Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 74-81).

No description available.

Formation of segments during vertebrate embryogenesis is regulated by a biological clock. Models and experimental data indicate that the core of this clock consists of a cell- autonomous single cell oscillator. This oscillator likely involves a genetic feedback loop of transcriptional repressors belonging to the hairy gene family. In zebrafish, three her genes, her1, hes6 and her7, have been identified as core oscillator components. The main purpose of this project was to study the molecular mechanism of the hairy gene negative feedback oscillator in single cells. To determine whether a single cell oscillator is part of the zebrafish segmentation clock, a cell dissociation protocol was established to track the expression of Her1 ex vivo. Upon dissociation, Her1 expression continued to oscillate for up to three cycles. The period of oscillations was significantly slower than that of the segmentation clock, but appears to speed up in the presence of serum. To test whether the hairy gene interactions are sufficient to generate oscillations in single cells, a protocol was established that uses synthetic biology principles to design, construct and characterize hairy gene networks in yeast. First a library of network parts, containing hairy genes, promoters and Her binding sites was generated and subsequently assembled into simple devices to test their functionality in yeast. The three core oscillator components, Her1, Hes6 and Her7, were characterized and optimized for expression in yeast. In the SWITCH-OFF assay, the Her1 protein, modified with a MigED yeast repressor domain, was found to function as a transcriptional repressor in yeast, while Hes6 with the same modification can not. The dissociation of segmentation clock cells provides the first direct evidence that single cell oscillators exist in zebrafish. In this system, oscillator dynamics can be studied without the interactions of higher level clock components. In parallel, establishing a yeast chassis for hairy gene networks provides a novel technique to directly test predicted oscillator mechanisms by constructing them ’bottom up’.

Generation of the primitive neuroectoderm into specialized brain subdivisions, such as the hindbrain primordium, involves the regulated coordination of complex morphogenetic and molecular mechanisms. These processes are evident in the segregation of the zebrafish hindbrain into seven distinct lineage-restricted compartments, termed rhombomeres (r), which are established by the interplay of several spatially-restricted expressed genes. These include transcription factors, members of specific signaling pathways and specialized molecules that mediate cell adhesion and identity. Despite their extensive characterization, it is evident that other genes are involved to mediate the proper specification and segregation of individual rhombomeres. One candidate that likely fits this role is related to the no ocelli/l(2)35Ba gene in Drosophila, termed nlz (nocA-like zinc-finger). Nlz-related proteins behave as transcriptional repressors and are related to the vertebrate Sp1-like family of transcription factors. nlz is dynamically expressed in the zebrafish hindbrain, residing in the caudal hindbrain at gastrula stages and rostrally expanding from presumptive r3/r4 boundary to encompass r3 and r2 at segmentation stages. Nlz localizes to the nucleus and associates with the co-repressors Groucho and histone deacetylases, suggesting that Nlz acts as a repressor. Consistent with this, misexpression of nlz into zebrafish embryos results in a loss of gene expression in the rostral hindbrain (rl-r3). Taken together, the findings in this thesis suggest that Nlz functions as a transcriptional repressor to control segmental gene expression in the rostral hindbrain.

A cetamina é uma droga amplamente utilizada e o seu uso inadequado tem sido associado à graves consequências para a saúde humana. Embora as propriedades farmacológicas deste agente em doses terapêuticas sejam bem conhecidas, existem poucos estudos sobre os efeitos secundários induzidos por doses não-terapêuticas, incluindo os efeitos nos estados de ansiedade e agressividade. Neste contexto, os modelos animais são uma etapa importante na investigação e elucidação do mecanismo de ação a nível comportamental. O zebrafish (Danio rerio) é um novo organismo-modelo, interessante e promissor, uma vez que apresenta alta similaridade fisiológica, genética e neuroquímica com seres humanos, respostas comportamentais bem definidas e rápida absorção de compostos de interesse em meio aquoso além de apresentar uma série de vantagens em relação aos modelos mamíferos tais como manutenção de baixo custo, prática e executável em espaços reduzidos. Nesse sentido, faz-se necessário a execução de ensaios comportamentais em conjunto com análises estatísticas robustas e rápidas tais como ANOVA e Métodos Multivariados; e também o desenvolvimento de métodos analíticos sensíveis, precisos e rápidos para determinação de compostos de interesse em matrizes biológicas provenientes do animal. Os objetivos do presente trabalho foram a investigação dos efeitos da cetamina sobre a ansiedade e a agressividade em zebrafish adulto empregando Testes de Claro-Escuro e Testes do Espelho e métodos estatísticos univariados (ANOVA) e multivariados (PCA, HCA e SIMCA) assim como o desenvolvimento de método analítico para determinação da cetamina em matriz biológica proveniente do animal, empregando Extração Líquido-Líquido e Cromatografia em Fase Gasosa acoplada ao Detector de Nitrogênio-Fósforo (GC-NPD). Os resultados comportamentais indicaram que a cetamina produziu um efeito significativo dose-dependente em zebrafish adulto na latência à área clara, no número de cruzamentos entre as áreas e no tempo de exploração da área clara. Os resultados das análises SIMCA e PCA mostraram uma maior similaridade entre o grupo controle e os grupos de tratamento expostos às doses mais baixas (5 e 20 mg L-1) e entre os grupos expostos às doses de 40 e 60 mg L-1. Na análise por PCA, dois componentes principais responderam por 88,74% de toda a informação do sistema, sendo que 62,59% da informação cumulativa do sistema foi descrito pela primeira componente principal. As classificações HCA e SIMCA seguiram uma evolução lógica na distribuição das amostras por classes. As doses mais altas de cetamina induziram uma distribuição mais homogênea das amostras enquanto as doses mais baixas e o controle resultaram em distribuições mais dispersas. No Teste do Espelho, a cetamina não induziu efeitos significativos no comportamento dos animais. Estes resultados sugerem que a cetamina é modulador de comportamentos ansiosos, sem efeitos indutores de agressividade. Os resultados da validação do método cromatográfico indicaram uma extração com valores de recuperação entre 33,65% e 70,89%. A curva de calibração foi linear com valor de R2 superior a 0,99. O limite de detecção (LOD) foi de 1 ng e o limite de quantificação (LOQ) foi de 5 ng. A exatidão do método cromatográfico manteve-se entre - 24,83% e - 1,258%, a precisão intra-ensaio entre 2,67 e 14,5% e a precisão inter-ensaio entre 1,93 e 13,9%. / Ketamine is a widely used drug and its inappropriate use has been associated with serious consequences for human health. Although the pharmacological properties of this agent in therapeutic doses are well known, there are few studies about the side effects induced at non-therapeutic doses, including the effects on states of anxiety and aggression. In this context, animal models are an important step in the investigation and elucidation of the mechanism of action at the behavioral level. The zebrafish (Danio rerio) is a new organism model, interesting and promising, since it presents high physiological, genetics and neurochemistry similarity in relation to humans, well-defined behavioral responses and rapid absorption of interesting compounds in an aqueous medium, apart from presents several advantages over mammalian models such as practical, low cost maintenance and executable in reduced spaces. In this sense, it is necessary to perform behavioral tests in conjunction with robust and rapid statistical analysis such as ANOVA and multivariate methods; and also the development of sensitive, accurate and rapid analytical methods to determination of compounds of interest in biological matrices from the animal. The objectives of this study were to investigate the effects of ketamine on anxiety and aggression in adult zebrafish using Light-Dark and Mirror Biting tests and univariate (ANOVA) and multivariate (PCA, HCA and SIMCA) statistical methods and to develop an analytical method for determination of ketamine in biological animal matrices using Liquid-Liquid Extraction and Gas Chromatography coupled with Nitrogen-Phosphorus Detector (GC-NPD). The behavioral results of Light-Dark Test indicated that ketamine produced a significant dose-dependent response in the latency to light area, in the number of midline crossings and in the time spent in light area. Results of SIMCA and PCA analysis showed a greater similarity between the control group and treatment groups exposed to lower doses (5 and 20 mg L-1) and between the treatment groups at doses of 40 and 60 mg L-1. In the analysis PCA, two principal components accounted for 88,74% of all the system information and 62,59% of the cumulative information of the system were described for the first principal component. The HCA and SIMCA results showed a logical evolution in the distribution of samples per class. The higher dose of ketamine induced a more homogeneous distribution of the samples while the lower doses and control resulted in more dispersed distribution. In the Mirror Test, ketamine induced no significant effect on the behavior of animals. These results suggest that ketamine is a modulator of anxious behavior without inducing aggressive effects. The results of the validation of chromatographic method indicated an extraction with recovery ranged between 33,65% to 70,89%. Calibration curve was linear with R2 value higher than 0,99. The limit of detection (LOD) was 1 ng and the limit of quantification (LOQ) was 5 ng. The accuracy of gas chromatographic method ranged between - 24,83% and - 1,258%, intra-assay precision between 2,67 and 14,5% and inter-assay precision between 1,93 and 13,9%.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2018-05-17T18:37:10Z No. of bitstreams: 0 / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2018-05-22T11:40:51Z (GMT) No. of bitstreams: 0 / Made available in DSpace on 2018-05-22T11:40:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-07-27 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A artepilina C é o principal marcador químico utilizado no controle de qualidade da própolis verde, a qual é produzida especialmente em Minas Gerais e comercializada principalmente para o Japão e que possui como principal fonte botânica a planta Baccharis dracunculifolia DC. (Asteraceae). Entre as técnicas que podem ser utilizadas para a análise da artepilina C, a cromatografia líquida de ultra eficiência (CLUE) acoplada a espectrometria de massas (EM) apresenta inúmeras vantagens. Em relação às suas atividades biológicas, a artepilina C apresenta ação antioxidante e antitumoral, não tendo sido ainda avaliada sua embriotoxicidade e atividade antienvelhecimento. O presente trabalho teve como objetivos isolar a artepilina C, desenvolver e validar método analítico de quantificação dessa substância por CLUE-EM-Q-TOF, bem como determinar sua atividade antienvelhecimento em nematoides Caenorhabditis elegans e sua toxicidade em embriões de Zebrafish. Para isolar a artepilina C de amostra de própolis verde, foi realizada extração básica seguida de acidificação, com posterior fracionamento por cromatografia líquida a vácuo (CLV) e cromatografia líquida de alta eficiência (CLAE) semipreparativa. Para identificação da substância, foi realizada análise por CLAE e ressonância magnética nuclear (RMN) de 1H e 13C. Após isolamento, a substância foi utilizada como padrão para validação de metodologia de quantificação da artepilina C em própolis verde por CLUE-EM-Q-TOF. A metodologia mostrou-se específica, linear, precisa, exata e robusta nas condições avaliadas. Em relação aos ensaios biológicos frente ao nematoide C. elegans, a artepilina C não foi tóxica ao animal, em nenhuma das concentrações avaliadas. Contudo, a artepilina C (125 μM) não foi capaz de prolongar seu tempo de vida. Em embriões de Zebrafish, a substância foi tóxica nas concentrações de 250, 125 e 50 μM, indicando a necessidade de estudos mais aprofundados quanto à sua embriotoxicidade. / Artepillin C is the main chemical marker used in quality control of green propolis, which is produced especially in Minas Gerais and marketed mainly to Japan, being Baccharis dracunculifolia DC. (Asteraceae) its main botanical source. Among the techniques that can be used for the analysis of artepillin C, ultra high performance liquid chromatography (UHPLC) coupled with mass spectrometry (MS) has many advantages. Regarding its biological activities, artepillin C shows antioxidant and antitumor action, and its embryotoxicity and antiaging activity have not been evaluated yet. The aim of this work was to isolate artepillin C, to develop and validate the analytical method of quantification of this substance by UHPLC-MS-Q-TOF, as well as to determine its antiaging activity in Caenorhabditis elegans nematodes and its toxicity in Zebrafish embryos. To isolate the artepillin C from green propolis sample, basic extraction was performed followed by acidification, with subsequent fractionation by vacuum liquid chromatography (VLC) and semipreparative high performance liquid chromatography (HPLC). HPLC and nuclear magnetic resonance (NMR) 1H and 13C analysis were performed for the substance identification. After isolation, the substance was used as standard for validation of quantification methodology of artepillin C in green propolis by UHPLC-MS-Q-TOF. The methodology was specific, precise, accurate and robust for the evaluated conditions. About the biological assays in C. elegans nematode, artepillin C was not toxic for the animal at any of evaluated concentrations. However, artepillin C (125 μM) was not able to prolong its life span. In Zebrafish embryos, the substance was toxic at 250, 125 e 50 μM concentrations, showing the need for further studies about its embryotoxicity.

Orientador: Cláudia Vianna Maurer-Morelli / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-23T06:03:12Z (GMT). No. of bitstreams: 1 Reis-Pinto_FernandaChristina_M.pdf: 1859636 bytes, checksum: b8c3841af89fe165c03148ebcc2437ce (MD5) Previous issue date: 2013 / Resumo: O uso de modelos animais tem trazido grandes benefícios para o conhecimento dos processos biológicos normais, bem como para uma maior compreensão das doenças humanas, inclusive as epilepsias. Embora os modelos de roedores sejam os mais usados para explorar os mecanismos que permeiam a epileptogênese, recentemente, um novo modelo animal foi proposto para o estudo das epilepsias, o Daniorerio. Popularmente conhecido como zebrafish, este pequeno peixe possui muitas vantagens para a experimentação científica, principalmente no que se refere à manipulação genética.O Fator Neurotrófico Derivado do Cérebro (BDNF, do inglês, BrainDerivedNeurotrophicFactor) é a neurotrofina de maior abundância no sistema nervoso central e que está relacionada à plasticidade neuronal. Sabe-se que os níveis de transcritos e protéicos desta neurotrofina estão alterados nas epilepsias, tanto em humanos quanto em modelos animais; porém, o seu papel nesta condição ainda é controverso. Os principais objetivos deste estudo foram (i) investigar o perfil temporal de transcritos dos genes bdnfe seus receptores ntrk2a e ntrk2b após crise epiléptica aguda e (ii) após crises epilépticas repetitivas por meio da técnica da PCR quantitativa (qPCR) usando-se o sistema TaqManTM (AppliedBiosystems, Foster City). As crises epilépticas foram induzidas quimicamente por Pentilenotetrazol (PTZ) em animais adultos e seus encéfalos coletados nos tempos: (i) 0,05h, 12h, 24h, 48h, 72h pós-crise aguda e (ii) uma semana após indução de crise diária por cinco dias consecutivos. Os resultados mostraram que as crises epilépticas modificaram o perfil de expressão das neurotrofinas e apontou para um aumento dos níveis de RNAm do gene bdnf no tempo 0,05h (p= 0,01) que não é descrito em outros modelos. É importante salientar que esses resultados mostram que, no encéfalo do zebrafish, as neurotrofinas estão relacionadas com uma atividade neuronal anormal como em outros modelos animais e em humanos. Este é o primeiro estudo a investigar os níveis de transcritos do gene bdnf e seus receptores no encéfalo do zebrafish, contribuindo para a caracterização deste animal como modelo para estudos das epilepsias / Abstract: Animal models have been contributing to a better understanding of human diseases, including epilepsies. Although rodent models are common organisms used to investigate the mechanisms underlying epileptogenesis, recently, Daniorerio has gained attention as a promising model for epilepsy studies. Popular named as zebrafish, this little fish has many advantages for scientific investigation, especially those related to genetic manipulations. BDNF (Brain Derived Neurotrophic Factor) is the main neurotropic factor in the central nervous system and it is related to neuronal plasticity. It has been reported that both BDNF transcript and protein levels are increased after seizures in patients and experimental models; however, the temporal transcript profile of the BDNF in the zebrafish brain is unknown. The main aim of this study was to investigate the transcripts profile of bdnf, and ntrk2antrk2b genes by quantitative PCR (qPCR) using TaqManTM system (Applied Biosystems, Foster City), (i) after a single seizure and (ii) repetitive seizures. Seizures were chemically induced by Pentylenetetrazole (PTZ) in adult animals and their brains collected at (i) 0,05h, 12h, 24h, 48h, and 72h after acute seizure and (ii) one week after single seizure/day by five consecutive days. Our results showed that in the zebrafish brain, changes in neurotrophins transcript levels were related to an abnormal neuronal activity as seems in other experimental models and patients and presented an increase of bdnf mRNA levels at 0,05h (p=0.01) described for the first time. This is the first study exploring transcript profile of neurotrophins in zebrafish brain and contributes to characterize it as a model for epilepsy studies / Mestrado / Ciencias Biomedicas / Mestra em Ciências Médicas

Depuis le début des années 1990, de nombreuses équipes de recherche ont consacré leurs travaux à étudier les effets des perturbateurs endocriniens (PEs) sur l'homme et les espèces animales. Plusieurs polluants libérés dans l’environnement tels que les pesticides, les herbicides, les plastifiants industriels ou les résidus de médicaments ont ainsi le potentiel de perturber les équilibres hormonaux des organismes, induisant ainsi de graves conséquences sur leur développement et leur reproduction. Pour lutter contre ces dommages, diverses réglementations spécifiques et des systèmes de criblages se mettent en place pour détecter ces molécules. Ces systèmes doivent être rapides et fiables et les poissons téléostéens comme le medaka ou le poisson zèbre constituent d'excellents modèles pour détecter la présence de ces molécules et étudier leurs effets in vivo. Durant cette thèse nous avons étudié l’impact de certains PEs sur le développement embryonnaire en utilisant le poisson zèbre et le médaka comme modèle. Nous avons pu caractériser l'action du Bisphenol A sur la formation des otholites de l’oreille interne du poisson zèbre. En alliant des approches pharmacologiques et génétiques, nous avons identifié le récepteur nucléaire orphelin ERR comme une nouvelle cible in vivo de cette molécule. En parallèle, nous avons développé des lignées transgéniques rapportrices de médaka permettant de détecter la présence de polluants à activités androgénique ou anti-androgénique ce qui accroît ainsi la gamme des outils disponibles pour évaluer la présence des PEs dans les effluents liquides. / Since the early 1990s, many research teams have devoted their work to study the effects of endocrine disruptors (EDCs) in humans and animal species. Many pollutants released into the environment such as pesticides, herbicides, industrial plasticizers or drug residues have the potential to disrupt bodies’ hormonal balances, thus inducing a severe impact on their development and reproduction. To fight against these damages, various specific regulations and screening systems are setting up to detect these molecules. These systems must be fast, reliable. Teleost fish such as medaka and zebrafish are excellent models to detect the presence of these molecules and study their effects in vivo. In this thesis, we studied the impact of certain EDCs on the embryonic development using zebrafish and medaka as models. We characterized the action of Bisphenol A on the otoliths formation in the inner ear of zebrafish. By combining genetic and pharmacological approaches, we identified the orphan nuclear receptor ERRγas a new target of this molecule in vivo. In parallel, we have developed reporter transgenic lines of medaka to detect the presence of pollutants containing androgenic or anti-androgenic activities and thereby increasing the range of tools available to assess the presence of EDCs in liquid effluents.

A biologia molecular e celular tem acumulado um elevado número de fatos experimentais, como seqüências genômicas e propriedades de proteínas. Entretanto, isso não é suficiente para a compreensão dos sistemas biológicos e nem para explicar as intrincadas interações dinâmicas entre a expressão gênica e o desenvolvimento anatômico. É preciso fazer uso de métodos e ferramentas que permitam caracterizar e analisar a complexidade das interações entre o genótipo e o fenótipo, de forma a confrontar as alterações na anatomia com os padrões espaço-temporais da expressão gênica. Neste contexto, o presente trabalho propõe métodos computacionais que, a partir de imagens contendo informações anatômicas e/ou de expressão gênica, em duas e três dimensões, identificam diferentes propriedades geométricas de amostras biológicas, com a finalidade de caracterizar e analisar a relação entre anatomia e expressão gênica. O desenvolvimento desses métodos inclui o uso intensivo de técnicas de processamento de imagens e visualização computacional. Em particular, quatro propriedades geométricas são descritas: o gradiente, a simetria, a distância e a organização espacial. Para ilustrar o potencial dos métodos propostos, são usadas diversas espécies e são analisadas diferentes questões biológicas, aqui listadas: caracterização morfológica e da expressão gênica durante a cardiogênese em embriões de Zebrafish (Danio rerio); caracterização morfológica do desenvolvimento do parasita Echinococcus granulosus; e análise dos danos morfológicos em retinas de traíras (Hoplias malabaricus) causados por intoxicação mercurial. Os resultados obtidos nessas análises permitiram descrever e inferir importantes propriedades do desenvolvimento, da morfologia e dos padrões de expressão gênica desses sistemas biológicos. Com isso, é mostrado que o uso de métodos computacionais capazes de identificar e analisar, a partir de imagens, padrões espaciais de expressão gênica e sua relação com o desenvolvimento morfológico, está se tornando cada vez mais importante para o estudo do desenvolvimento, por fornecerem informações valiosas a respeito da função biológica e das interações entre os genes. / The areas of cell and molecular biology have accumulated a large number of experimental facts, such as genomic sequences and protein structure. However, this is not sufficient to the understanding of biological systems, even to explain the intricate dynamical interactions between gene expression and anatomical development. In order to characterize and analyze the complexity of the interactions between the genotypes and phenotypes, it is necessary to make use of methods and tools that allow the confrontation between spatio-temporal gene expression and changes in developing anatomy. In this context, the present work proposes a set of computational methods that, based on images containing anatomical and/or gene expression information in two and three dimensions, allow the identification of different geometrical properties in biological samples, so as to characterize and analyze the relationship between anatomy and gene expression. The development of these methods includes intensive use of techniques from image processing and computer visualization. In particular, four geometrical properties are described here: gradient, symmetry, distance and spatial organization. The potential of the proposed methods is illustrated in the analysis of various problems, involving different biological issues and species, including: gene expression and morphological characterization of the heart development in Zebrafish (Danio rerio); morphological characterization of the development of the parasite Echinococcus granulosus; and the analysis of the morphological damage in the retina of the fish Hoplias malabaricus caused by mercurial intoxication. The results obtained in these analyses allowed to infer and describe important proprieties with respect to development, morphology and gene expression patterns of these biological systems. Overall, it is shown that the use of computational methods to identify and analyze, from 2D and 3D images, the spatial patterns of gene expression and its relationship with the morphological development, is becoming an increasingly important tool for the study of the development, providing valuable information about the biological function and interactions among genes.

La prise de décision collective est un des mécanismes usités par les espèces sociales lors d’événements de collectes de nourriture, d’optimisation de chemins, de mouvements collectifs, de fuites face à un prédateur ou bien même de sélections d’habitat. Les zebrafish (Danio rerio), animaux grégaires de référence en biologie, n’ont jamais été étudiés dans des contextes de choix collectifs binaires et ils nous semblait légitime d’analyser leurs comportements décisionnels. L’objectif de cette thèse est de répondre aux attentes du projet ASSISI|bf de création d’un système expérimental modulable capable d’accueillir robots comme poissons, de mise au point d’une société mixte adaptative auto-organisée, et de caractérisation des comportements individuels et collectifs des zebrafish implémentables dans les robots. Nos observations montrent que la prise collective de décision et la cohésion des groupes en environnement ouvert, et présentant deux points de repère identiques, sont des processus dynamiques qui varient selon la souche de Danio rerio (AB ou TL) et sont liées à la densité des populations. Constamment en mouvement, les poissons oscillent entre les deux points de repère identiques installés dans leur environnement. La souche AB est toujours plus attirée par ces hétérogénéités et est beaucoup moins cohésive que la souche TL. En environnement contraint, de type deux chambres reliées par un couloir, nous réitérons le même type d’expérience chez la souche la plus à même de prendre des décisions collectivement (AB) et en faisant varier la densité des groupes de 1 à 20 individus. Il ressort que les Danio rerio de type AB restent essentiellement en groupe mais que l’augmentation de leur densité tend à les diviser. Nous remarquons aussi que la densité influe partiellement sur la topologie du groupe : il existe, lors des sorties collectives, une corrélation entre l’ordre de sortie des zebrafish (les suiveurs) et l’ordre de distance de chaque individu par rapport à l’initiateur desdites sorties. Cette corrélation est d’autant plus forte quand les suiveurs sont soit très proches, soit les plus éloignés de l’initiateur. Le dispositif expérimental précédent nous permet d’autre part de nous concentrer sur la notion de leadership chez des groupes de 2 à 10 Danio rerio AB. Nous mettons en évidence que chaque poisson initie au moins une fois un départ collectif, le nombre d’initiations effectives est proportionnel au nombre de tentatives d’initiations, que tous les poissons présentent le même taux de succès d’initiation après une tentative et qu’il existe une corrélation positive entre initiations et vitesse moyenne de nage.Une analyse poussée des mouvements collectifs nous fait constater que les zebrafish se déplacent essentiellement en groupe et transitent sans interruption ni périodicité entre les chambres. Nous sommes actuellement en train d’améliorer cette étude en apportant de nouveaux points de comparaison afin de vérifier si la non-périodicité des transitions entre les chambres est liée à la densité des groupes testés. Enfin, nous nous inspirons du précédent système expérimental – les deux chambres reliées par un couloir – pour créer une version actualisée d’un labyrinthe en Y. Ce labyrinthe en Y est dit perpétuel car il combine tout le savoir faire que nous avons développé dans l’élaboration des précédents systèmes expérimentaux : une faible habituation des zebrafish pour leur milieu, une réduction des interactions entre l’expérimentateur et les animaux et la possibilité pour les poissons de circuler dans un labyrinthe à symétrie centrale, sans zone de départ ni zone de fin formalisées (typiques des labyrinthes en Y) / The collective decision-making is one of the mecanisms used by social species during foraging, path optimisation, collective movements, predator avoidance or habitat selection. Zebrafish (Danio rerio), gregarious animal models in biology, have never been studied in the context of collective binary choices. We found logical to analyse their decision behaviours. The goal of this thesis is to meet the expectations of the ASSISI|bf project of creation of an andjustable experimental setup able to sustain robots and fish, of development of an adaptive and self-organised mixed society and of the caracterisation of individual and collective behaviours of the zebrafish, implementable in the robots. We show that the collective decision making and the cohesion of groups of zebrafish in an open environment, composed of two identical landmarks, are dynamical processes that vary in function of the strain of Danio rerio (AB or TL) and are related to the density of the populations. Always on the move, the fish oscillate between the two landmarks. The AB zebrafish are more attracted by the landmarks and are less cohesive than the TL zebrafish. In a contrained environment, two rooms connected by a corridor, we do the same type of experiments with only AB zebrafish and vary the density of the groups from 1 to 20 individuals. AB zebrafish swim together. The increase of the density of the groups make them split. We noticed that the density of the groups has an effect on the topology of the groups : during collective departures from a room to the other one, there is a correlation between the rank of exit of the zebrafish (followers) and the rank of the distances of each individual from the initiator of the exits. This correlation is higher when the followers are either the closest or the farest from the initiator.Using the previous experimental setup, we focused also on the topics related to leadership for groups of 2 to 10 individuals. We show that each fish intiate at least once a collective departure and that the number of initiations is proportional to the number of initiation attempts. Also, we found that all the fish have the same success rate in the initiations after an initiation attempt and that there is a positive correlation between the intiations and the average speed. A deeper analysis of the collective movements shows that the zebrafish swim in group and transit without any interruption or even without periodicity between the rooms. We are currently improving this study and will bring new points of comparison to check if the non-periodicity of the transitions could be related to the density of the groups. Finally, we developed a new version of the Y-maze based on the previous observations.This perpetual Y-maze shows many usefull features for the study of the behaviour : a low habituation rate, a reduction of the interactions between the animals and the experimentators, and allows the fish to move in a Y-maze without starting nor ending zones as in typical Y-maze