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Human δ opioid receptor:the effect of Phe27Cys polymorphism, N-linked glycosylation and SERCA2b interaction on receptor processing and traffickingMarkkanen, P. (Piia) 21 May 2012 (has links)
Abstract
The delta opioid receptor (δOR) is a member of the G protein-coupled receptor family. This transmembrane receptor has an important role in the regulation of pain. The OPRD1 gene that encodes the human δOR (hδOR) contains at least 11 single-nucleotide polymorphisms (SNPs). The only nonsynonymous SNP resides in the amino-terminal (N-terminal) domain of the receptor and it replaces Phe at position 27 with Cys, thus introducing an unpaired Cys residue on the extracellular surface of the receptor. The Cys27 variant has been shown to have an allelic frequency of about 10% in Caucasian populations. The polymorphic site is flanked by two putative N-glycosylation sites at Asn18 and Asn33. In this study, the folding, maturation and trafficking of hδOR was assessed using the hδORPhe27 and hδORCys27 variants and the N-glycosylation deficient forms of the latter as models in a heterologous expression system. The effects of N-glycosylation and the unpaired Cys-residue were studied with various biochemical, pharmacological and cell biological methods. In addition, protein-protein interactions of the intracellular hδOR precursors were assessed.
The hδORCys27 and hδORPhe27 variants differed significantly in their subcellular localization and maturation efficiency. The newly synthesized hδORCys27 was found to accumulate in the endoplasmic reticulum (ER) prior to its ER-associated degradation in proteasomes. Although a slow maturation rate was characteristic for both variants, only the hδORCys27 had poor maturation efficiency. The cell surface expression of hδORCys27 was further decreased because the constitutive internalization of this receptor was enhanced compared to hδORPhe27.
N-linked glycosylation was not required for hδOR function or ligand binding, but was important for the expression of the correctly folded receptor species at the cell surface. The mutant non-N-glycosylated receptor was shown to traffic to the cell surface with enhanced kinetics, but some of the plasma membrane receptors were in a nonnative conformation. Also, the overall levels of the non-N-glycosylated hδORCys27 were decreased as the receptor was efficiently internalized for lysosomal degradation in a constitutive fashion.
The hδORCys27 and hδORPhe27 precursors were found to interact with several ER localized proteins, such as calnexin (CNX), protein disulfide isomerase (PDI) and ERp72. The receptors also associated with the sarco(endo)plasmic reticulum calcium ATPase 2b (SERCA2b), which was shown to occur during translocation of the receptor to the ER membrane or immediately thereafter. The interaction was not receptor N-glycan dependent and the normal functional activity of SERCA2b was shown to be required for proper cell surface expression of hδOR. / Tiivistelmä
δ-opioidireseptori kuuluu G-proteiinikytkentäisiin reseptoreihin, ja sillä on tärkeä rooli kivun säätelyssä. Ihmisen δ-opioidireseptoria koodaavassa OPRD1 geenissä on havaittu ainakin 11 yhden nukleotidin polymorfiaa. Vain yksi tunnetuista polymorfioista aiheuttaa muutoksen proteiinin aminohapposekvenssiin. Se sijaitsee reseptorin aminoterminaalisessa osassa ja se muuttaa fenyylialaniinin (Phe) kohdassa 27 kysteiiniksi (Cys), joka on pariton. Cys27-variantin yleisyys eurooppalaisessa väestössä on noin 10 %. Polymorfisen kohdan molemmilla puolilla on N-glykosylaatiokohdat asparagiineissa Asn18 ja Asn33.
Tämän työn tavoitteena oli tutkia δ-opioidireseptorin laskostumista, maturaatiota ja kuljetusta heterologisessa solumallissa käyttämällä Phe27- ja Cys27-variantteja sekä Cys27-variantin N-glykosyloimatonta mutanttia. Cys27-polymorfian ja N-glykosylaation vaikutuksia tutkittiin useilla biokemiallisilla, farmakologisilla sekä solubiologisilla menetelmillä. Lisäksi työssä tutkittiin solunsisäisen δ-opioidireseptorin esiasteen vuorovaikutusta muiden proteiinien kanssa.
Phe27- ja Cys27-varianttien sijainti solun sisällä ja maturaatiotehokkuus eroavat toisistaan merkittävästi. Vastasyntetisoitu Cys27-variantti kerääntyy endoplasmakalvostoon, josta se ohjautuu proteasomihajoitukseen. Molemmat variantit kulkeutuvat solun pintaan hitaasti. Cys27-variantin prosessointi on huomattavasti tehottomampaa ja sen määrää solun pinnalla vähentää myös lisääntynyt ohjaaminen solunsisäiseen lysosomihajotukseen.
N-glykosylaatiolla ei havaittu olevan vaikutusta reseptorin toimintaan tai ligandin sitomiseen, mutta sillä on tärkeä merkitys oikein laskostuneiden reseptorien kuljetukselle solun pinnalle, koska osa pintaan päässeistä N-glykosyloimattomista reseptoreista on muodossa, johon reseptorispesifinen ligandi ei sitoudu. Vaikka mutanttireseptori kulkeutuukin solun pintaan nopeammin, sen määrä solun pinnalla on alhaisempi, koska mutanttireseptori ohjataan huomattavan nopeasti solun pinnalta lysosomihajotukseen.
Phe27- ja Cys27-varianttien havaittiin olevan myös vuorovaikutuksessa eräiden endosomaalisen kalvoston proteiinien kanssa, kuten kalneksiinin, proteiinidisulfidi-isomeraasin ja ERp72-proteiinin. Kumpikin reseptori havaittiin yhteisessä rakenteessa sarko(endo)plasmakalvoston kalsium-ATPaasi 2b -pumpun (SERCA2b) kanssa N-glykosylaatiosta riippumattomalla tavalla. Nämä proteiiniryhmät muodostuvat, kun reseptori liitetään synteesin aikana endoplasmakalvostoon tai heti sen jälkeen. Vuorovaikutus toiminnallisen SERCA2b:n kanssa havaittiin tärkeäksi toimintakykyisen δ-opioidireseptorin esiintymiselle solun pinnassa.
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Imagerie quantitative du traffic intracellulaire de VIH-1 / Imaging of intracellular trafficking of the HIV-1 nucleocapsid proteinLysova, Iryna 07 July 2017 (has links)
La protéine de nucléocapside du VIH-1 (NCp7) joue un rôle important dans plusieurs étapes du cycle viral. Ses interactions dans l’environnement intracellulaire dans les conditions proches d’une vraie infection sont peu décrites. L’objectif de ce projet de thèse a été de développer des approches d’imagerie permettant de suivre la NCp7 au cours des étapes précoces du cycle viral directement dans les cellules infectées. En analysant l’intensité des pseudovirus fluorescents nous avons montré un effet de « dequenching » de fluorescence témoignant d’une diminution de la concentration en NCp7 au sein des particules virales, résultant très probablement du relargage des molécules de nucléocapside à partir des complexes viraux. Ces résultats montrent pour la première fois la libération de la NCp7 dans le cytoplasme lors de la transcription inverse. Les pseudovirus NCp7-TC ont ensuite été imagés en haute résolution par microscopie PALM. Les images de distribution de la NCp7 marquée ont révélé la présence de la NCp7-TC à l'intérieur du noyau. Nous avons mis en evidence par microscopie électronique la presence de la NCp7-TC à proximité de pores nucléaires. / The nucleocapsid protein of HIV-1 (NCp7) plays an important role in several stages of the viral cycle. Its interactions in the intracellular environment under conditions close to true infection are not well described. The objective of this thesis project was to develop imaging approaches to monitor NCp7 during the early stages of the viral cycle directly in infected cells. By analyzing the intensity of the fluorescent pseudoviruses, we showed a fluorescence "dequenching" effect indicating a decrease in the NCp7 concentration within the virus particles, most likely resulting from the release of the nucleocapsid molecules from the viral complexes. These results show for the first time the release of NCp7 into the cytoplasm during reverse transcription. The pseudoviruses NCp7-TC were then imaged in high resolution by PALM microscopy. The distribution images of labeled NCp7 revealed the presence of NCp7-TC within the nucleus. We have demonstrated the presence of NCp7-TC in the vicinity of nuclear pores by electron microscopy.
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Les cellules apoptotiques vecteurs d'oncogènes viraux : Une voie alternative de la carcinogenèse associée aux HPV / Apoptotic cells as vectors of viral oncogenes : An alternative pathway of HPV-associated carcinogenesisGaiffe, Emilie 11 July 2011 (has links)
Chez les mammifères, les cellules apoptotiques peuvent être complètement dégradées par des cellules phagocytaires spécialisées ou servir de vecteur d'ADN Le transfert d'oncogènes via les cellules apoptotiques aboutit à la transformation des cellules receveuses uniquement si celles-ci sont déficientes en p53. Sachant que Fniicogène E6 des papillomavirus humains (HPV) à haut risque induit la dégradation de p53, il est concevable que son transfert par la cellule apoptotique soit à l'origine d'un mécanisme alternatif de carcinogenèse associée aux HPV. Afin de confirmer cette hypothèse, l'apoptose de cellules dérivées de cancer du col de l'utérus, abriiam ou non des séquences d'HPV, a été induite. En collaboration avec l'équipe de Patrick Sandoz du laboratoire d'optique de FEMTO-ST, nous avons adapté leur inicr;système référencé en position à l'observation automatisée de Finternalisation des cellules apoptotiques. Nous avons aussi déterminé que les cellules apoptotiques son! phagocytées par les fîbroblastes quel que soit leur statut virologique. Seules les cellules apoptotiques dérivées de cellules abritant de l'ADN d'HPV transforment les cellules receveuses. L'expression de l'ADN viral, dont E6, dans les fîbroblastes transformés ainsi que la perte d'expression des protéines p53 et p21 suggère que les oncogènes d'HPV pourraient être à l'origine de la transformation. Les résultats présentés dans ces travaux mettent en évidence un nouveau mécanisme de carcinogenèse associée aux HPV via la phagocytose des cellules apoptotiques, potentiellement impliqué dans la transformation de cellules primaires et la progression des tumeurs associées aux HPV. / Apoptotic cells in mammals may be completely degraded by specialized phagocytic cells or serve as a DNA vector. The oncogene transfer via apoptotic cells leads to the transformation of récipient cells but only when they are p53 deficient. As the E6 oncogene of high-risk human papillomavirus (HPV) leads to p53 dégradation, its transfer from apoptotic cells may be the cause of an alternative mechanism of HPV-associated carcinogenesis. To confirm this hypothesis, we induced apoptosis of cervical cancer cells that may harbor HPV sequences. In collaboration with Patrick Sandoz's team at the FEMTO-ST optical laboratory, we used their position-referenced microsystem for the automated observation of apoptotic cell internalization. We also found that apoptotic cells are phagocytized by fibroblasts regardless of their virological status. Only apoptotic cells from cells harboring HPV DNA transform recipient cells. Viral DNA expression, including E6, in transformed fibroblasts and the loss of p53 and p21 protein expression suggest that HPV oncogènes may cause transformation. These results highlight a new mechanism of HPV-associated carcinogenesis via apoptotic cell phagocytosis. This mechanism may be involved in thé transformation of primary cells and the progression of HPV-associated tumors.
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Múltiplas possibilidades: a internalização de práticas discursivo-sociais por dois pares de irmãos gêmeos / Multiple voices: the internalization of social discursive practices of two pairs of twins.Renata do Monte Vecina 28 April 2011 (has links)
Na presente pesquisa foram contempladas intersecções entre os discursos de dois pares de gêmeos, seus responsáveis e dois de seus professores sobre os modos desses gêmeos agirem, pensarem e se comportarem no mundo. Procurou-se identificar se a construção das narrativas dos gêmeos sobre os significados que atribuem à vida familiar, à vida escolar, a si próprios e à relação com seu co-gêmeo é permeada pelos discursos e práticas educativas dos pais e professores e se as práticas educativas nas quais os gêmeos se inserem e os discursos que eles escutam a respeito de si próprios podem se configurar como profecias auto-realizadoras. O discurso a respeito dos processos de desenvolvimento humano, especialmente do desenvolvimento de gêmeos, costuma seguir forte tradição determinista. A fim de romper com tradições deterministas forjadas por pressupostos endógenos ou exógenos, utilizou-se referencial teórico bakhtiniano (BAKTHIN, 2003, 2009), da psicologia histórico-cultural de Vygotsky (1984, 1993, 1997, 2000, 2004) e de autores que seguem essa linha teórica - como Oliveira (1990, 1993, 1997, 2002), Pino (2000, 2005), Rossetti-Ferreira et al. (2004, 2008, 2010), Smolka (1993, 2002, 2004), Rego (1994, 1996, 1998, 2004) - além de estabelecer debates com autores da sociologia (LAHIRE, 1997, 2002; ELIAS, 1994). O referencial teórico adotado privilegia as relações situadas social e historicamente, as significações que o indivíduo faz de si próprio e de seu mundo e a noção de sujeito discursivo e da importância das narrativas. Esta investigação visa compreender o lugar do outro nas narrativas e nas versões que os gêmeos construíram sobre si. Conhecer as expectativas de importantes agentes do processo educativo e as maneiras de apropriação desses discursos pelos gêmeos pode ajudar a reorientar modos de agir e de educar essa parcela da população. Apesar de a pesquisa ser feita especificamente com gêmeos, sua contribuição se dá também num campo mais amplo, do desenvolvimento em geral. Isso ocorre porque gêmeos são tratados como casos emblemáticos do estudo do desenvolvimento ao evidenciarem o caráter de construção da subjetividade, possibilitando o estudo de questões que acabam sendo postas para todos os indivíduos. Num âmbito maior, a pesquisa possibilita entender como as pessoas constroem versões de si próprias atravessadas pelos discursos que os outros constroem a respeito delas. A análise aponta para a importância de práticas sócio-discursivas dos outros para a constituição das versões que as pessoas fazem sobre si. A conclusão também incide sobre a necessária reflexão, por parte de pais e de professores, a respeito de seus papéis como mediadores entre seus filhos/alunos e o mundo cultural e simbólico que eles vivem, uma vez que as práticas discursivas e educativas utilizadas por eles são internalizadas por seus filhos/alunos. Constata-se, também, que os gêmeos fazem uma atualização dos lugares e dos significados sociais estabelecidos pelos pais e professores sobre seus modos de ser, de agir e de pensar e que as práticas sócio-discursivas podem, portanto, se configurar, muitas vezes, como fortes circunscritores dos processos de desenvolvimento desses jovens. / In the present study were covered intersections between discourses of two pairs of twins, their parents and two of their teachers about the ways these twins act, think and behave in the world. We sought to identify whether the construction of narratives of the twins on the meanings they attach to family life, school life, themselves and the relationship with their cotwin is permeated by the discourses and educational practices of parents and teachers and practices education in which the twins are in and the discourses they hear about themselves can be configured as self-fulfilling prophecies. Speaking about the processes of human development and especially the twin development typically follows a strong tradition of determinism. In order to break with tradition forged by deterministic assumptions endogenous or exogenous, we used theoretical Bakhtinian (BAKTHIN, 2003, 2009) and cultural-historical psychology of Vygotsky (1984, 1993, 1997, 2000, 2004) and authors who follow this theoretical line, as Oliveira (1990, 1993, 1997, 2002), Pino (2000, 2005), Rossetti-Ferreira et al. (2004, 2008, 2010), Smolka (1993, 2002, 2004), Rego (1994, 1996, 1998, 2004) and to establish discussions with authors in sociology (LAHIRE, 1997, 2002; ELIAS, 1994) theoretical framework capable of focusing on links situated socially and historically, the meanings that the individual makes of himself and his world and the notion of subject and the importance of discursive narratives. This research aims to understand the other\'s place in the narratives and versions built on the twins themselves. Knowing the expectations of important stakeholders in the educational process and ways of appropriation of these discourses by the twins can help to reorient ways and to educate this segment of the population. Although the research was done specifically with twins, its contribution is made also in a broader field of development in general. This is because twins are treated as emblematic cases of the development study which show the character of subjectivity construction, allowing the study of questions that are put to all individuals. On a wider scale, the research allows to understand how people construct versions of themselves traversed by the speeches that others built about them. The analysis points to the importance of socio-discursive practices of others to the formation of the versions that people have about themselves. The conclusion also relates to the necessary reflection of parents and teachers about their roles as mediators between their children / students and the cultural and symbolic world that they live, as the discursive practices and practices used by them are internalized by their children / students. There is relevant that the twins make an update of the \"places\" and social meanings established by parents and teachers about their ways of being, acting and thinking and that the sociodiscursive practices can therefore be set often as strong constraints of development processes of these young people.
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On the conceptual and methodological aspects of (political) journalistic translation researchKalantari, Esmaeil 05 1900 (has links)
No description available.
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Weight bias internalization, emotion dysregulation, and non-normative eating behaviors in prebariatric patientsBaldofski, Sabrina, Rudolph, Almut, Tigges, Wolgang, Herbig, Beate, Jurowich, Christian, Kaiser, Stefan, Dietrich, Arne, Hilbert, Anja January 2015 (has links)
Objective: Weight bias internalization (WBI) is associated with eating disorder psychopathology and non-normative eating behaviors among individuals with overweight and obesity, but has rarely been investigated in prebariatric patients. Based on findings demonstrating a relationship between emotion dysregulation and eating behavior, this study sought to investigate the association between WBI and eating disorder psychopathology as well as non-normative eating behaviors (i.e., food addiction, emotional eating, and eating in the absence of hunger), mediated by emotion dysregulation.
Method: Within a consecutive multicenter study, 240 prebariatric patients were assessed using self-report questionnaires. The mediating role of emotion dysregulation was examined using structural equation modeling.
Results: The analyses yielded no mediational effect of emotion dysregulation on the association between WBI and eating disorder psychopathology. However, emotion dysregulation fully mediated the associations between WBI and emotional eating as well as eating in the absence of hunger. Further, emotion dysregulation partially mediated the relationship between WBI and food addiction symptoms.
Discussion: Prebariatric patients with high levels of WBI are at risk for non-normative eating behaviors, especially if they experience emotion regulation difficulties. These findings highlight the importance of interventions targeting WBI and improving emotion regulation skills for the normalization of eating behavior in prebariatric patients.
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Obnova národa White Earth: sledování dlouhodobého procesu ústavní reformy / Rebuilding the White Earth Nation: Tracing the Long-Term Process of Constitutional ReformKrausová, Anna January 2018 (has links)
Native nation building is a phenomenon largely neglected by mainstream political science. There are empirical and theoretical gaps in the study of political structures of Native nations. The empirical focus of this dissertation is on the rebuilding process of the White Earth Nation located in northwestern Minnesota. The objective is to investigate the long-term process of White Earth governance in order to get insights into the background of the present state of the White Earth institutional stalemate. I trace external and internal factors that influenced the formation, preservation, and transformation of the White Earth government established as part of the Minnesota Chippewa Tribe under the Indian Reorganization Act provisions in 1936. To understand this process, it is necessary to include the historical context of the White Earth constitutionalism from 1913 to the present. I analyze some hitherto unknown archival materials using a flexible theoretical framework which I designed specifically for the purpose of studying the White Earth nation-building process. This case-specific framework eclectically uses a combination of theoretical approaches of Native American studies, genealogy, Vincent Pouliot's practice tracing, and new institutionalism. My findings suggest that the White Earth...
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Identification of mammalian cell signaling in response to plasma membrane perforation: Endocytosis of Listeria monocytogenes and The Repair MachineryLam, Jonathan, Lam January 2018 (has links)
No description available.
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Ligand-specific signalling at the delta opioid receptorMansour, Ahmed 12 1900 (has links)
La douleur chronique est une maladie fréquente et grave qui, pour de nombreuses personnes, ne peut pas être entièrement traitée avec les choix thérapeutiques actuels. Des agonistes des récepteurs opioïdes delta (DORs) ont été proposés comme interventions thérapeutiques pour ces maladies. Des recherches précliniques ont montré que l'activation des DOR produit des effets anti-hyperalgiques et antidépresseurs avec moins d'effets secondaires associés aux analgésiques opioïdes disponibles sur le plan clinique. Cependant, de nombreux agonistes DOR induisent une tolérance analgésique, entravant ainsi leur développement en tant que médicaments.
Les travaux de cette thèse visent à mieux comprendre les causes cellulaires et moléculaires de la tolérance ainsi que ce qui rend certains agonistes plus résistants à la tolérance que d'autres.
Dans le premier projet, nous nous sommes concentrés sur la superactivation de l'adénylyl cyclase induite par un ligand, un modèle de réponse adaptative médiée par les isoformes de l'adénylyl cyclase (AC). La superactivation de l'adénylyl cyclase (SA) a été associée à l’hyperalgésie, la tolérance analgésique et à des symptômes de sevrage. Ainsi, nous étions curieux de voir si les profils de signalisation cellulaire créés pour la découverte de médicaments pouvaient nous fournir des informations sur la capacité d'un ligand à induire la SA.
Pour répondre à cette question, nous avons généré des profils de signalisation complets pour six agonistes différents du DORs (Met-enképhaline, deltorphine II, DPDPE, SNC-80, ARM390 et TIPP) tout en surveillant 12 différents résultats de signalisation avec des biocapteurs à base de BRET. L'analyse des profils de signalisation a montré une sélectivité fonctionnelle remarquable parmi les ligands étudiés. Ensuite, nous avons pu classer les agonistes DOR en fonction de la similarité de leurs profils en utilisant l'approche que nous avons adaptée de notre laboratoire. Nous avons par la suite démontré que, à l'exception de TIPP, dont la réponse SA était Ca2+-indépendante, les catégories de médicaments résultant du regroupement sont corrélées avec la capacité du ligand à provoquer une SA. Une investigation plus approfondie des mécanismes a révélé que Gαi/o était essentiel tant pour la SA déclenchée par TIPP que par Met-Enkepkaline, mais les mécanismes en aval étaient assez distincts pour ces ligands. Ensemble, nos résultats indiquent que les mécanismes sous-jacents à la tolérance cellulaire induite par les agonistes DOR sont spécifiques au ligand.
Dans le deuxième projet, nous nous sommes principalement intéressés aux mécanismes de tolérance aux agonistes DOR qui peuvent être en partie expliqués par la désensibilisation et la régulation négative des récepteurs. Il a été établi que, les ligands qui induisent le recyclage du récepteur après l'internalisation ont été trouvés pour fournir une analgésie de longue durée. Par conséquent, les expériences menées dans cette étude ont été menées pour révéler davantage les déterminants moléculaires sous-jacents au recyclage du récepteur et sur la manière dont l'interaction agoniste-récepteur pourrait produire des modèles distincts de régulation des récepteurs.
Nous avons évalué l'activation de l'agoniste et la désensibilisation du signal DOR-Gαi1. Nos données ont rapporté que le DPDPE était pratiquement sans effet sur la désensibilisation de l'activation de Gαi1, tandis que la désensibilisation par la deltorphine II était plus importante que celle induite par le DPDPE mais moins que celle induite par l'ARM390 et le SNC-80. Ensuite, nous avons établi que les DORs stimulés par le DPDPE se recyclaient de manière plus efficace que ceux activés par la deltorphine II. De plus, nous fournissons des preuves phénoménologiques que des interventions similaires ont des effets distincts sur le recyclage évoqué par chaque ligand. En particulier, la truncation du DOR ou la surexpression de βarr2 avaient des effets différentiels sur le recyclage par le DPDPE et la deltorphine II. Il est admis que les mécanismes sous-jacents à ces différences restent à être pleinement décrits, mais la phénoménologie de nos observations soutient l'idée que le DPDPE et la deltorphine II mettent en œuvre des processus de recyclage distincts. / Chronic pain is a common and severe disease that, for many people, cannot be fully treated with current therapeutic choices. Agonists of the delta opioid receptor (DOR) have been proposed as therapeutic interventions for this illness. Preclinical research has shown that DORs produce antihyperalgesic and antidepressant-like effects with fewer side effects than the ones associated with clinically available opioid analgesics. However, numerous DOR agonists induce analgesic tolerance, hampering their development as medications. Thus, further investigations are needed to understand the mechanisms underlying the tolerance associated with chronic opioid use.
This thesis aimed to further understand the cellular and molecular mechanisms that causes tolerance as well as what makes some agonists more resistant to tolerance than others.
In the first project, we focused on ligand-induced cyclase superactivation (SA), a pattern of adaptive response mediated by adenylyl cyclase (AC) isoforms. Cyclase SA has been associated with hyperalgesia, analgesic tolerance, and withdrawal symptoms. Therefore, we were curious to assess weather cell-based signalling profiles created for drug discovery could provide us with information on the ability of a ligand to induce cyclase SA.
To address this question, we generated comprehensive signalling profiles for six different DOR agonists (Met-enkephalin, deltorphin II, DPDPE, SNC-80 and ARM390) while monitoring 12 different signalling outcomes with BRET-based biosensors.
Analysis of the signalling profiles showed remarkable functional selectivity among the investigated ligands.
Next, we were able to classify DOR agonists based on the similarity of their profiles using the approach we adapted from our lab. We subsequently demonstrated that except for TIPP, whose SA response was Ca2+-independent, the drug categories resulting from clustering are correlated with ligand capacity to cause SA.
Further investigation of the mechanisms revealed that Gαi/o was essential for both TIPP and Met-Enkepkalin-driven cyclase SA. However, downstream mechanisms were quite distinct for these two ligands. Altogether, our findings indicate that mechanisms underlying cellular tolerance induced by DOR agonists are ligand-specific.
In the second project, we were primarily concerned with the mechanisms of tolerance to DOR agonists that may be, in part, explained the receptor desensitization and downregulation. Obviously, ligands that induce receptor recycling after internalization have been found to provide long-lasting analgesia. Therefore, the objectives of the experiments in this project were to assess the molecular determinants affecting receptor recycling and how agonist-receptor interaction can result in different patterns of receptor regulation.
We assessed agonist inducing activation and desensitization of DOR-Gαi1 signal. Our data showed that DPDPE was efficient in activating the receptor without noticeable desensitization effect. On the other hand, deltorphin II exerted a significant desensitization effect. However, this effect was low when compared to ARM390 and SNC-80. Then, we established that DORs stimulated by DPDPE recycle more efficiently than those activated by deltorphin II. We also provided phenomenological evidence on receptor recycling elicited by each ligand. In particular, DOR truncation or the overexpression of βarr2 had differential effects on receptor recycling by DPDPE and deltorphin II. While our data shed light on the mechanism underlying these differences, further investigation is needed for the mechanism to be fully elucidated. Admittedly, our observations support the notion that DPDPE and deltorphin II engage distinct recycling processes.
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Les attitudes et les comportements à l’égard du corps et de l’alimentation chez des garçons adolescents : intrication de facteurs socioculturels et individuelsMorin, Guillaume 04 1900 (has links)
Thèse de doctorat présenté en vue de l'obtention du doctorat en psychologie - recherche intervention, option psychologie clinique (Ph.D) / L’adolescence est une étape importante du développement individuel lors de laquelle l’image corporelle subit des transformations impressionnantes. Chez les garçons adolescents, les difficultés liées à l’image corporelle seraient répandues et associées à plusieurs autres problématiques biopsychosociales. Néanmoins, la majorité des études réalisées à ce jour portent sur les filles. Les quelques études menées auprès de garçons omettent fréquemment la dimension musculaire de l’image corporelle, pourtant centrale chez cette population. Les recherches réalisées auprès des garçons adolescents se sont principalement appuyées sur le modèle socioculturel comme cadre théorique. Bien que plusieurs études aient permis de valider le rôle de certains facteurs socioculturels au cœur de ce modèle, peu d’entre elles se sont attardées aux facteurs individuels. Or, appuyés par de nombreuses études menées auprès de femmes, plusieurs auteurs attribuent un rôle central à certaines caractéristiques individuelles, telles que le perfectionnisme, l’estime de soi et la régulation émotionnelle, dans le développement des difficultés liées à l’image corporelle. Certains chercheurs suggèrent que les facteurs socioculturels et individuels interagissent dans l’émergence de difficultés liées à l’image corporelle. Sur la base de ces considérations, cette thèse visait à approfondir les connaissances entourant les relations entre certains facteurs socioculturels et individuels et les difficultés liées à l’image corporelle chez les garçons adolescents. Afin de répondre à cet objectif, deux études présentées sous forme d’articles ont été réalisées auprès d’un échantillon de 423 garçons âgés de 12 à 17 ans (M. = 14.7) provenant de quatre écoles secondaires de la région métropolitaine de Montréal.
La première étude visait à examiner la relation entre les pressions socioculturelles perçues liées à l’apparence, le perfectionnisme, l’estime de soi et certaines difficultés liées à l’image corporelle auprès de garçons adolescents. Elle visait aussi à explorer l’association indirecte entre ces caractéristiques individuelles et les difficultés liées à l’image corporelle par le biais des pressions socioculturelles perçues. Les difficultés liées à l’image corporelle étudiées étaient l’insatisfaction de l’image corporelle (IIC), ainsi que les attitudes et les comportements inappropriés (ACI) liés à la restriction alimentaire, à la boulimie et à la musculation. Les garçons adolescents ont rempli plusieurs questionnaires autorapportés afin d’évaluer ces difficultés liées à l’image corporelle, ainsi que les pressions socioculturelles perçues, le perfectionnisme et l’estime de soi. Les résultats suggéraient que les pressions socioculturelles perçues et le perfectionnisme, tant orienté vers soi que socialement prescrit, étaient significativement et positivement corrélés à l’IIC, ainsi qu’aux ACI liés à la restriction alimentaire, à la boulimie et à la musculation. En revanche, l’estime de soi était seulement significativement et négativement corrélée à l’IIC et aux ACI liés à la boulimie. Les résultats montraient enfin que le perfectionnisme socialement prescrit était indirectement associé à l’IIC et aux ACI liés à la restriction alimentaire, à la boulimie et à la musculation, à travers les pressions socioculturelles perçues.
La seconde étude visait dans un premier temps à examiner la relation entre l’internalisation des standards socioculturels de beauté, les difficultés de régulation émotionnelle et les difficultés liées à l’image corporelle (c.-à-d. l’IIC et les ACI liés à la restriction alimentaire, à la boulimie et à la musculation) chez un groupe de garçons adolescents. Un deuxième objectif était d’investiguer le rôle modérateur de l’internalisation des standards socioculturels de beauté dans l’association entre les difficultés de régulation émotionnelle et les difficultés liées à l’image corporelle. En plus des questionnaires précédents, les garçons adolescents ont complété des questionnaires autorapportés portant sur l’internalisation de standards socioculturels de beauté et les difficultés de régulation émotionnelle. Les résultats soutenaient que l’internalisation des standards socioculturels de beauté et les difficultés de régulation émotionnelle étaient significativement et négativement corrélées à l’ensemble des difficultés liées à l’image corporelle évaluées. Les résultats montraient que la relation entre la non-acceptation de ses émotions, l’accès limité à des stratégies de régulation émotionnelle et l’IIC était plus forte chez les garçons adolescents rapportant une internalisation plus forte de standards socioculturels de minceur. Similairement, à un degré plus élevé d’internalisation des standards socioculturels de musculature, la relation entre la non-acceptation de ses émotions, l’accès limité à des stratégies de régulation émotionnelle, l’impulsivité émotionnelle les ACI liés à la boulimie était plus forte.
Les résultats de la présente thèse appuient l’importance de s’attarder aux caractéristiques individuelles et leur intrication aux facteurs socioculturels dans l’étude des difficultés liées à l’image corporelle chez les garçons adolescents. Ces résultats vont dans le même sens que ceux obtenus dans plusieurs autres études et ils appuient certains programmes de prévention qui mettent l’emphase sur les caractéristiques socioculturelles et individuelles. Des recherches futures devront reproduire ces résultats et incorporer d’autres difficultés liées à l’image corporelle (p.ex. surveillance excessive de son apparence, l’évitement de son apparence). / Adolescence is an important stage in the human development throughout which body image undergoes extensive transformations. Body image concerns are widespread and associated with numerous biopsychosocial issues in adolescent boys. Yet, most studies until now have focused on adolescent girls. Several studies conducted among adolescent boys have omitted the muscular dimension of body image, which is central in male youth. Research conducted among adolescent boys on the topic of body image concerns has relied primarily on the sociocultural framework as their theoretical basis. Although studies have validated the role of sociocultural factors at the forefront of this model, studies focusing on individual factors are scarce. However, substantiated by the literature on women, several authors have attributed an important role to individual characteristics, such as perfectionism, self-esteem, and emotion regulation, in the development of body image difficulties. Furthermore, some researchers have argued that sociocultural and individual factors interact to promote body image concerns. Hence, this thesis aimed to increase our knowledge toward the associations between sociocultural and individual factors and body image concerns among adolescent boys. To pursue this objective, two studies presented in the form of articles were conducted in a sample of 423 boys aged from 12 to 17 years old (M. = 14.7). Participants were recruited in four high schools in Montreal metropolitan area.
The first study aimed to examine the relationship between perceived sociocultural pressures related to appearance, perfectionism, self-esteem, and body image concerns in adolescent boys. This study further aimed to explore the indirect association between these individual factors and body image concerns, through perceived sociocultural pressures. The studied body image concerns were body dissatisfaction, restrained eating, bulimic tendencies, and muscle-building strategies. Participants completed several self-reported questionnaires in order to measure their level of body image concerns, perceived sociocultural pressures, perfectionism, and self-esteem. Results suggested that perceived sociocultural pressures, self-oriented perfectionism, and socially prescribed perfectionism were positively and significantly correlated with all these body image concerns. Self-esteem was inversely and only significantly correlated with body dissatisfaction and bulimic tendencies. Results showed that socially prescribed perfectionism was indirectly associated with body dissatisfaction, restrained eating, bulimic tendencies, and muscle-building strategies through perceived sociocultural pressures.
The second study sought to investigate the relationship between the internalization of the sociocultural body ideal, difficulties in emotion regulation, and body image concerns (i.e. body dissatisfaction, restrained eating, bulimic tendencies, and muscle-building strategies) in a group of adolescent boys. A second objective was to investigate the interaction between the internalization of the sociocultural body ideal and difficulties in emotion regulation in the cross-sectional prediction of body image concerns. In addition the questionnaires described above, adolescent boys filled out self-reported inventories to assess their level of internalization of the sociocultural body ideal and difficulties in emotion regulation. Results supported that the internalization of the sociocultural body ideal and difficulties in emotion regulation were positively and significantly correlated with all these body image concerns. Results also showed that the association between non-acceptance of one’s emotion, lack of emotion regulation strategies, and body dissatisfaction was stronger in adolescent boys reporting a greater internalization of sociocultural norms of thinness. Likewise, at higher levels of internalization of sociocultural norms of muscularity, the relationship between non-acceptance of one’s emotion, lack of emotion regulation strategies, emotional impulsivity, and bulimic tendencies was stronger.
The results from our thesis support the relevance of focusing on individual characteristics and their interaction with sociocultural factors in the study of body image concerns among adolescent boys. Furthermore, these results converge with those obtained in other studies and support several prevention programs focusing on sociocultural and individual factors. Further research should replicate those findings and incorporate other body image concerns, such as body surveillance and appearance avoidance.
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