Experiencing transitions in life of mothers of young children

V magistrskem delu smo raziskovali doživljanje življenjskih prehodov mater majhnih otrok. Teoretični del je bil namenjen pregledu literature s področja blagostanja, življenjskih prehodov in materinstva. Za raziskovanje doživljanja življenjskih prehodov je bil uporabljen psihološko fenomenološki pristop. V skladu s tem smo pridobili vpogled v doživljanje šestih udeleženk, hkrati pa vnaprej nismo predvidevali rezultatov ali postavljali hipotez. Cilj psihološko fenomenološke metode je bil oblikovati splošni opis bistvenih značilnosti doživljanja oziroma osnovne strukture, ki določa raziskovano doživljanje. Na podlagi intervjujev smo oblikovali splošno strukturo doživljanja življenjskih prehodov pri materah majhnih otrok. Ugotovili smo, da je za polovico udeleženk materinstvo najpomembnejši prehod v življenju, za ostale tri pa je na drugem mestu. Bistvene spremembe prehoda v materinstvo je mogoče razdeliti na dva dela: povzamejo občutenje, ki se dotika drugih in okolice ter občutenje, ki jih ima ženska do same sebe. Prav tako materinstvo pomembno zaznamuje doživljanje na področju telesnega, relacijskega, čustvenega in duhovnega blagostanja. Vpogled v doživljanje življenjskih prehodov pri materah majhnih otrok, ki ga daje ta raziskava, je lahko uporabna in v pomoč ljudem, ki profesionalno delujejo na področju psihologije, psihoterapije ali na katerem drugem profesionalnem področju, ki vključuje delo z ljudmi, še posebej z ženskami. Kot tako, bo lahko omogočilo pripravo strokovnih smernic programov za ženske, priprave na starševstvo ali podporne skupine za ženske. Verjamemo pa, da bo delo v pomoč tudi tistim, ki si želijo pridobiti vedenje in znanje o tej tematiki, saj lahko služi kot pomoč pri razumevanju doživljanja žensk, ko se te soočajo z življenjskimi prehodi, predvsem s prehodom v materinstvo.This Masters thesis researches how mothers with young children dealt with and experienced their life transition to the role of being a mother. The theoretical part of the thesis focuses on the literature review from the fields of welfare, life transitions and motherhood. A psychological-phenomenological approach has been used to research the life transitions. Based on this an insight into the transition experience of six women has been received, whereas there were no predictions on the outcome or setting hypothesis. The goal of the psychological-phenomenological method is to form a general description of key characteristics or the basic structure, which describe the researched transition. Based on performed interviews a general structure of experiencing the life transitions with mothers with young children has been made. The results show that for half of women experiencing motherhood is the most important transition in life, whereas the other three placed it to the second position in importance of life transitions. Key changes in the transition to the motherhood can be divided to feelings referring to the surrounding and others and secondly, feelings which a woman experiences with regard to herself. Motherhood also significantly affects woman’s relations to physical, relational, emotional and spiritual well-being. Insight to experiencing the life transitions with mothers with young children which this research offers, is useful and of help to anyone who is professionally involved in the fields of psychology, psychotherapy or any other related field which involves working with people, especially women. As such the thesis can enable preparation of professional guidelines and programs for women, parenthood preparations or support groups for women. We believe, that thesis will also assist to those, that would like to develop knowledge on this topic as it can serve as outreach at understanding of experiencing of women, when these are deal with different life transitions, particulary transition to the motherhood

Etude des mécanismes d'activation des récepteurs chimiotactiques couplés aux protéines G : Biais de signalisation et rôle d'une proline dans le deuxième domaine transmembranaire

G protein-coupled receptors (GPCRs) display a form of conservation in term ofstructure and signaling mode but also show diversification, allowing specialization. Animportant specialization during evolution is undoubtedly the pro-chemoattractant capabilitythat enables the development of organs and vascularization, or immune responses. Thesechemotactic behaviors may involve G protein couplings but also interaction with intracellularproteins including β-arrestins. However, still reside the questions on i) their ability to detectlow concentrations / molecular gradients, ii) transducing the extracellular binding domainsignal to the intracellular domain of the receptor, and iii) the modes of interaction of β-arrestins to the cytosolic GPCR domains. Recent data provided by the high-resolutionstructures indicate that GPCRs are dynamic entities that adopt multiple conformations, i.e.active able to link different types of ligands to activate in a biased manner receptor couplingpathways, leading to functional selectivity.Based on previous works of the team, we hypothesized that urotensin II (UII)-derivedpeptides displayed unexpected physiological effects because of such biased signaling on theGPCR UT receptor. We determined the coupling to G proteins and β-arrestins of the UIIactivatedUT receptor expressed in HEK293 using bioluminescence resonance energy transfer(BRET) biosensors, as well as the production of IP1-3 and cAMP using homogenous timeresolvedForster resonance energy transfer (FRET) (HTRF)-based assays. We showed thatactivated UT coupled to Gi1, GoA, Gq, and G13, excluding Gs, and recruited β-arrestins 1 and 2.Integration of these pathways led to a 2-phase kinetic phosphorylation of ERK1/2 kinases.The tested peptides induced three different profiles: UII, urotensin-related peptide (URP), andUII4–11 displayed the full profile; [Orn8]UII and [Orn5]URP activated G proteins, althoughwith pEC50s 5–10× higher, and did not or barely recruited β-arrestin; urantide also failed torecruit β-arrestin but displayed a reversed rank order of pEC50s for Gi and Gq vs. Go and was apartial agonist of all G-protein pathways. Interestingly, the peptides differently modulated cellsurvival but similarly induced cell migration and adhesion. Thus, we demonstrate biasedsignaling between β-arrestin and G proteins, and between G-protein subtypes, which dictatesthe receptor's cellular responses and may open therapeutic new opportunities.We thus addressed whether the chemotactic properties of UT, quite unexpected since theurotensinergic system was previously characterized as a potent vasoactive system, may evolvefrom a structural signature acquired early during evolution. Based on previous published data,it is proposed that an evolutionary ancestral deletion in TM2, leading to a repositioning of aAbstractproline in 2.58, of some Rhodopsine-like GPCRs belonging to the PEP (peptide) receptorfamily, allowed expansion of the SO (somatostatinergic), CHEM (chemokine) and PUR(purinergic) receptors phylogenetically linked. Thus, by studying the PEP receptor prototypeKiss1R, exhibiting a proline in 2.59, and the UT and CXCR4 receptors, as respectiveprototypes of SO and CHEM receptor groups, we aimed to gain information on the role ofthis P2.58 in the acquisition of chemotactic behaviour. Thus, we have proceeded byengineering the P2.59 prototype Kiss1R and the two P2.58 UT and CXCR4 receptors bysingle mutation of Pro to Ala or insertion/deletion to reposition the Pro.We showed that wild-type and mutant Kiss1R, UT and CXCR4 expressed in HEK293 cellsconserved expression levels and for most, binding capacities. The P2.58 UT and CXCR4receptors activated by UII and SDF-1α, exhibited Gq/Ca2+/IP1 (UT) and/or Gi/o and β-Arrestin2 couplings, as well ERK1/2 activation. They stimulated chemotactic migration via dynamicwave of assembly/disassembly of focal adhesions (FAs) associated with the selection of oneprominent lamellipodia. In contrast, activation of the P2.59 Kiss1R receptor by Kisspeptin(KP-10) evoked Gq/Ca2+/IP1 couplings, and random migration through uncontrolleddisassembly of FAs and production of many lamellipodia per cells. Single mutation orinsertion/deletion mainly led to loss of Gq and/or Gi1 and Goa coupling capacities, establishingthe key role of the proline in TM2 of Kiss1R, UT and CXCR4 in receptor activation. Inaddition, the P2.59 to P2.58 repositioning likely played a role in receptor re-sensitization,determinant for regulation of FAs assembly/disassembly and reduction of the number oflamellipodia.Thus, we propose that P2.58 receptors expanding during evolution, acquired a keycompetence to favor cell chemotactic migration under external ligand signals. This would beassociated with a dynamic of receptor desensitization/resensitization and localized adhesionde-adhesion waves, selecting protrusions necessary for cell directed migration. This specificcompetence of P2.58 in general and the UT receptor in particular, likely involves a keyfunctional selectivity towards β-arrestins.Les récepteurs couplés aux protéines G (RCPG) affichent à la fois une structure et desvoies de signalisation conservées mais aussi une grande diversité fonctionnelle, nécessaire àla spécialisation. Une spécialisation importante au cours de l'évolution est sans nul doutel’apparition du caractère chimiotactique, qui permet le développement des organes et lavascularisation, ou encore les réponses immunitaires. Ces comportements chimiotactiquesinduits par les RCPGs peuvent impliquer des couplages de protéines G mais également uneinteraction avec des protéines intracellulaires telles que les β-Arrestines. Cependant, lesquestions portant sur les RCPGs dans i) leur capacité à détecter de faiblesconcentrations/gradients moléculaires, ii) la transduction du signal depuis la poche de liaisonjusqu’au domaine intracellulaire du récepteur, et iii) les modes d'interaction des β-arrestinesrestent aujourd’hui l’objet de nombreuses recherches. Les données récentes fournies par lesstructures à haute résolution indiquent que les RCPG sont des entités dynamiques quiadoptent des conformations actives multiples, capables de lier des ligands et d’activerpréférentiellement ou non certaines voies de couplage de manière dite « biaisée », conduisantainsi à la sélectivité fonctionnelle.Sur la base de travaux antérieurs menés par l'équipe, nous avons émis l'hypothèse que lespeptides dérivés de l'urotensine II (UII) présentent des effets physiologiques inattendus enraison d'une signalisation biaisée sur le RCPG urotensinergique UT. Dans un modèle decellules HEK293 exprimant le récepteur UT humain recombinant, nous avons déterminé lecouplage aux protéines G et aux β-arrestines suite à l’activation du récepteur par l’UII, enutilisant la méthode de transfert d’énergie de bioluminescence par résonance (BRET), ainsique la production d'IP1et d'AMPc en utilisant le principe de HTR-FRET (high-time resolved-Froster resonance energy transfer). Nous avons montré que le récepteur UT active lesprotéines Gi1, GoA, Gq et G13 (et non Gs) et recrute les β-arrestines 1 et 2. Ces premiersévènements conduisent à 2 phases de phosphorylation des kinases ERK1/2. Les peptidestestés induisent trois profils différents: l'UII, l’urotensin II-related peptide (URP) et l'UII4-11présentent le profil d’agoniste entier; l’[Orn8]UII et l’[Orn5]URP activent les protéines G aveccependant des pEC50s 5 à 10 fois plus élevés, et ne recrutent pas ou peu les β-arrestines;l’urantide ne présente pas non plus la capacité d’induire le recrutement des β-arrestines mais amontré une efficacité inversement proportionnelle à activer les protéines Gi et Gq vs. Go et seprésente donc comme un agoniste partiel de toutes les voies des protéines G. Il est intéressantde noter que les peptides sont tous capables d’induire la migration et l’adhésion cellulaire,Résumémais régulent différemment la survie cellulaire. Ainsi, nous démontrons une signalisationbiaisée entre les protéines β-arrestine et G, et entre les sous-types de protéines G, ce qui dicteles réponses cellulaires du récepteur et peut ouvrir de nouvelles opportunités thérapeutiques.Nous avons ensuite examiné si les propriétés chimiotactiques de l'UT, tout à faitinattendues puisque l’UII était principalement connu pour ses propriétés vasoactives, sontassociées à une signature structurale acquise précocement au cours de l'évolution. De fait, ilavait été proposé qu'au sein des récepteurs de la famille Rhodopsine, une délétion ancestraleévolutive dans le DTM2 de récepteurs du groupe ancestral PEP (peptides), conduisant aurepositionnement d'une proline en 2.58, aurait accompagné l'expansion des récepteurs desgroupes SO (somatostatinergique), CHEM (chimiokine) et PUR (purinergique),phylogénétiquement liés. Ainsi, en étudiant le prototype de récepteur PEP Kiss1R, présentantune proline en 2.59, et les récepteurs UT et CXCR4, en tant que prototypes respectifs desrécepteurs SO et CHEM, nous avons cherché à obtenir des informations sur le rôle de cetteproline P2.58 dans l'acquisition du comportement chimiotactique. Pour cela, nous avonsgénéré des mutants des prototypes Kiss1R P2.59 et des deux récepteurs P2.58 UT et CXCR4par substitution de la Pro du DTM2 en Ala, ou par insertion/délétion pour déplacer la prolineen position 2.58 ou 2.59.Nous avons montré, dans un modèle de cellules HEK293 recombinantes, que les récepteursKiss1R, UT et CXCR4 mutants conservent une expression membranaire et pour la plupart,des capacités de liaison. Les récepteurs P2.58 UT et CXCR4 activés par l’UII et le SDF-1α,montrent des couplages Gq/Ca2+/IP1 et/ou Gi/o et β-Arrestine 2, et sont capables de stimuler lamigration chimiotactique par ondes dynamiques d'assemblage/désassemblage de pointsfocaux d’adhésion (FA), ainsi que via la sélection d’un lamellipode principal. En revanche,l'activation du récepteur Kiss1R (P2.59) par la kisspeptine-10 (KP-10) évoque des couplagesGq/Ca2+/IP1 et une migration aléatoire par désassemblage incontrôlé des FA, et la productionde nombreux lamellipodes par cellule. La mutation de la proline ou l’insertion/délétionconduit principalement à la perte des capacités de couplage Gq et/ou Gi1 et GoA, établissant lerôle clé de cette proline du DTM2 dans les mécanismes d’activation de Kiss1R, UT etCXCR4. Ainsi, le repositionnement de la proline de la position 2.59 à 2.58 joue probablementun rôle dans la re-sensibilisation des récepteurs, déterminante pour la régulation del'assemblage/désassemblage des FAs et la réduction du nombre de lamellipodes.L’ensemble de ces données nous amène à proposer que l’expansion massive des récepteursP2.58 au cours de l’évolution est corrélée à l’acquisition de mécanismes d’activation prochimiotactiquespar la régulation spatio-temporelle de signaux intracellulaires. Notamment, ceRésumémécanisme d’activation spécifique serait associé à une dynamique de désensibilisation/resensibilisationdes RCPG, à des vagues d’adhésion/détachement locales et à la sélection desprotrusions membranaires nécessaires à une migration chimiotactique efficace. Cettecompétence spécifique aux récepteurs P2.58, et en particulier de l’UT, implique probablementune sélectivité fonctionnelle clef vis-à-vis des β-Arrestines

The G Protein-Coupled Receptor UT of the Neuropeptide Urotensin II Displays Structural and Functional Chemokine Features

The urotensinergic system was previously considered as being linked to numerous physiopathological states, including atherosclerosis, heart failure, hypertension, pre-eclampsia, diabetes, renal disease, as well as brain vascular lesions. Thus, it turns out that the actions of the urotensin II (UII)/G protein-coupled receptor UT system in animal models are currently not predictive enough in regard to their effects in human clinical trials and that UII analogs, established to target UT, were not as beneficial as expected in pathological situations. Thus, many questions remain regarding the overall signaling profiles of UT leading to complex involvement in cardiovascular and inflammatory responses as well as cancer. We address the potential UT chemotactic structural and functional definition under an evolutionary angle, by the existence of a common conserved structural feature among chemokine receptorsopioïdergic receptors and UT, i.e., a specific proline position in the transmembrane domain-2 TM2 (P2.58) likely responsible for a kink helical structure that would play a key role in chemokine functions. Even if the last decade was devoted to the elucidation of the cardiovascular control by the urotensinergic system, we also attempt here to discuss the role of UII on inflammation and migration, likely providing a peptide chemokine status for UII. Indeed, our recent work established that activation of UT by a gradient concentration of UII recruits Gαi/o and Gα13 couplings in a spatiotemporal way, controlling key signaling events leading to chemotaxis. We think that this new vision of the urotensinergic system should help considering UT as a chemotactic therapeutic target in pathological situations involving cell chemoattraction

Biased signaling regulates the pleiotropic effects of the urotensin II receptor to modulate its cellular behaviors

International audienceBiased agonism by G-protein-coupled receptor ligands has opened up strategies for targeted physiological or therapeutic actions. We hypothesized that urotensin II (UII)-derived peptides displayed unexpected physiological effects because of such biased signaling on the UII human urotensin (hUT) receptor. We determined the coupling to G proteins and beta-arrestins of the UII-activated hUT receptor expressed in HEK293 using bioluminescence resonance energy transfer (BRET) biosensors, as well as the production of IP1-3 and cAMP using homogenous time-resolved Forster resonance energy transfer (FRET) (HTRF)-based assays. The activated receptor coupled to Gi1, GoA, Gq, and G13, excluding Gs, and recruited beta-arrestins 1 and 2. Integration of these pathways led to a 2-phase kinetic phosphorylation of ERK1/2 kinases. The tested peptides induced three different profiles: UII, urotensin-related peptide (URP), and UII4-11 displayed the full profile; [Orn(8)]UII and [Orn(5)]URP activated G proteins, although with pEC50s 5-10x higher, and did not or barely recruited beta-arrestin; urantide also failed to recruit beta-arrestin but displayed a reversed rank order for Gi and Gq vs. Go pEC50s (-8.79+/-0.20, -8.43+/-0.21, and -7.86+/-0.36, respectively, for urantide, -7.87+/-0.10, -7.23+/-0.27, and -8.55+/-0.19, respectively, for [Orn(5)]URP) and was a partial agonist of all G-protein pathways. Interestingly, the peptides differently modulated cell survival but similarly induced cell migration and adhesion. Thus, we demonstrate biased signaling between beta-arrestin and G proteins, and between G-protein subtypes, which dictates the receptor's cellular responses

The G Protein-Coupled Receptor UT of the Neuropeptide Urotensin II Displays Structural and Functional Chemokine Features

International audienceThe urotensinergic system was previously considered as being linked to numerous physiopathological states, including atherosclerosis, heart failure, hypertension, pre-eclampsia, diabetes, renal disease, as well as brain vascular lesions. Thus, it turns out that the actions of the urotensin II (UII)/G protein-coupled receptor UT system in animal models are currently not predictive enough in regard to their effects in human clinical trials and that UII analogs, established to target UT, were not as beneficial as expected in pathological situations. Thus, many questions remain regarding the overall signaling profiles of UT leading to complex involvement in cardiovascular and inflammatory responses as well as cancer. We address the potential UT chemotactic structural and functional definition under an evolutionary angle, by the existence of a common conserved structural feature among chemokine receptorsopioïdergic receptors and UT, i.e., a specific proline position in the transmembrane domain-2 TM2 (P2.58) likely responsible for a kink helical structure that would play a key role in chemokine functions. Even if the last decade was devoted to the elucidation of the cardiovascular control by the urotensinergic system, we also attempt here to discuss the role of UII on inflammation and migration, likely providing a peptide chemokine status for UII. Indeed, our recent work established that activation of UT by a gradient concentration of UII recruits Gαi/o and Gα13 couplings in a spatiotemporal way, controlling key signaling events leading to chemotaxis. We think that this new vision of the urotensinergic system should help considering UT as a chemotactic therapeutic target in pathological situations involving cell chemoattraction

Impact of a key ancestral deletion in chemokine and vasoactive peptide receptors on chemotaxis

International audienc

Impact of a key ancestral deletion in chemokine and vasoactive peptide receptors on chemotaxis

International audienc

The vasoactive peptide urotensin II as a new chemokine exhibits migration/adhesion mesenchymal properties accompanying angiogenesis during glioma development

National audienc

The vasoactive peptide urotensin II as a new chemokine exhibits migration/adhesion mesenchymal properties accompanying angiogenesis during glioma development

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