La métamorphose des amphibiens est le processus rapide et irréversible par lequel un têtard aquatique se transforme en une grenouille respirant à la surface. Cette transition écologique, réminiscente de la période périnatale chez les mammifères, s'accompagne de changements spectaculaires (régime alimentaire, organes locomoteurs, système respiratoire...). Ces modifications morphologiques et physiologiques nécessitent la réponse concertée à un signal hormonal, les hormones thyroïdiennes (HT), de différents tissus vers des destin parfois opposés : apoptose (dans la queue), prolifération (dans les pattes), et remodelage (dans les intestins et le système nerveux central). Toutefois, la synchronisation de la réponse des différents tissus fait appel à d'autres signaux hormonaux, et notamment les glucocorticoïdes (GC). Ces derniers sont également les médiateurs principaux de la réponse au stress. Les processus endocriniens de la métamorphose et la réponse au stress sont fortement couplés. Les GC peuvent ainsi jouer le rôle d'interface permettant l'intégration de signaux environnementaux au niveau de réseaux de régulation. Dans le cadre de mon doctorat, j'ai analysé les transcriptomes des bourgeons de membres postérieurs et de l'épiderme caudal de têtards de Xenopus tropicalis traités ponctuellement avec des HT et / ou des GC. La comparaison de ces deux tissus a permis de caractériser la diversité des profils d'expression des gènes cibles des HT et des GC.Il en ressort plusieurs résultats majeurs. Tout d'abord, la diversité des profils d'interaction entre ces deux voies est limitée, et la majorité des types de profils sont communs aux deux tissus. Indépendamment du tissu, certains profils sont caractéristiques de fonctions biologiques spécifiques comme le remodelage de la matrice extracellulaire et le système immunitaire. Les gènes impliqués dans ces fonctions communes aux deux tissus sont cependant différents. Enfin, plusieurs facteurs impliqués dans la méthylation de l'ADN sont régulés par les deux hormones. / Amphibian metamorphosis is the rapid and irreversible process during which an aquatic tadpole transforms into an air breathing adult frog. This ecological transition, reminiscent of the mammalian perinatal period, comes with spectacular changes (diet, locmotor organs, respiratory system...). These morphological and physiological modifications necessitate the properly timed response to a single hormonal signal, the thyroid hormones (TH), in various tissues to lead them to sometimes opposite fates : apoptosis (in the tail), cell prolifération and differenciation (in the limbs) and remodeling (in the intestine and the central nervous system).However, TH do not act alone. In particular, glucocorticoids (GC) play important roles during this process. They also are the main mediator of the stress response. Endocrine processes of the metamorphosis and the stress response are deeply intertwined. GC can thus act as an interface to integrate environmental inputs into regulatory networks.During my doctorate, I analyzed the possible transcriptional crosstalks between TH and GC in two larval tissues : the tailfin (TF) and the hindlimb buds (HLB). Comparing these two tissues allowed me to caracterize the diversity of TH and GC target gene expression profiles. This resulted in several major results. First, the diversity of the profiles of crosstalk between these two pathways is limited, and the majority of the types of profiles is common to both tissues. Next, independently ofthe tissues, some profiles are caracteristic of spécific biological functions such as extracellular matrix remodeling and the immune system. Yet, the genes involved in these shared functions are different between the TF and the HLB. Finally, several factors involved in DNA methylation are subject to a crosstalk between the two hormones.

A metamorfose em insetos é um dos mais complexos e belos eventos biológicos conhecidos, dirigido por sucessivas alterações morfo-fisiológicas. Este intricado processo é coordenado por componentes moleculares como ecdisteroides (20E) e hormônio juvenil (HJ), fatores de transcrição e microRNAs (miRNAs). Os miRNAs regulam a expressão de genes-alvo, que por sua vez orquestram alterações fisiológicas e anatômicas necessárias para o completo desenvolvimento do organismo. Apesar do enorme esforço, os circuitos genéticos e endócrinos que regulam a metamorfose em insetos sociais, como a abelha Apis mellifera, estão longe de serem completamente esclarecidos. Os miRNAs são importantes componentes da maquinaria celular e parecem ser ubíquos no controle de processos biológicos. Desvendar novas interações miRNA-mRNAs alvo envolvidas com a metamorfose e a regulação das cascatas de 20E e HJ lançará uma luz sobre esse complexo evento. Em nosso estudo nós investigamos os papéis de miR-34, miR-281, miR-252a e miR-252b, conhecidos como reguladores da metamorfose em insetos, no modelo A. mellifera. Todos estes miRNAs revelaram alto grau de conservação filogenética, bem como responderam ao tratamento com 20E, sofrendo flutuações na abundância de transcritos. Usando as informações disponíveis e nossos bancos de dados, nós identificamos interações envolvendo estes miRNAs e genes participantes nas cascatas de HJ e 20E: ultraspiracle (Usp), fushi tarazu-transcription factor 1 (ftz-f1), ecdysone receptor (EcR), calponin (chd64), insulin receptor 2 (inr2), e Krüppel homolog 1 (Krh1). A predição das interações miRNA-mRNAs alvo revelou que os receptores de ecdisteroides EcR e Usp, bem como o fator de transcrição ftz-f1 são alvos importantes dos miRNAs estudados, apresentando sítios para os quatros miRNAs investigados. Observamos também que os seis genes codificadores de proteína são putativamente alvejados por miR-34. Por meio do ensaio da luciferase, pudemos validar as interações entre miR-34 e os alvos Kr-h1, chd64 e inr2; miR-252a e os alvos ftz-f1 e EcR; miR-252b e os alvos chd64 e ftz-f1; miR-281 e os alvos ftz-f1, EcR e Usp. A investigação dos perfis de expressão dos miRNAs ao longo do desenvolvimento larval (L3-PP3) e pupal (Pw), contrastados com os perfis de seus respectivos alvos, apontou muitos casos de relações positivas miRNA-mRNA. Estes resultados complementaram os resultados de validação, e expuseram a regulação exercida pelo miRNA sobre seus alvos. Juntos, os nossos resultados apontam para novas interações miRNA-mRNAs, envolvidas com a metamorfose em A. mellifera. As regulações por nós propostas e validadas bem como suas caracterizações e relações com os hormônios reguladores da metamorfose, são inéditas e acrescentam muito ao conhecimento sobre a regulação da metamorfose em A. mellifera. Nesse contexto, nossa pesquisa definitivamente contribui para uma melhor compreensão dos eventos moleculares envolvidos com a metamorfose de abelhas. / Insect metamorphosis is one of the most complex and beautiful of known biological events; it consists of successive morphological and physiological alterations. This intricate process is coordinated by various molecular components, including ecdysteroids (20E), juvenile hormone (JH), transcription factors and microRNAs (miRNAs). The miRNAs regulate gene expression, which in turn orchestrates physiological and anatomical changes necessary for successful insect ontogeny. Despite enormous efforts, the endocrine and genetic circuits that regulate metamorphosis in social insects, such as honey bees (Apis mellifera), are far from being completely elucidated. The miRNAs are a substantial component of this molecular machinery and seem to be ubiquitously involved in the control of biological processes. Disclosing new miRNA-target interactions involved in metamorphosis and in the regulation of 20E and JH cascades can shed light on these poorly understood events. In this study, we provide new pieces to this puzzle. We investigated the roles of miR-34, miR-281, miR-252a and miR-252b, known to be important regulators of insect metamorphosis, in the A. mellifera model. All of these miRNAs revealed a high degree of phylogenetic conservation and responded to treatment with 20E, which altered transcript abundance. Using available information and our databases, we identified interactions involving these miRNAs and the component genes of JH and 20E pathways: ultraspiracle (Usp), fushi tarazu-transcription factor 1 (ftz-f1), ecdysone receptor (EcR), calponin (chd64), insulin receptor 2 (inr2), and Krüppel homolog 1 (Kr-h1). Prediction of miRNA-target interactions revealed that the ecdysteroid receptors EcR and Usp and the transcription factor ftz-f1 are highly targeted by miRNAs involved in metamorphosis; they presented binding sites for all four miRNAs. We also observed that all six-protein coding genes are putatively targeted by miR-34. Using the luciferase assay, we were able to validate the interactions of miR-34 with the targets Krh1, chd64 and inr2; miR-252a with the targets ftz-f1 and EcR; miR-252b with the targets chd64 and ftz-f1; and miR-281 with the targets ftz-f1, EcR and Usp. Investigation of miRNA expression profiles during larval (L3-PP3) and pupal (Pw) development, as a function of the profiles of their respective targets, demonstrated many cases of positive miRNA-mRNA relationships. These results complemented the validation results, showing how the miRNAs regulate their targets. In conclusion, we identified various previously unknown miRNA-mRNA interactions involved in the metamorphosis of A. mellifera. The regulatory pathways proposed and validated by us, as well as their characterizations and relationships with metamorphosis regulator hormones, are unique and add to the understanding of the regulation of metamorphosis in A. mellifera. In this context, our research contributes to a better understanding of the molecular events involved in honey bee metamorphosis.

No abstract available

Amphibians develop in aquatic environments where they are very susceptible to the effects of pesticides and other environmental contaminants. Glyphosate-based herbicides are widely used and have been shown to affect survival and development of tadpoles under laboratory conditions. The goal my thesis is to determine if agriculturally relevant exposure to Roundup WeatherMax®, a herbicide formulation containing the potassium salt of glyphosate and an undisclosed surfactant, influences the survival and development of wood frogs tadpoles (Lithobates sylvaticus) under both laboratory and field conditions. In the field, experimental wetlands were divided in half using an impermeable curtain so that each wetland contained a treatment and control side. Tadpoles were exposed to two pulses of this herbicide at environmentally realistic concentration (ERC, 0.21 mg acid equivalent (a.e.)/L) and predicted environmental concentrations (PEC, 2.89 mg a.e./L), after which survival, growth, development, and expression of genes involved in metamorphosis were measured. Results indicate that exposure to the PEC is extremely toxic to tadpoles under laboratory conditions but not under field conditions. Results from both experimental conditions show sublethal effects on growth and development, and demonstrate that ERC of glyphosate-based herbicides have the potential to alter hormonal responses during metamorphosis. My secondary objectives were to compare the effects of Roundup WeatherMax® to the well-studied Vision® formulation (containing the isopropylamine (IPA) salt of glyphosate and POEA), and to determine which ingredient(s) are responsible for the sublethal effects on development. Survival, growth and gene expression results indicate that Roundup WeatherMax® has greater toxicity than Vision® formulation. Contrary to my prediction, results suggest that, under realistic exposure scenarios, POEA is not the sole ingredient responsible for the observed developmental effects. However, my results demonstrate that chronic exposure to the POEA surfactant at the PEC (1.43 mg/L) is extremely toxic to wood frog tadpoles in laboratory. As part of the Long-term Experimental Wetlands Area (LEWA) project, this research contributes to overall knowledge of the impacts of glyphosate-based herbicides on aquatic communities.

Esta tesis investiga sobre el proceso de transformación de la forma arquitectónica, analizando una técnica específica denominada morphing. La técnica del morphing se utiliza en los gráficos por ordenador para la transformación de la forma entre dos o más objetos dados. Desde un punto de vista técnico, se revisan y actualizan las metodologías y aplicaciones existentes, sus características específicas y sus incidencias sobre la arquitectura. Desde un punto de vista práctico, se utilizan una serie de modelos de las casas Usonianas de Frank Lloyd Wright, con el fin de experimentar la técnica y ver qué utilidades se pueden obtener a partir de su lógica de diseño. Como resultado de este análisis se obtiene una metodología genérica para el procedimiento de un morphing arquitectónico. / This thesis investigates the transformation of architectural form, analyzing a specific technique called morphing. Morphing is a technique used in computer graphics to transform a form between two or more given objects. From a technical point of view, the existing techniques are reviewed and updated, as well as their specific characteristics and impact on architecture. From a practical point of view, some models of Usonian houses of Frank Lloyd Wright are used to experience the technique and see which utilities are available from his design logic. As a result of this analysis a generic methodology for the process of architectural morphing is obtained.

Texte remanié de : Dissertation : Sprach- und Literaturwissenschaften : Universität Erlangen-Nürnberg : 1989. / Index.

Cannibalism is a seemingly aberrant interaction, appearing counter to the fitness of individuals. Yet cannibalism is not overly uncommon, and naturally occurs among aquatic organisms, including larval amphibians. In temporary wetlands larval amphibians are in a race to complete metamorphosis before their aquatic habitat disappears. When intraspecific competition intensifies, eating conspecifics may represent a beneficial if not necessary strategy. The research presented within this thesis aims to characterize factors that influence cannibalism within populations of larval amphibians. Wood frog tadpoles (Lithobates sylvaticus) were used to test potential benefits of cannibalism as a diet, determine if dietary quality and nutritional stress influence cannibalism, and investigate the roles of competition and chemical cues in influencing cannibalism. Larval long-toed salamanders (Ambystoma macrodactylum), and ringed salamanders (A. annulatum) were used to investigate a functional link between trophic polymorphism and cannibalism in natural populations. Results suggest that perceived increases in competition may stimulate some individuals to become less risk averse, and more aggressive, which may in turn facilitate cannibalistic behaviour. Cannibalism itself provided only conditional benefits to larval wood frogs, rather than the optimal growth that would be expected from an ideal diet. However, this may have been the result of individual variation in response to the diet and/or conspecific cues as opposed to a nutritional deficit. In conditions where tadpoles could perceive increased competition they altered their behaviour and morphology in ways that may improve their foraging success and potentially promote cannibalism. Finally, a functional link appears to exist between head morphology and cannibalism in natural wetlands. However, the appearance of this morphology appears related to conditions that may facilitate increased population densities through rapid pond drying.

No description available.

Thyroid hormones (THs) are vital in the maintenance of homeostasis and in the control of development. One postembryonic developmental process that is principally regulated by THs is amphibian metamorphosis. This process has been intensively studied at the genomic level yet very little information at the proteomic level exists. There is also increasing evidence that changes in the phosphoproteome influence TH action. In addition, the disruption of TH-action by endocrine-disrupting compounds (EDC) is an emerging field and the developmental process of metamorphosis is a target as well as a model system for this research. This work identifies components of the proteome and phosphoproteome in TH-sensitive tadpole tissues that are altered during the initiation stages of TH-induced metamorphosis prior to the overt remodeling of the tissues. Proteomic analyses included two-dimensional (2D) gel electrophoresis for the assessment of differential protein/phosphoprotein expression, combined with mass spectrometry (MS) protein analysis for protein identification. Initial proteomic approaches in Xenopus laevis identified a number of proteins that are differentially expressed in the tadpole tail within 48 h of exposure of premetamorphic tadpoles to 3,5,3’-triiodothyronine (T3). Additionally, a time-course analysis of brain tissue within this 48 h period revealed alterations in phosphoproteins. The importance of phosphoproteome modulation in the process of metamorphosis was further revealed in the TH-induced tail of Rana catesbeiana tadpoles, where the inhibition of cyclin-dependent kinase activity which prevents tail regression, altered the tail phosphoproteome profile. Failure to identify the phosphoproteins involved in these initial studies led me to develop and apply new proteomic approaches. To this end, subcellular and protein fractionation methods were developed and combined with 2D gel electrophoresis and phosphoprotein-specific staining. Altered proteins were identified using MS. Here components of the proteome and phosphoproteome were identified in the tail fin that changed within 48 h of exposure of premetamorphic R. catesbeiana tadpoles to 10 nM T3. This approach allowed the identification of and led to the cloning of a novel Rana larval type I keratin, RLK I, which is a target for caspase-mediated proteolysis upon exposure to T3. In addition, the RLK I transcript level was reduced during T3-induced and natural metamorphosis, consistent with a larval keratin. Furthermore, GILT, a protein involved in the immune system, was changed in phosphorylation state which is linked to its activation. Using a complementary MS technique for the analysis of differentially-expressed proteins, isobaric tags for relative and absolute quantitation (iTRAQ) revealed 15 additional proteins whose levels were altered upon T3 treatment. The success in identifying proteins whose levels changed upon T3 treatment with iTRAQ was enhanced through de novo sequencing of MS data and homology database searching. These proteins are involved in apoptosis, extracellular matrix structure, immune system, metabolism, mechanical function, and oxygen transport. This study demonstrated the ability to derive proteomics-based information from a model species for postembryonic development for which limited genome information is currently available. The early appearance of caspase-cleaved RLK I in the TH-induced process led to its investigation as a contributor to apoptosis. Furthermore, the caspase-cleavage product of RLK I was used as a biomarker in the development of an assay for the detection of disruptors of TH-action based on ex-vivo multi-well culturing of R. catesbeiana tail fin biopsies. This assay was able to detect perturbations in TH-signalling within 48h of exposure demonstrating that it has utility as a novel system for screening of TH disrupting chemicals. The present study identified proteins whose levels and/or phosphorylation states are altered within 48 h of the induction of tadpole metamorphosis prior to overt tissue remodeling and provided important insight into the molecular mechanisms of this postembryonic development. In particular, I have identified a novel keratin that is a target for T3-mediated changes in the tail that can serve as an indicator of early response to this hormone and can be used for the detection of EDCs of TH-action in an ex vivo assay.

Thesis (doctoral)--University of Southampton. / Includes bibliographical references (p. 249-272).