for the dopamine transporter. To evaluate the potential of such a compound as a potential dopaminergic PET tracer the positron-emitting analogues, 1-[1-(2-benzo(b)thiophenyl)cyclohexyl]-4-(2-[18F]fluoroethyl)-piperazine and 1-[1-(2-benzo(b)thiophenyl)cyclohexyl]-4-[11C]methylpiperazine, were synthesized. Radiofluorination was carried out by the reaction of 1-[1-(2-benzo(b)thiophenyl)cyclohexyl]-4-(2-chloroethyl)-piperazine with cyclotron-produced n.c.a. 18F-(half life 109.9 min) obtained by the (p,n) reaction on 18O-enriched water. Labelling with carbon-11 (half life 20.4 min) was achieved by 11C methylation of 1-[1-(2-benzo(b)thiophenyl)cyclohexyl]-piperazine with [11C]methyl iodide. After intravenous administration to rats these two compounds enter the brain, but despite their high in vitro affinity they display a high non specific binding in vivo which greatly limits their use as PET radioligands.

Animals↗

PubMed1996

N-[1-(2-Benzo(b)thiophenyl)cyclohexyl]piperidine (BTCP) and cocaine induce similar effects on striatal dopamine: a microdialysis study in freely moving rats.

N-[1-(2-Benzo(b)thiophenyl)cyclohexyl]piperidine (BTCP) and cocaine inhibit dopamine (DA) uptake but bind to different sites on the transporter. Their dose-dependent effects (i.p. administration) on extracellular DA levels in the rat striatum were measured by in vivo microdialysis. Both drugs dose-dependently increased DA levels with a maximum effect 60 min post injection. BTCP (20 mg/kg) had a greater peak effect than cocaine (40 mg/kg). For doses inducing similar behavioral effects (cocaine, 20mg/kg; BTCP, 10 mg/kg) similar DA increases were observed in the striatum and the nucleus accumbens. Although both drugs bind on the DA transporter on different sites and induce different behavioral effects when administered chronically, their acute administration increased striatal DA level in a similar way.

Animals↗

PubMed1996

High affinity inositol 1,3,4,5-tetrakisphosphate receptor from rat liver nuclei: purification, characterization, and amino-terminal sequence.

Inositol 1,3,4,5-tetrakisphosphate (InsP4) mediates nuclear calcium signalling [Köppler P., Matter, N., Malviya A.N. (1993) J. Biol. Chem. 268, 26248-26252], and a distinct high affinity InsP4 binding site is identified with rat liver nuclei [Köppler, P., Mersel, M., & Malviya, A.N. (1994) Biochemistry 33, 14707-14713] as compared with other rat liver membrane fractions. A novel InsP4 receptor protein derived from rat liver nuclei has been purified to apparent homogeneity employing preparative isoelectric focusing, electrophoretic mobility, nondenaturating polyacrylamide gel electrophoresis, and electroelution. Isoelectric focusing indicated an isoelectric pH around 4.3 +/- 0.2 which was further confirmed by bidimensional electrophoresis. The high affinity nuclear InsP4 receptor was identified as a 74 kDa protein both on the SDS-PAGE and on the bidimensional electrophoresis. Partial microsequence analysis showed that the N-terminal end of nuclear InsP4 receptor consists of amino acids: PNHKNEIAGNFS. The 74 kDa nuclear InsP4 receptor protein is a distinct protein from the other InsP4 receptors purified from other sources and documented in the literature.

Amino Acid Sequence↗

PubMed1996

Effects of repeated administration of N-[1-(2-benzo(b)-thiophenyl)cyclohexy]piperidine and cocaine on locomotor activity in C57BL/6 mice.

The present study investigated the effects of repeated administration of the dopamine reuptake inhibitors N-[1-(2-benzo(b)thiophenyl)cyclohexyl]piperidine (BTCP) and cocaine on locomotor activity, as well as their ability to induce or express cross-sensitization. Male mice were injected with a fixed dose of BTCP or cocaine (10,20 or 40 mg/kg i.p.) for 3 consecutive days and challenged on the 4th day with one of several doses of BTCP and/or cocaine. After every daily treatment, locomotor activity was assessed. Repeated administration of cocaine produced sensitization to the locomotor activity produced by different challenge doses of both cocaine (2.5-56.6 mg/kg i.p.) and BTCP (2.5-80 mg/kg, i.p.) on day 4. Repeated administration of low and intermediate doses of BTCP did not significantly affect the locomotor activity produced by different challenge doses of BTCP, whereas tolerance-like effects were observed after the higher dose, 40 mg/kg, particularly during the 3-day regimen. Repeated administration of BTCP dose-dependently produced leftward and downward shifts of the cocaine dose-response curve. The results indicate that, under identical treatment conditions, cocaine and BTCP differ markedly with respect to their ability to cause sensitization, but differ less in terms of their ability to elicit locomotor activity in sensitized animals. The demonstration of cross-sensitization between BTCP and cocaine provides evidence for a shared mechanism of action; however, the present results also suggest that the chronic effects of cocaine and BTCP are not identical.

Animals↗