Long-Lasting Alterations in Membrane Properties, K+ Currents, and Glutamatergic Synaptic Currents of Nucleus Accumbens Medium Spiny Neurons in a Rat Model of Alcohol Dependence

Chronic alcohol exposure causes marked changes in reinforcement mechanisms and motivational state that are thought to contribute to the development of cravings and relapse during protracted withdrawal. The nucleus accumbens (NAcc) is a key structure of the mesolimbic dopaminergic reward system. Although the NAcc plays an important role in mediating alcohol-seeking behaviors, little is known about the molecular mechanisms underlying alcohol-induced neuroadaptive changes in NAcc function. The aim of this study was to investigate the effects of chronic intermittent ethanol (CIE) treatment, a rat model of alcohol withdrawal and dependence, on intrinsic electrical membrane properties and glutamatergic synaptic transmission of medium spiny neurons (MSNs) in the NAcc core during protracted withdrawal. We show that CIE treatment followed by prolonged withdrawal increased the inward rectification of MSNs observed at hyperpolarized potentials. In addition, MSNs from CIE-treated animals displayed a lower input resistance, faster action potentials (APs), and larger fast afterhyperpolarizations (fAHPs) than MSNs from vehicle-treated animals, all suggestive of increases in K+-channel conductances. Significant increases in the Cs+-sensitive inwardly rectifying K+-current accounted for the increased input resistance, while increases in the A-type K+-current accounted for the faster APs and increased fAHPs in MSNs from CIE rats. We also show that the amplitude and the conductance of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-mediated mEPSCs were enhanced in CIE-treated animals due to an increase in a small fraction of functional postsynaptic GluA2-lacking AMPARs. These long-lasting modifications of excitability and excitatory synaptic receptor function of MSNs in the NAcc core could play a critical role in the neuroadaptive changes underlying alcohol withdrawal and dependence

Les « X » à Québec : La construction discursive d’un groupe exclusif

De 1996 à 2005, l’émission radiophonique « Le monde parallèle de Jeff Fillion » prend une place prépondérante dans l’univers médiatique de Québec. L’engouement créé par cette radio repose sur un ensemble de constructions discursives qui suscitent un fort sentiment d’appartenance. Dans cet article, nous voulons explorer de façon spécifique la mécanique discursive qui contribue à la définition du groupe des X, les supporters de la station CHOI. Deux mouvements discursifs complémentaires à l’oeuvre dans la construction identitaire des X telle qu’opérée par Fillion seront décrits. Le premier mouvement, positif, consiste à proposer aux auditeurs de CHOI (des X potentiels) un rapport au monde auquel ils vont s’identifier ; le second, négatif, est une construction identitaire « contre » où l’identité des X est suggérée à travers la présentation dépréciative d’individus, de groupes et d’idéologies. De ce double mouvement émerge une représentation des X incarnant la « vraie » ville de Québec, un Québec opposé tant à celui que représente son élite, qu’au Montréal « de la ‘clique’ du Plateau Mont-Royal ». Notre étude repose sur l’analyse de cinq émissions de Fillion diffusées à l’été et à l’automne 2004, moment propice au déploiement de stratégies de polarisation alors que la survie de la station de radio CHOI est menacée et que Fillion lutte pour conserver son poste d’animateur.From 1996 to 2005, the radio program Le monde parallele de Jeff Fillion held a predominant position in Québec City’s media world. The strong following built by this radio program was generated through a series of discursive constructions that created a strong sense of belonging. This paper aims specifically to explore the discursive mechanism that contributes to the definition of Generation X, the group from which the CHOI radio station drew its audience. Two additional discursive movements that contribute to the X group’s sense of identity, as developed by Fillion, will be described. The first of these, being positive, consists in offering CHOI listeners (who are potential X’s) a relationship to the world with which they will identify; the second, which is negative, is an adverse identity construction in which the identity of the X’s is suggested through the derogatory presentation of individuals, groups, and ideologies. From this twofold movement emerges a representation of the X’s that embodies the “real” Québec City, a Québec City opposed both to that represented by its élite and to the Montréal of the “Plateau Mont-Royal clique”. Our study is based on the analysis of five broadcasts that Fillion made in the summer and autumn of 2004, an ideal moment for the deployment of polarization strategies while the survival of the CHOI radio station was threatened and Fillion was struggling to keep his position as a radio host

Adaptation de l'Archaea halophile halobacterium salinarum aux stress environnementaux : mécanismes de survie et rôle de la protéolyse intracellulaire

Molecular systems described for Archaea show primitive and simple characteristics, compared to their homologous eukaryotes. In addition, extremophilic characteristic results in an hyper-robust which makes in vitro manipulation and structural studies much easier. Thus, Archaea represent good models for understanding complex cellular functions, particularly those that involve large molecular machines, such as those involved in proteolysis. My thesis consisted in understanding the resistance mechanisms and the importance of proteolytic systems in the adaptation of halophilic Archaea to environmental stresses. Halophilic Archaea accumulate multi-molar concentrations of KCl / NaCl in their cytosol (3.4M KCl / NaCl 1.1M for Halobacterium Salinarum). This requires a very special biochemistry that allows operation in solvents where free water is scarce. Thus, the proteins of these organisms are themselves halophilic and are soluble and folded only in extreme salinity conditions (2 to 5 M). This particular biochemistry partly explain the extraordinary ability of halophilic Archaea to resist physical and chemical stress (temperature, radiation, dehydration). We study the response of the halophilic Archaea strict H. salinarum at low-salinity stress. Indeed, beyond the osmotic shock, the fall of the environment salt concentration causes a decrease in the intracellular KCl concentration, which should have a direct effect on the folding state of intracellular protein, as in case of heat stress. In the first part of this thesis, a study was conduct to determine viability limits and cytosolic modifications, associated with a salinity decrease. These studies involve intracellular salt dosages, viability studies (microscopic counts, color live / dead), induction of chaperone proteins linked to stress response and biophysical neutron experiments, to evaluate the effect of stress on proteins folding. In this work, a phenomenon of survival at low salinity linked to morphological changes was revealed. To describe this phenomenon, this second study involves confocal microscopy experiences, fluorescence microscopy, viability tests, counting on box, scanning electron microscopy, electron microscopy by negative staining, salt intracellular dosages and proteins separation experiments, to study the overall proteome composition during low salinity stress. In this study, a fall of the intracellular K⁺ concentration and the proteome clarification during stress was revealed. Low salt concentrations causes halophiles proteins denaturation, the accumulation of misfolded proteins in the cytoplasm involves chaperones systems and intracellular proteolysis machinery. In this context, another objective of my thesis was to contribute to the understanding of the intracellular proteolysis role in the PAN-proteasome system and in the aminopeptidase TET complex, in stress conditions established in previous studies. This part of the thesis involves experiments of endopeptidase activity assay, aminopeptidase activity assay, quantification of mRNA genic expression by Northern blot, immunoprecipitation, proteins separation by sucrose gradient and proteasome chemical inhibition (drug). We show the role of the PAN-proteasome system in stress response and we deepen our understanding of the aminopeptidase TET role in vivo. This protease appears to have an independent role of the proteasome complex. The protease TET seems to participate at the amino acids treatment in cells to maintain the metabolic activities in nutritional deficiencies.Les systèmes moléculaires décrits chez les Archaea mettent en évidence un caractère primitif et une simplicité, comparativement à leurs homologues eucaryotes. Par ailleurs, leur caractère extrêmophile a pour conséquence une hyper-robustesse qui rend leur manipulation in vitro et les études structurales beaucoup plus aisées. Ainsi les Archaea représentent de bons modèles pour comprendre les fonctions cellulaires complexes, particulièrement celles qui mettent en jeu de grandes machineries moléculaires, comme celles impliquées dans la protéolyse. Mon travail de thèse a consisté à comprendre les mécanismes de résistance et l'importance des systèmes de protéolyse dans l'adaptation des Archaea halophiles aux stress environnementaux. Les Archaea halophiles accumulent des concentrations multi-molaires de KCl/NaCl dans leur cytosol (3.4M KCl / 1.1M NaCl chez Halobacterium salinarum). Ainsi, les protéines de ces organismes sont elles-mêmes halophiles et ne sont solubles et repliées que dans des conditions de salinité extrêmes (de 2 à 5M).Nous avons étudié la réponse de l'Archaea halophile stricte H. salinarum à des stress, dont les stress à basse salinité, en se focalisant en particulier sur les modifications de la dynamique moléculaire du protéome in vivo (diffusion neutronique). Au cours de ce travail, il a été mis en évidence un phénomène de survie à la basse salinité associé à des modifications morphologiques.Un autre objectif de ma thèse a été de contribuer à la compréhension du rôle dans la protéolyse intracellulaire du complexe aminopeptidasique TET, dans les conditions de stress mises en place dans les études précédentes

Adaptation de l'Archaea halophile halobacterium salinarum aux stress environnementaux : mécanismes de survie et rôle de la protéolyse intracellulaire

Molecular systems described for Archaea show primitive and simple characteristics, compared to their homologous eukaryotes. In addition, extremophilic characteristic results in an hyper-robust which makes in vitro manipulation and structural studies much easier. Thus, Archaea represent good models for understanding complex cellular functions, particularly those that involve large molecular machines, such as those involved in proteolysis. My thesis consisted in understanding the resistance mechanisms and the importance of proteolytic systems in the adaptation of halophilic Archaea to environmental stresses. Halophilic Archaea accumulate multi-molar concentrations of KCl / NaCl in their cytosol (3.4M KCl / NaCl 1.1M for Halobacterium Salinarum). This requires a very special biochemistry that allows operation in solvents where free water is scarce. Thus, the proteins of these organisms are themselves halophilic and are soluble and folded only in extreme salinity conditions (2 to 5 M). This particular biochemistry partly explain the extraordinary ability of halophilic Archaea to resist physical and chemical stress (temperature, radiation, dehydration). We study the response of the halophilic Archaea strict H. salinarum at low-salinity stress. Indeed, beyond the osmotic shock, the fall of the environment salt concentration causes a decrease in the intracellular KCl concentration, which should have a direct effect on the folding state of intracellular protein, as in case of heat stress. In the first part of this thesis, a study was conduct to determine viability limits and cytosolic modifications, associated with a salinity decrease. These studies involve intracellular salt dosages, viability studies (microscopic counts, color live / dead), induction of chaperone proteins linked to stress response and biophysical neutron experiments, to evaluate the effect of stress on proteins folding. In this work, a phenomenon of survival at low salinity linked to morphological changes was revealed. To describe this phenomenon, this second study involves confocal microscopy experiences, fluorescence microscopy, viability tests, counting on box, scanning electron microscopy, electron microscopy by negative staining, salt intracellular dosages and proteins separation experiments, to study the overall proteome composition during low salinity stress. In this study, a fall of the intracellular K⁺ concentration and the proteome clarification during stress was revealed. Low salt concentrations causes halophiles proteins denaturation, the accumulation of misfolded proteins in the cytoplasm involves chaperones systems and intracellular proteolysis machinery. In this context, another objective of my thesis was to contribute to the understanding of the intracellular proteolysis role in the PAN-proteasome system and in the aminopeptidase TET complex, in stress conditions established in previous studies. This part of the thesis involves experiments of endopeptidase activity assay, aminopeptidase activity assay, quantification of mRNA genic expression by Northern blot, immunoprecipitation, proteins separation by sucrose gradient and proteasome chemical inhibition (drug). We show the role of the PAN-proteasome system in stress response and we deepen our understanding of the aminopeptidase TET role in vivo. This protease appears to have an independent role of the proteasome complex. The protease TET seems to participate at the amino acids treatment in cells to maintain the metabolic activities in nutritional deficiencies.Les systèmes moléculaires décrits chez les Archaea mettent en évidence un caractère primitif et une simplicité, comparativement à leurs homologues eucaryotes. Par ailleurs, leur caractère extrêmophile a pour conséquence une hyper-robustesse qui rend leur manipulation in vitro et les études structurales beaucoup plus aisées. Ainsi les Archaea représentent de bons modèles pour comprendre les fonctions cellulaires complexes, particulièrement celles qui mettent en jeu de grandes machineries moléculaires, comme celles impliquées dans la protéolyse. Mon travail de thèse a consisté à comprendre les mécanismes de résistance et l'importance des systèmes de protéolyse dans l'adaptation des Archaea halophiles aux stress environnementaux. Les Archaea halophiles accumulent des concentrations multi-molaires de KCl/NaCl dans leur cytosol (3.4M KCl / 1.1M NaCl chez Halobacterium salinarum). Ainsi, les protéines de ces organismes sont elles-mêmes halophiles et ne sont solubles et repliées que dans des conditions de salinité extrêmes (de 2 à 5M).Nous avons étudié la réponse de l'Archaea halophile stricte H. salinarum à des stress, dont les stress à basse salinité, en se focalisant en particulier sur les modifications de la dynamique moléculaire du protéome in vivo (diffusion neutronique). Au cours de ce travail, il a été mis en évidence un phénomène de survie à la basse salinité associé à des modifications morphologiques.Un autre objectif de ma thèse a été de contribuer à la compréhension du rôle dans la protéolyse intracellulaire du complexe aminopeptidasique TET, dans les conditions de stress mises en place dans les études précédentes

Genetic structure and evolution of the Leishmania genus in Africa and Eurasia: what does MLSA tell us

Leishmaniasis is a complex parasitic disease from a taxonomic, clinical and epidemiological point of view. The role of genetic exchanges has been questioned for over twenty years and their recent experimental demonstration along with the identification of interspecific hybrids in natura has revived this debate. After arguing that genetic exchanges were exceptional and did not contribute to Leishmania evolution, it is currently proposed that interspecific exchanges could be a major driving force for rapid adaptation to new reservoirs and vectors, expansion into new parasitic cycles and adaptation to new life conditions. To assess the existence of gene flows between species during evolution we used MLSA-based (MultiLocus Sequence Analysis) approach to analyze 222 Leishmania strains from Africa and Eurasia to accurately represent the genetic diversity of this genus. We observed a remarkable congruence of the phylogenetic signal and identified seven genetic clusters that include mainly independent lineages which are accumulating divergences without any sign of recent interspecific recombination. From a taxonomic point of view, the strong genetic structuration of the different species does not question the current classification, except for species that cause visceral forms of leishmaniasis (L. donovani, L. infantum and L. archibaldi). Although these taxa cause specific clinical forms of the disease and are maintained through different parasitic cycles, they are not clearly distinct and form a continuum, in line with the concept of species complex already suggested for this group thirty years ago. These results should have practical consequences concerning the molecular identification of parasites and the subsequent therapeutic management of the disease

Addressing the problem of plastic waste: Development of an enzymatic process for PET recycling

Every day, media and NGOs describe the society\u27s disaffection for plastics accused of polluting the planet. All major brand-owners made commitments to solve this problem (e.g. Coca-Cola, Nestlé, Danone, PepsiCo, Suntory, Unilever, L’Oréal, Nike) and announced a future with less plastic waste by 2025. Nevertheless, only 6 years before the announced term, no effective solution is yet available to meet these goals. Indeed, existing technologies like thermo-mechanical recycling leads to loss in mechanical properties of the polymer and even if several chemical recycling processes are under development, they suffer from the disadvantages of using organic solvents, high reaction temperatures and the need of an intensive waste sorting. Consequently, enzymatic recycling appears as a pertinent solution notably because the enzyme selectivity avoids a drastic sorting of waste and enables the recycling of complex plastics (multi-layers construction in some bottles of sparkling water for instance), it is an eco-friendly reaction in water and because of savings in energy consumption due to a low temperature of reaction. Using a computer-aided engineering strategy, we drastically improved the depolymerizing performance of the best identified enzyme candidate. Utilizing site-directed mutagenesis targeted at the active site, combined with three-dimensional fold stabilization, we engineered an enzyme variant, demonstrating an astounding increase in thermostability combined with a high activity. This enzyme is able to depolymerize 90% of PET waste (200g/kg) into monomers, terephthalic acid and ethylene glycol, in less than 10 hours. The downstream processing was developed and optimized leading to the demonstration that this enzymatic technology could enable the use of an industrial plastic waste to produce again PET monomers and ultimately a bottle from this recycled PET. We hope to demonstrate the strong potential of the enzymatic technology jointly developed by CARBIOS and LISBP to provide a breakthrough solution to help solve society’s growing plastic waste problem

Synthetic peripherally-restricted cannabinoid suppresses chemotherapy-induced peripheral neuropathy pain symptoms by CB1 receptor activation.

Chemotherapy-induced peripheral neuropathy (CIPN) is a severe and dose-limiting side effect of cancer treatment that affects millions of cancer survivors throughout the world and current treatment options are extremely limited by their side effects. Cannabinoids are highly effective in suppressing pain symptoms of chemotherapy-induced and other peripheral neuropathies but their widespread use is limited by central nervous system (CNS)-mediated side effects. Here, we tested one compound from a series of recently developed synthetic peripherally restricted cannabinoids (PRCBs) in a rat model of cisplatin-induced peripheral neuropathy. Results show that local or systemic administration of 4-{2-[-(1E)-1[(4-propylnaphthalen-1-yl)methylidene]-1H-inden-3-yl]ethyl}morpholine (PrNMI) dose-dependently suppressed CIPN mechanical and cold allodynia. Orally administered PrNMI also dose-dependently suppressed CIPN allodynia symptoms in both male and female rats without any CNS side effects. Co-administration with selective cannabinoid receptor subtype blockers revealed that PrNMI's anti-allodynic effects are mediated by CB1 receptor (CB1R) activation. Expression of CB2Rs was reduced in dorsal root ganglia from CIPN rats, whereas expression of CB1Rs and various endocannabinoid synthesizing and metabolizing enzymes was unaffected. Daily PrNMI treatment of CIPN rats for two weeks showed a lack of appreciable tolerance to PrNMI's anti-allodynic effects. In an operant task which reflects cerebral processing of pain, PrNMI also dose-dependently suppressed CIPN pain behaviors. Our results demonstrate that PRCBs exemplified by PrNMI may represent a viable option for the treatment of CIPN pain symptoms

Deviation from the normal mode expansion in a coupled graphene-nanomechanical system

We optomechanically measure the vibrations of a nanomechanical system made of a graphene membrane suspended on a silicon nitride nanoresonator. When probing the thermal noise of the coupled nanomechanical device, we observe a significant deviation from the normal mode expansion. It originates from the heterogeneous character of mechanical dissipation over the spatial extension of coupled eigenmodes, which violates one of the fundamental prerequisite for employing this commonly used description of the nanoresonators' thermal noise. We subsequently measure the local mechanical susceptibility and demonstrate that the fluctuation-dissipation theorem still holds and permits a proper evaluation of the thermal noise of the nanomechanical system. Since it naturally becomes delicate to ensure a good spatial homogeneity at the nanoscale, this approach is fundamental to correctly describe the thermal noise of nanomechanical systems which ultimately impact their sensing capacity

Étude expérimentale et modélisation de l'écoulement de jeu d'une aube fixe isolée

International audienceThe tip-leakage flow has detrimental effects on the performance of compressors. In this paper the effects of gap height and incoming casing boundary layer thickness are analyzed. Velocity and total pressure measurements are carried out in a plane behind the trailing edge of an isolated fixed blade. The total pressure loss is decomposed in a vortex loss and a wake loss. It appears that the increase of total pressure losses with the gap height comes essentially from the vortex part. This observation motivated the development of a model based on an analogy with a jet in crossflow to estimate the tip-leakage vortex circulation. The predictions of this model are consistent with the experimental data for gaps smaller than 4 % of chord