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G Fillion

Publications and source records attributed to G Fillion.

At least 55 records · Page 3Linked to original sources
PubMed1996

5-hydroxytryptamine-moduline, a new endogenous cerebral peptide, controls the serotonergic activity via its specific interaction with 5-hydroxytryptamine1B/1D receptors.

The serotonergic system controls the activity of neurotransmissions involved in numerous physiological functions. It is also thought to be crucially implicated in various pathologies, including psychiatric disorders such as depression, anxiety, and aggressiveness. The properties of 5-hydroxytryptamine (5-HT)-moduline, a novel endogenous peptide, have been tested in vitro and in vivo. Binding studies have shown that the peptide specifically interacts with 5-HT1B/1D receptors via a noncompetitive mechanism corresponding to a high apparent affinity (EC50 = 10(10) M). The interaction was shown in rat and guinea pig brain tissues and in cells transfected with either 5-HT1B or 5-HT1D beta receptor gene. [3H]5-HT-moduline binds to a single population of sites in mammalian brain (Kd = 0.4 nM in rat, Kd = 0.8 nM in guinea pig) as well as in transfected cells expressing the 5-HT1B or the 5-HT1D beta receptors (Kd = 0.2 and 0.6 nM, respectively). Furthermore, the binding is clearly specific of the LSAL sequence. Autoradiographic studies showed an heterogeneous brain distribution of this site. The interaction of 5-HT-moduline with the 5-HT1B/1D receptor corresponds to a decrease in the functional activity of the receptor (i.e., a decrease in the inhibitory effect of a 5-HT1B agonist on the evoked release of [3H]5-HT from synaptosomal preparation). It was also shown that 5-HT-moduline possess an in vivo effect in the social interaction test in mouse. Finally, it was demonstrated that 5-HT-moduline was released from brain synaptosomal preparation by a K+/Ca(2+)-dependent mechanism. In conclusion, 5-HT-moduline is a novel endogenous peptide regulating the serotonergic activity via a direct action at presynaptic 5-HT receptor. It may play an important role in the physiological mechanisms involving the serotonergic system, particularly in mechanisms corresponding to the elaboration of an appropriate response of the central nervous system to a given stimulus.

Animals↗
PubMed1995

Effects of stress on the functional properties of pre- and postsynaptic 5-HT1B receptors in the rat brain.

Numerous studies have clearly shown that the turnover and release of serotonin (5-hydroxytryptamine, 5-HT) are increased under acute stressful conditions. Inasmuch as this latter process is under the control of a feedback mechanism involving the stimulation of presynaptic 5-HT1B autoreceptors, we have investigated the possible effects of acute restraint (40 min) on the functional properties of 5-HT1B receptors. The efficacy of the selective 5-HT1B receptor agonist 3-[1,2,5,6-tetrahydropyrid-4-yl]pyrrolo-[3,2-b]pyrid-5-one (CP-93,129) in inhibiting in vitro the K+-evoked release of [3H]5-HT, was significantly reduced in stressed rats as compared to naive animals. Similarly, the responsiveness of 5-HT1B receptors inhibiting the release of [3H]acetylcholine (presynaptic 5-HT1B heteroreceptors), was reduced by restraint. These effects were observed in the hippocampus, but using the inhibitory effect of CP-93,129 on forskolin-stimulated adenylyl cyclase activity as an index of 5-HT1B receptor function, it could be shown that the 5-HT1B receptors located in the substantia nigra are also desensitized by stress. The number as well as the apparent affinity constant of 5-HT1B binding sites labelled by [125I]iodocyanopindolol, as measured by quantitative autoradiography and membrane binding, were similar in naive and restraint-stressed rats suggesting that the stress-induced desensitization of 5-HT1B receptors is not due to a reduced number of 5-HT1B binding sites. As stress is thought to be a causal factor for the etiology of anxiety and depression, these results support the potential involvement of 5-HT1B receptor dysfunction in the development of these neurological disorders.

Adenylyl Cyclases↗
PubMed1995

An autoradiographic study comparing the interactions of 3 alpha-OH-5 alpha-pregnan-20-one, pregnenolone sulfate and pentobarbital with [3S]-TBPS binding sites and their modulation by GABA in different structures of the rat brain.

Using quantitative autoradiography, we have studied the distribution of the [35S]-TBPS binding sites of the GABA-A receptor complex in various structures of the rat brain. High densities of binding sites were observed in layer IV of the cerebral cortex, in the globus pallidus, and in the thalamus. Intermediate densities of binding sites were observed in superficial and deep layers of the cerebral cortex, in the dentate gyrus and in the hippocampus. For all of these structures, the interactions of 3 alpha-OH-5 alpha-pregnan-20-one (3 alpha 5 alpha P), pregnenolone sulfate (PS), and pentobarbital with [35S]-TBPS binding, in the presence or the absence of GABA were studied. In the absence of GABA, IC50 values for the inhibition of [35S]-TBPS binding were 10(-6) M for 3 alpha 5 alpha P and 10(-4) M for PS and pentobarbital in all of the brain regions studied. In the presence of GABA (5 x 10(-6) M), IC50 values were decreased by one order of magnitude for 3 alpha 5 alpha P, PS, and pentobarbital in all structures studied except layer IV of the cortex, where the IC50 value for PS was more markedly decreased (up to two orders of magnitude). By contrast, IC50 values for picrotoxin and TBPS to inhibit [35S]-TBPS binding were 10(-7) M and 10(-8) M, respectively, in the presence or absence of GABA.

Animals↗
PubMed1995

Production and characterization of an antibody directed against the mouse 5HT1B receptor.

We have developed a polyclonal antibody directed against a peptide located in the third intracellular loop of the 5HT1B receptor. Its characterization was carried out using NIH cells stably transfected with a eukaryotic expression vector containing the mouse 5HT1B receptor cDNA. The synthetic antigenic peptide had a unique sequence to the mouse 5HT1B receptor corresponding to amino acids 273-287 localized in the third intracellular loop. In dot blot analysis, antisera detected 2 ng to 2 micrograms of synthetic peptide at dilutions of 1/200-1/20,000 and bound antibody was visualized using an immunoperoxidase procedure. Preimmune serum showed no immunoreactivity to the synthetic peptide. NIH cells stably transfected expressing mouse 5HT1B receptor displayed an intense immunoreactivity with the antiserum. In contrast no immunoreactivity was seen in any of the control experiments. In the present study, we have produced a specific antibody which is an essential tool suitable for immunocytochemical applications such as regional distribution, anatomical localization and phenotypical characterization of the cells expressing the 5HT1B receptors in brain by double immunolabelling procedure.

Amino Acid Sequence↗
PubMed1995

Expression of interleukin 1 alpha, interleukin 1 beta and interleukin 1 receptor antagonist mRNA in mouse brain: regulation by bacterial lipopolysaccharide (LPS) treatment.

Lipopolysaccharide (LPS) stimulation is known to induce interleukin-1 (IL-1) mRNA expression in various immune cell types. Since IL-1 synthesis has been suggested to occur locally in brain tissue, we investigated the expression of IL-1 (alpha and beta) and IL-1 receptor antagonist (IL-1ra) mRNAs in various structures of the central nervous system, as well as in the spleen, following intraperitoneal injection of LPS (100 micrograms/mouse). After RNA extraction and amplification by the reverse transcription-polymerase chain reaction (RT-PCR), the PCR products were separated on an agarose gel, transferred and hybridized with digoxigenin-labeled probes synthetized by nested PCR. Glyceraldehyde phosphate deshydrogenase mRNA was used as an internal control. Under basal conditions the expression of IL-1 alpha, IL-1 beta and IL-1ra mRNAs in the brain was extremely low for the three cytokines; in the spleen these mRNAs were clearly detectable. Following LPS stimulation, mRNAs were strongly increased in all the tested tissues (cortex, hippocampus, hypothalamus, cerebellum, pituitary and spleen). The kinetics of mRNAs expressions in the brain were similar for all the tested regions, with a maximum at 6 h and a decrease up to 24 h after LPS administration. In the spleen the maximum was observed as soon as 1 h following stimulation. In conclusion, peripheral LPS stimulation induces a strong and transient expression of IL-1 alpha and IL-1 beta mRNAs in the brain. IL-1ra mRNA is also stimulated by LPS in various regions of the brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Stress-induced 5-HT1A receptor desensitization: protective effects of Ginkgo biloba extract (EGb 761).

The effects of sub-chronic cold stress on the functioning of hippocampal 5-HT1A receptors in old isolated rats and the possible protective effects of Ginkgo biloba extract (EGb 761) were investigated. Cold exposure during five days, produced a significant reduction of the inhibitory effect of 8-hydroxy-2-(di-n-propylamino)tetraline (8-OH-DPAT) on forskolin-stimulated adenylyl cyclase activity. In contrast, neither the affinity nor the density of hippocampal [3H]8-OH-DPAT binding sites were affected indicating that the reduced sensitivity of 5-HT1A receptors induced by stress is probably due to a modification of their coupling mechanisms to adenylyl cyclase. The stress-induced desensitization of 5-HT1A receptors was prevented by the administration of EGb 761 (50 mg/kg per os/14 days). These results clearly indicate that 5-HT1A receptors are desensitized by stress and point out the reduced capacity of old rats to cope with the adverse effects of a chronic stressor. EGb 761 appears to restore the age-related decreased capacity to adapt to a chronic stressor.

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

Rat interleukin-1 beta binding sites in rat hypothalamus and pituitary gland.

In this study, radiolabeled recombinant rat interleukin-1 beta (r125I-IL-1 beta) was used to localize and characterize IL-1 beta binding in rat hypothalamus and pituitary gland by quantitative autoradiography. The ability of this ligand to bind to type I IL-1 receptor was first tested on murine lymphoma cells (EL-4). In the rat-tissue sections, high densities of specific r125I-IL-1 beta binding sites were localized in the anterior as well as the posterior pituitary and in the choroid plexus. A fine labeling was observed in meninges and third ventricle walls while no binding was detected in the hypothalamic nuclei. Saturation experiments, in the anterior and posterior pituitary, revealed one specific binding site with an affinity constant (Kd) of 0.5 nM. Competition experiments were achieved using either rat IL-1 beta (rIL-1 beta) or human IL-1s (hIL-1 alpha, hIL-1 beta and IL-1 receptor antagonist: hIL-1a). Affinity constants (Ki) were drastically different according to the ligand used, while Ki values were found similar in anterior and posterior pituitary. Competition with rIL-1 beta revealed one binding affinity (Ki of 0.1 nM range). In contrast, competition with hIL-1 beta revealed two binding affinities: a high (Ki: 0.1 pM range) and a low one (Ki: 1 nM range). Competition with hIL-1ra was obtained for high concentrations only (Ki: 10-100 nM range), whereas human IL-1 alpha (hIL-1 alpha) was unable to compete at 1-100 nM.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Receptors for interleukin-1 in the central nervous and neuroendocrine systems. Role in infection and stress.

The functional interactions between the Immune (IS) and the Central Nervous Systems (CNS) are clearly indicated by the fact these systems are sharing mediators and receptors. Interleukins and more specifically Interleukin-1 (IL-1) have been shown to be powerful regulators of both system activity which suggested IL-1 receptors in the CNS. IL-1 receptors, similar to type I lymphocyte receptors, have been characterized in murine nervous structures (dentate gyrus of the hippocampus and frontal cortex), in vascular structures (vessels, choroid plexus) and in a neuroendocrine structure (anterior pituitary). Stimulation of the immune system and of IL-1 synthesis by bacterial product (intra peritoneal injection of LPS) induced a marked decrease of IL-1 receptor levels in the CNS. Under the same conditions pituitary receptors were unaffected indicating the autonomy of brain functioning. This decrease is in relation with an increase in local IL-1 synthesis as indicated by the increase of IL-1 mRNA in the brain tissue. During viral infection (rabies virus) very similar results are observed. Brain receptors are decreasing in the brain at day 4 post infection while IL-1 concentration is increasing in the brain tissue. Pituitary receptors are not modified during the evolution of the disease. Stress and glucocorticoid treatment are strong inhibitors of immune functions by inhibiting IL-1 synthesis. Neither treatment modified brain receptors suggesting that IL-1 synthesis is not modulated by glucocorticoids in the CNS as in the immune system. However an increase in pituitary receptor level was observed in both cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Autoradiographic characterization of binding sites for [3H]milnacipran, a new antidepressant drug, and their relationship to the serotonin transporter in rat brain.

Milnacipran is a new antidepressant drug and an equipotent inhibitor of the uptake of serotonin and noradrenaline. Quantitative autoradiography and radioligand binding studies were used to characterize recognition sites of [3H]milnacipran in rat brain. [3H]Milnacipran demonstrated saturable, reversible and nanomolar affinity binding. The binding was Na(+)-dependent, potently displaced by serotonin uptake inhibitors in all structures and moderately or weakly displaced by catecholamine uptake inhibitors (order of potency: paroxetine > fluoxetine > mazindol > desipramine > nomifensine > maprotiline). High density of recognition sites were found in structures dense in serotonergic innervation (raphe, basal ganglia, colliculi, cortex). The autoradiographic pattern of [3H]milnacipran recognition sites resembled that of [3H]paroxetine, but their distribution did not correlate well in some structures. Selective lesioning of serotonergic neurons by intracerebral injection of 5,7-dihydroxytryptamine caused a large decrease of [3H]milnacipran binding in various regions (septum, caudate, hippocampus, thalamus, ventral and dorsal hypothalamus), but in other structures, the [3H]milnacipran binding was partially affected (putamen) or even unchanged (amygdala, lateral hypothalamus). In contrast, lesion of noradrenergic neurons by intraperitoneal administration of [N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine] did not affect the binding of [3H]milnacipran in any region. These results show that [3H]milnacipran mainly binds to the serotonin transporter and does not recognize the catecholamine transporters under the conditions used. In addition, [3H]milnacipran might also bind to other sites, serotonin transporter localized on non-serotonergic neurons or serotonergic neurons insensitive to 5,7-DHT neurotoxicity.

5,7-Dihydroxytryptamine↗
PubMed1994

Characterization of galanin and 5-HT1A receptor coupling to adenylyl cyclase in discrete regions of the rat brain.

We have studied the coupling of galanin and 5-HT1A receptors with adenylyl cyclase in the hypothalamus, the entorhinal cortex and the hippocampus of the rat brain. Furthermore, we have evaluated the effects of simultaneous activation of galanin and 5-HT1A receptors on adenylyl cyclase activity. Galanin-(1-29) and galanin-(1-15) showed a dose-dependent inhibitory effect on forskolin-stimulated adenylyl cyclase activity in the hypothalamus and entorhinal cortex. No clear effects were observed in the hippocampus. Neither galanin-(1-29) nor galanin-(1-15) had any effect on the basal activity of adenylyl cyclase in these regions. The selective 5-HT1A receptor agonist 8-OH-2-(di-n-propylamino)-tetralin (8-OH-DPAT) induced a dose-dependent inhibition of forskolin stimulated adenylyl cyclase activity in the hippocampus and the entorhinal cortex. 5-HT induced an inhibition in the hypothalamus. In all regions the effects could be fully counteracted by methiothepin. 5-HT was shown to stimulate the basal activity of adenylyl cyclase in the hippocampus and the entorhinal cortex. The effects could be counteracted by methiothepin. When galanin-(1-29) and 5-HT/8-OH-DPAT were incubated simultaneously additive inhibitory effects, but no synergistic interactions, could be observed on the stimulated adenylyl cyclase activity. In conclusion, galanin and 5-HT1A receptors seem to be linked to different independent pools of G proteins, indicating that the previously demonstrated intramembrane interactions between galanin and 5-HT1A receptors involve a mechanism not directly related to adenylyl cyclase.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1994

Quantitative autoradiography of 5-HT1D and 5-HT1E binding sites labelled by [3H]5-HT, in frontal cortex and the hippocampal region of the human brain.

In human cortex and hippocampus area, [3H]5-HT (5 nM) labels 5-HT1A, 5-HT1D and 5-HT1E sites. After masking 5-HT1A receptors by 0.1 microM 8-OH-DPAT, the binding displaced by 0.1 microM 5-CT presumably represented 5-HT1D sites and the remaining binding 5-HT1E sites. In frontal cortex, 5-HT1A receptors represented the main binding in layers II and VI and a lower fraction in other layers. 5-HT1D and 5-HT1E sites, were more homogeneously distributed in layers II to VI (21-34% of specific [3H]5-HT binding). 5-HT1E sites were of similar affinities (KD close to 6-8 nM) in the cortical layers II to VI. In CA1 field of hippocampus, (pyramidal layer, stratum radiatum, molecular layer), CA2 and dentate gyrus, 5-HT1A receptors represented the major fraction, 5-HT1D sites a significant fraction and 5-HT1E a minor fraction of the specific [3H]5-HT binding. In CA3-CA4 fields, 5-HT1A receptors were less densely present, 5-HT1D sites were predominant and 5-HT1E sites represented a significant fraction (27%). The highest densities of 5-HT1E sites have been measured in subiculum, where 5-HT1A, 5-HT1D and 5-HT1E binding sites were equally represented and in entorhinal cortex where 5-HT1E sites represented the major binding in layer III. They were also present in layers II and IV (29 and 24%) and, to a lesser extent, in layers V and VI. 5-HT1A sites were predominant in layer VI, II and V and were less abundant in other layers. 5-HT1D were homogeneously present in layers II, III, IV and were present in low amounts in other layers. No 5-HT1E were detected in choroid plexus, where [3H]5-HT was dramatically reduced by mesulergine (5-HT2C receptors). No significant displacement of [3H]5-HT by mesulergine was measured in other structures.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1994

Effect of chronic antidepressant treatment on 5-HT1B presynaptic heteroreceptors inhibiting acetylcholine release.

The effect of long term treatment with two tricyclic antidepressants on the sensitivity of 5-HT1B presynaptic heteroreceptors inhibiting acetylcholine (ACh) release was investigated. Groups of male rats received during 14 days either saline, citalopram (20 mg/kg), a serotonin (5-HT) uptake blocker, or tianeptine (2 x 10 mg/kg), an antidepressant that enhances 5-HT uptake. The efficacy of the 5-HT1B selective agonist 7-trifluoromethyl-4-(4-1-piperazinyl)-pyrrolo[1,2-a]quinoxaline (CGS 12066B) in reducing K(+)-evoked [3H]acetylcholine release from hippocampal synaptosomes was determined 24 hr after the last administration. The chronic treatment with citalopram or tianeptine modified neither the basal nor the K(+)-evoked release of [3H]acetylcholine. In contrast, these treatments significantly reduced the efficacy of CGS 12066B to inhibit the release of [3H]acetylcholine induced by K+ depolarization. These data suggest that chronic antidepressant treatment desensitizes 5-HT1B presynaptic heteroreceptors through a mechanism which seems to be independent of the synaptic availability of 5-HT.

Acetylcholine↗
PubMed1994

"In situ" characterization of GnRH receptors: use of two radioimagers and comparison with quantitative autoradiography.

New radioimagers, the HRRI (high resolution radioimager) and the Phosphorimager (phosphor screen : PS), apt to display more ample linear dose-response scale than radio-sensitive films, were tested in comparison with quantitative autoradiography (QA). GnRH receptor saturation experiments were achieved on tissue sections (rat pituitary, rat brain, human ovary) with a iodinate GnRH agonist (125I-[D-Ala6,Des-Gly10]-LH-RH Ethylamide) for determination of affinity constant (Kd). In rat pituitary, comparable results were obtained with the 3 methods (Kd: 0.4 to 0.6 nM). Discrepancies occurred in the hippocampus and in the granulosa cell layer of the preovulatory follicle, due to low resolutive (PS) or short linear dose-response (films) performances. In the hippocampus GnRH receptor affinity was under-estimated with PS (Kd: 2.3 vs 0.5 and 0.6 nM for QA and HRRI respectively). In the follicular granulosa cell layer it was over-estimated by QA (0.5 vs 50 nM for the HRRI), while PS did not allow resolution of this thin cell layer. In conclusion, the HRRI is a very powerful tool for the quantification of in situ radioligand binding (binding sites study and in situ hybridization) in very discrete areas.

Adult↗
PubMed1993

Quantitative autoradiography of 5-HT1E binding sites in rodent brains: effect of lesion of serotonergic neurones.

Binding sites corresponding to 5-HT1E receptors were labelled in mouse, rat, and guinea-pig brains by using [3H]5-hydroxytryptamine ([3H]5-HT) in the presence of 5-carboxamidotryptamine (5-CT) (0.1 microM), and their distribution within the brain was studied by quantitative autoradiography. The results obtained with mouse brain show that 5-HT1E binding sites are particularly present in the cortex, caudate-putamen and claustrum, where they showed the highest density. Lower densities were measured in other regions. Saturation experiments showed that the affinity of [3H]5-HT for 5-HT1E binding sites (nanomolar range) was very similar in the different structures. The distribution of 5-HT1E binding sites was similar in rat and guinea-pig brains. In rat brain, selective lesioning of serotonergic fibres by intracerebroventricular injection of 5,7-dihydroxytryptamine (5,7-DHT), a specific 5-HT neurotoxin, did not affect the density of 5-HT1E binding, indicating that these receptors are mainly localized on non-serotonergic neurones.

5,7-Dihydroxytryptamine↗