Дисертації з теми "Drosophila melanogaster dopamine transporter"
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Piazzesi, Antonia Felicia. "Drosophila melanogaster as a model for mitochondrial biology, mitochondrial disease and neurological disorders." Doctoral thesis, Università degli studi di Padova, 2014. http://hdl.handle.net/11577/3423704.
Повний текст джерелаDrosophila melanogaster ha una lunga storia come animale modello per tante malattie umane, incluse le malattie genetiche, la neurodegenerazione e l’alcolismo. Anche se Drosophila fu inizialmente utilizzata come animale modello più di 100 anni fa, rimane comunque un modello rilevante oggi grazie al suo ciclo vitale breve, il suo basso costo e la sofisticazione degli attrezzi molecolari disponibili per la sua manipolazione genetica. Questo modello ha anche meno ridondanza genetica rispetto ai mammiferi, rendendo lo studio della funzione di questi geni molto più diretto, ma malgrado questo possiede un ortologo per 75% dei geni legati a malattie umane. Tutte queste proprietà contribuiscono alla sua rilevanza come modello e sono state sfruttate durante questo progetto. In primis, Drosophila melanogaster è stata usata come modello per il trasporto mitocondriale di deossinucleotidi. Il gene RIM2 in lievito, che è stato precedentemente caratterizzato come trasportatore mitocondriale di deossinucleotidi pirimidinici, ha un omologo in Drosophila: CG18317, qui chiamato drim2, che è stato caratterizzato in questo progetto. Questo gene è stato rimosso in vivo e la funzione mitocondriale e l’integrità del mtDNA sono state caratterizzate. I due omologhi umani per questo gene, PNC-1 e SLC25A36, sono stati espressi nel mutante, per determinare l’omologia funzionale di questi geni e per confermare la validità di questo mutante come modello per il trasporto mitocondriale umano di deossinucleotidi. Questo progetto si è anche focalizzato su una caratterizzazione più approfondita di una linea mutante per dTTC19, un omologo di un gene umano che è già stato collegato alla encefalopatia mitocondriale e la psicosi. Questa caratterizzazione è stata accompagnata dalla generazione di tre linee mutanti che esprimono dTTC19, per confermare che il fenotipo mutante osservato sia dovuto alla mancanta trascrizione di dTTC19. In fine, questo progetto propone un nuovo protocollo che, nella nostra opinione, permetterà di utilizzare Drosophila melanogaster come modello per disturbi neurologici che presentano con sintomi asociali. Un protocollo è stato sviluppato per studiare il comportamento sociale in Drosophila melanogaster e per dimostrare che piccole differenze nei livelli di dopamina o nel contatto sociale dopo l’eclosione possono avere effetti drammatici sulle interazioni sociali in Drosophila. Proponiamo che Drosophila può essere un modello utile per lo studio dei fattori genici coinvolti nelle malattie che presentano con comportamento asociale come l’autismo, il disturbo ossessivo compulsivo, la depressione eccetera. In conclusione, questo progetto sfrutta interamente Drosophila melanogaster come animale modello per la biologia e le malattie mitocondriali. In più, propone un nuovo modo per utilizzare Drosophila come modello che non è stato finora sfruttato.
Gmeiner, Florian [Verfasser], and Charlotte [Gutachter] Förster. "Der Einfluss der Neurotransmitter Dopamin, Serotonin und GABA sowie ihrer Transporter auf das Schlafverhalten von Drosophila melanogaster / Florian Gmeiner. Gutachter: Charlotte Förster." Würzburg : Universität Würzburg, 2014. http://d-nb.info/1108780822/34.
Повний текст джерелаMeyer, Heiko. "Ein neuer Zuckertransporter in Drosophila melanogaster." Doctoral thesis, [S.l.] : [s.n.], 2006. http://deposit.ddb.de/cgi-bin/dokserv?idn=979030331.
Повний текст джерелаJohnson, Kevin. "Isolation and characterisation of a novel neurotransmitter transporter in Drosophila melanogaster." Thesis, University of Cambridge, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.387584.
Повний текст джерелаEvans, Jennifer Mary. "From microarrays to renal transporter function in Drosophila melanogaster Malpighian tubules." Thesis, University of Glasgow, 2007. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.443441.
Повний текст джерелаMarican, Charlotte. "Dissection génétique du rôle des catécholamines dans le comportement sexuel de Drosophila melanogaster." Paris 11, 2002. http://www.theses.fr/2002PA112314.
Повний текст джерелаSexual behavior in Drosophila is genetically determined. It implies different communication mechanisms (chemical : pheromones; acoustic : song and visual) and simpler behaviors ( like locomotor activity). In addition, it has been demonstrated that terminal enzymes for dopamine synthesis (DDC : dopa decarboxylase and TH : thyrosine hydroxylase) were formed under the cuticle and in the central nervous system. Initially, our research focused on potential differences in cuticular hydrocarbons, linked to the presence or not of dopamine. For that, genetical and pharmalogical approaches were used: TH enzyme inhibitor on wild type strains, mutant strains (Ddc ts) complemented with their lacking compounds and mutant strains for their structural genes: Ddc and pale. We observed a correlation between diminution of dopamine and of female-specific hydrocarbons. Then, we demonstrated that this defect, comparable to the decapitation effect, is compensated for the females by a single topical application of dopamine. We also analysed different behavioral aspects. We demonstrated that male flies having reduced levels of dopamine and serotonine have a lower success rate for copulation, a lower locomotor activity and a modified song. Dopamine seems to play a significant role in sexual behavior regulation at different levels : pheromons production, acoustic production, and locomotor activity. Finally, a study of pheromonal variations as a function of temperature bas been performed. It shows that higher temperatures favor the production of longer chains of hydrocarbons either in males and females in two wild type strains of D. Melanogaster
Frighetto, Giovanni. "Selection for action in Drosophila melanogaster." Doctoral thesis, Università degli studi di Padova, 2019. http://hdl.handle.net/11577/3424669.
Повний текст джерелаIl presente lavoro si focalizza sui meccanismi di selezione per il controllo dell’azione utilizzati da Drosophila melanogaster (D. melanogaster), anche nota come moscerino della frutta. D. melanogaster ha un ricco repertorio di comportamenti e un cervello semplice, composto di circa 100,000 neuroni, che può essere studiato con tecniche raffinate. Perciò, offre la possibilità di studiare un comportamento in una struttura cerebrale semplice rispetto a quella di organismi più complessi. Le basi neurobiologiche del suo comportamento possono essere così più facilmente comprese. Il confronto con comportamenti simili mostrarti da animali evolutivamente più lontani può fornire importanti intuizioni sui circuiti neurali sottesi. Questa tesi è stata concettualizzata per verificare se i processi di selezione per l’azione dei mammiferi potessero essere condivisi con organismi più bassi come D. melanogaster. La selezione per l’azione implica una stretta interazione tra sistema visivo e motorio che consente di scegliere uno stimolo nell’ambiente per agire su di esso. Questo processo permette di filtrare le informazioni irrilevanti per l’azione. Il primo esperimento era finalizzato a indagare se i moscerini mostrano un’attenzione basata sull’azione. Sono capaci di inibire la risposta a uno stimolo grazie a meccanismi attentivi per terminare un’azione? In particolare ho indagato se i moscerini sono inclini all’effetto d’interferenza causato dalla comparsa di uno stimolo competitivo (cioè, un distrattore). Mi aspettavo che questo meccanismo inibitorio fosse evidente nelle traiettorie spaziali. In questo studio i moscerini erano impegnati a raggiungere un target visivo (cioè, una striscia luminosa) mentre un distrattore compariva lateralmente. Il secondo esperimento mirava a estendere le conclusioni del primo esperimento. In particolare sono state considerate le distanze tra target e distrattore. L’obiettivo di questo studio era di verificare l’ipotesi che più corta è la distanza tra target e distrattore, più elevato è il livello d’inibizione. Nel terzo esperimento ho individuato l’ipotetico circuito neurale responsabile degli effetti comportamentali osservati negli esperimenti precedenti. Basandomi su crescenti evidenze in favore di un’affascinante omologia tra un neuropilo dei moscerini (il Complesso Centrale, CC) e la struttura neurale dei mammiferi implicata nella selezione dell’azione, ho testato il comportamento in moscerini con CC danneggiato. A questo scopo ho usato una tecnica basata sul sistema binario GAL4-UAS per ridurre l’espressione di specifici recettori dopaminergici in un circuito neurale molto selettivo, i cosiddetti neuroni E-PG. Inoltre, ho adottato una tecnica optogenetica per la manipolazione neurale in vivo. Ho utilizzato moscerini che esprimevano canali ionici foto-attivabili nello stesso circuito del CC per eccitare tali neuroni durante il compito. Questo circuito neurale forma una struttura a ciambella che è stata considerata un centro d’integrazione tra il sistema visivo e motorio deputato all’attenzione. Infine, nel quarto esperimento, ho caratterizzato da un punto di vista neurochimico una serie di circuiti neurali del CC. L’ipotesi era che il sistema dopaminergico implicato nel processo di selezione dell’azione nei mammiferi potesse modulare anche nei moscerini la risposta neurofisiologica all’interno del CC. Nello specifico, ho registrato in vivo la risposta neurale ad applicazioni di dopamina nel CC dei moscerini utilizzando una tecnica di bioluminescenza basata su indicatori del calcio geneticamente codificati. Nel complesso questo lavoro rappresenta un tentativo di affrontare i meccanismi di selezione per il controllo dell’azione nei moscerini. Il paradigma d’interferenza che ho sviluppato costituisce una potente piattaforma per esplorare il problema della selezione per l’azione che potrebbe essere utile al fine di chiarire processi simili in organismi più complessi.
Trannoy, Séverine. "Analyse fonctionnelle de circuits neuronaux impliqués dans la dynamique des mémoires olfactives chez Drosophila melanogaster." Paris 6, 2012. http://www.theses.fr/2012PA066337.
Повний текст джерелаWhen we present an odor associated with sugar to starved flies, they will be attracted by this odor. One cycle of conditioning induces both Short-Term Memory (STM) and Long-Term Memory (LTM). It is accepted that STM and LTM formation is a sequential process but the link between these two memories remains unknown. We adressed this question and clearly demonstrated that STM and LTM can be formed independently and that they involved different neural structures within the Mushroom Bodies (MB), a memory center. We proposed a new model of dynamic of appetitive memory phases where STM and LTM are formed in a parallel way. Then, using the genetically expressed thermosensible toxine allowing a transiently inactivation of neurotransmission, we identified one type of MB efferent neurons involved in appetitive LTM retrieval. Additionally, we were interested to the molecular mechanisms and the neuronal circuits involved in aversive consolidated memories. Pairing an odor with electric shocs induces aversive memory. In drosophila, there are two forms of consolidated memories, the Anesthesia-Resistant Memory (ARM) and LTM (dependent on de novo protein synthesis). We show that three pairs of oscillatory dopaminergic neurons play a essential role of gating between ARM and LTM formation. So, blocking the neurotransmission of these neurons during the consolidation phase leads to a increase of ARM and inhibition of LTM whereas, artificial activation of these neurons after conditioning leads to an inhibition of ARM and favors the implementation LTM. In conclusion, we characterized functionally a restricted population of neurons playing a role in various stage of learning and memory process
Meehan, Michael J. "A behavioural and pharmacological study of mutants that influence amine metabolism in Drosophila melanogaster." Thesis, University of Ulster, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.292841.
Повний текст джерелаCruz, Melissa Joy. "Functional characterization of heterologously expressed Drosophila melanogaster organic cation transporter ORCT in Xenopus laevis oocytes." Thesis, University of British Columbia, 2015. http://hdl.handle.net/2429/52554.
Повний текст джерелаIrving K. Barber School of Arts and Sciences (Okanagan)
Biology, Department of (Okanagan)
Graduate
Fagan, Rita R. "Rit2-Dependent Dopamine Transporter Endocytosis: Intrinsic Mechanism and In Vivo Impact." eScholarship@UMMS, 2020. https://escholarship.umassmed.edu/gsbs_diss/1086.
Повний текст джерелаAssael-Monier, Magdalena. "Mate copying chez la drosophile : importance évolutive et bases mécanistiques." Thesis, Toulouse 3, 2020. http://www.theses.fr/2020TOU30040.
Повний текст джерелаMate-copying has been reported in many Vertebrate and Invertebrate species, including animals as simple in appearance as Drosophila melanogaster. In this species, when a female observes another female mating with a male of a given phenotype, his attraction to other males of this phenotype increases. In other words, she copies the mate preference of the demonstrator female. This behavior constitutes a powerful model of social observational learning in animals, both for proximate mechanisms (for instance behavioral and neurobiological) as well as ultimate mechanisms (notably, as it takes part to sexual evolution). The present work studied these two aspects of mate-copying. The first chapter tested the stability of mate-copying across environmental social conditions, more specifically, apparent availability of males, and across time (long-term memory). I showed that, while sex-ratio affects female choosiness positively, Drosophila females seem to have evolved a mate-copying ability independently of sex-ratio. I also participated in showing that females can form a social long-term memory (24h) involving protein synthesis. Chapters 2 and 3 deal with cognitive mechanisms in mate-copying. I showed that it involves the neurotransmitters dopamine and serotonine, while the dopaminergic receptor DAMB (DopAmine Mushroom Bodies) is required for this social long-term memory, but not for short-term memory, which suggests that another dopaminergic receptor is also involved in this social learning. I designed and tested a new protocol of demonstrations based on photographs, which will ease the study of the visual cues necessary for this behavior, and later the study of the neurobiological mechanisms. Finally, I showed that mate-copying is a learning based on on the trait of the male accepted by the demonstrator female, and not on the rejected one, and I found that, counter-intuitively, dopaminergic networks involved are those for aversive, not appetitive, olfactory learning
Burke, Christopher J. "Layered Reward Signalling Through Octopamine and Dopamine in Drosophila: A Dissertation." eScholarship@UMMS, 2013. http://escholarship.umassmed.edu/gsbs_diss/657.
Повний текст джерелаOhmann, Elisabeth [Verfasser], and Stephan [Akademischer Betreuer] Schneuwly. "Untersuchung zur Funktion und Bedeutung der ABC-Transporter in Drosophila melanogaster / Elisabeth Ohmann. Betreuer: Stephan Schneuwly." Regensburg : Universitätsbibliothek Regensburg, 2011. http://d-nb.info/1023281961/34.
Повний текст джерелаTitlow, Josh S. "Dopaminergic and Activity-Dependent Modulation of Mechanosensory Responses in Drosophila Melanogaster Larvae." UKnowledge, 2014. http://uknowledge.uky.edu/biology_etds/21.
Повний текст джерелаAraujo, Stífani Machado. "Efeito protetor do γ-orizanol em um modelo de doença de parkinson induzida por rotenona em Drosophila melanogaster". Universidade Federal do Pampa, 2016. http://dspace.unipampa.edu.br:8080/xmlui/handle/riu/313.
Повний текст джерелаMade available in DSpace on 2016-04-18T18:12:36Z (GMT). No. of bitstreams: 1 Stífani Machado Araujo.pdf: 2701104 bytes, checksum: db0f252b8e5b648fd9a2f147be16081a (MD5) Previous issue date: 2016-02-19
Doença de Parkinson (DP) é a segunda doença neurodegenerativa mais comum no mundo, afetando cerca de 1% dos adultos com mais de 60 anos. A DP está relacionada com a degeneração de neurônios dopaminérgicos principais componentes da substância negra cerebral, concomitantemente, com a disfunção do complexo I mitocondrial e o estresse oxidativo que desempenham um papel crucial na patogênese desta doença. A rotenona (ROT) é um pesticida natural e muito utilizado para induzir fenótipo de DP em modelos animais, por ser lipofílico pode atravessar facilmente a barreira hematoencefálica causando disfunção do complexo I mitocondrial e possível morte de neurônios dopaminérgicos. Dentre as várias aplicações terapêuticas dos antioxidantes, ressalta-se sua ação neuroprotetora, uma vez que o sistema nervoso central exibe uma maior vulnerabilidade e susceptibilidade aos insultos oxidativos, o γ-orizanol (ORY) é um produto natural composto por uma mistura de ésteres de ácido ferúlico extraídos a partir do óleo de farelo de arroz, e é bem descrito na literatura por possuir propriedades antioxidantes. Assim, o objetivo deste trabalho foi investigar um possível efeito neuroprotetor do γ-orizanol sobre alterações comportamentais e bioquímicas causadas pela exposição crônica de Drosophila melanogaster a rotenona. A mosca da fruta Drosophila melanogaster, é uma espécie alternativa à utilização de modelos mamíferos que vem sendo usada com bastante confiabilidade na reprodução de modelos de disfunção dopaminérgica. As moscas (de ambos os sexos) com idades compreendidas entre 1 a 5 dias de idade foram divididos em quatro grupos de 50 moscas cada um: (1) de controle, (2) ORY 25 μM, (3) ROT 500 μM, (4) ORY 25 μM + ROT 500 μM. As moscas foram concomitantemente expostos a uma dieta contendo ROT e ORY durante 7 dias de acordo com os seus respectivos grupos. Após o tratamento foram feitas as analises comportamentais e bioquimicas. Como resultados, verificamos que o ORY ofereceu proteção contra as alterações locomotoras causadas por ROT, além de prevenir a mortalidade induzida por rotenona, protegeu contra geração de estresse oxidativo e disfunções mitocondriais além de otimizar as defesas antioxidantes celulares. Nossas descobertas apontam uma restauração dos déficits colinérgicos, e nos níveis de dopamina fornecida por pelo tratamento com ORY. Em conclusão, os presentes resultados mostram que ORY é eficaz na redução da toxicidade induzida ROT em Drosophila melanogaster, o que mostrou uma ação neuroprotetora, possivelmente devido à presença dos componentes de antioxidantes tais como o ácido ferúlico.
Parkinson's disease (PD) is the second most common neurodegenerative disease in the world, affecting about 1% of adults over 60 years. The DP is linked to the degeneration of dopaminergic neurons main components of the substantia nigra brain, concurrently with mitochondrial complex I dysfunction and oxidative stress play a crucial role in the pathogenesis of this disease. The rotenone (ROT) is a natural pesticide, and used to induce PD phenotype in animal models, being lipophilic can easily cross the blood-brain barrier dysfunction causing mitochondrial complex I and possible death of dopaminergic neurons. Among the various therapeutic applications of antioxidants, it emphasizes its neuroprotective action, as the central nervous system displays a greater vulnerability and susceptibility to oxidative insults, the γ-oryzanol (ORY) is a natural product composed of a mixture of esters ferulic acid derived from rice bran oil, and is well described in the literature to possess antioxidant properties. The objective of this study was to investigate a possible neuroprotective effect of ORY on behavioral and biochemical changes caused by chronic exposure of Drosophila melanogaster the rotenone. The fly fruit Drosophila melanogaster, is an alternative species to the use of mammalian models that have been used quite reliable reproduction of dopaminergic dysfunction models. The flies (male and female) between the ages of 1 to 5 days of age were divided into four groups of 50 flies each: (1) control, (2) ORY 25 μM, (3) ROT 500 μM ( 4) ORY 25 μM + ROT 500 μM. The flies were simultaneously exposed to a diet containing ROT and ORY for 7 days according to their respective groups. After treatment were made behavioral and biochemical analysis. As a result, we find that the ORY offered protection against locomotor changes caused by ROT, and to prevent the mortality induced by rotenone, protected against the generation of oxidative stress and mitochondrial dysfunction and optimize cellular antioxidant defenses. Our findings point to a restoration of cholinergic deficits, and dopamine levels provided by the treatment ORY. In conclusion, the present results show that ORY is effective in reducing the ROT induced toxicity in Drosophila melanogaster, which showed a neuroprotective effect, possibly due to the presence of antioxidants components such as ferulic acid.
Friggi-Grelin, Florence. "Régulation génétique de la différenciation des cellules dopaminergiques : contrôle transcriptionnel et post-transcriptionnel de l'expression de la tyrosine chez Drosophila melanogaster." Aix-Marseille 2, 2001. http://www.theses.fr/2001AIX22066.
Повний текст джерелаSocha, Catherine. "Study of the metabolic aspects of resilience to intestinal infections in Drosophila melanogaster." Thesis, Strasbourg, 2018. http://www.theses.fr/2018STRAJ106.
Повний текст джерелаUpon microbial infections, host defenses comprise two complementary facets. First, immune effectors target and kill the invading pathogen, an attack referred to as resistance. Second, the infected host must repair the damages inflicted by microbes or by the immune response itself, a mechanism called resilience. I have studied the effects of an intestinal infection with the bacterium Serratia marcescens in Drosophila. We have discovered a purge mechanism in the intestine, where enterocytes -the main cell type in the gut- extrude some of their internal contents. The intestinal epithelium thus becomes very thin but rapidly recovers its shape, thereby protecting the fly against the deleterious effects of infection. I have identified an amino acid transporter, CG1139, which is required for the intestinal recovery. CG1139 is necessary to mobilize the fly’s internal metabolic reserves and to transport some these metabolic stores back to the gut, in a retrograde manner
Simonnet, Mégane. "Rôle du transporteur d'acides aminés Minidiscs dans le fonctionnement du système nerveux chez Drosophila melanogaster." Thesis, Dijon, 2015. http://www.theses.fr/2015DIJOS046/document.
Повний текст джерелаAmino acids have many functions in the body in addition to their role as basic constituents of proteins. They can for example serve as a neurotransmitter or signal for the activation of intracellular pathways. Carriers of the SLC protein family facilitate their path through the plasma membrane. The heterodimeric amino acid transporters HAT belong to SLC proteins. HAT are composed of a light chain SLC7 ensuring the specificity of transport and a heavy chain SLC3 involved in the addressing of the protein complex to the plasma membrane. My thesis focused on studying the role of a SLC7 homologue in drosophila, Minidiscs (Mnd), in the functioning of the nervous system. Mnd might belong to system L carriers, mainly known for their role in cell proliferation. My thesis work led to highlight the location of Mnd in the drosophila brain in some neurons (mushroom bodies, dopaminergic neurons) and some glial cells (cortical glia). The presence of Mnd in the brain seems to be involved in the modulation of some behaviors such as negative geotaxis reflex. This work also showed that, as for mammal HAT, Mnd is associated covalently to a protein partner. Transport experiments seem also to confirm the belonging of Mnd to the system L. These results suggest that Mnd is probably involved in the regulation of neuronal activity and thus in the functioning of the nervous system, which had never been described for a system L carrier
Furrer, Marie-Pierre. "Fonction des gènes de la famille elav/Hu dans la différenciation du système nerveux chez Xenopus laevis et Drosophila melanogaster." Paris 11, 2000. http://www.theses.fr/2000PA112370.
Повний текст джерелаThe goal of this thesis was to understand the role of ELAV/Hu family proteins in the development, maintenance and function of neurons, using phenotypic analyses of deregulation mutants in two models. In Xenopus, we showed that 3 elav/Hu family homologs (elrB, elrC, and elrD) are differentially expressed during nervous system development, suggesting a role in different phases of neuron maturation and the definition of different neural sub-domains. Deregulating elrB expression in Xenopus embryos reduced early neuronal markers expression, stopped cell proliferation, and induced apoptosis, suggesting a role in post-transcriptional regulation of genes controlling cell cycle and/or apoptosis. In Drosophila, ubiquitous expression of elav, obtained with the UAS/Gal4 system, produced a phenotype of cuticle and bristles depigmentation. This phenotype, similar to that of mutants of dopamine (DA; neuromodulator and cuticle biosynthesis intermediate in Drosophila) biosynthesis. This depigmentation phenotype suggests that elav could regulate production of neural tyrosine hydroxylase (TH), which control the first step of DA biosynthesis. This work suggests that ELAV/Hu proteins participate in the control of cell cycle and/or apoptosis in addition to their role in neuron differentiation. Identification of TH as a potential target of ELAV suggests a role in controlling the biosynthesis of DA, thus establishing a link between ELAV presence and its function in the nervous system
Coban-Poppinga, Büsra [Verfasser], André [Akademischer Betreuer] Fiala, André [Gutachter] Fiala, and Martin [Gutachter] Göpfert. "Plasticity of Dopamine-Releasing Central Brain Neurons Underlying Adaptational Feeding-Related Behavior in Drosophila Melanogaster / Büsra Coban-Poppinga ; Gutachter: André Fiala, Martin Göpfert ; Betreuer: André Fiala." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2020. http://d-nb.info/1209738546/34.
Повний текст джерелаGmeiner, Florian. "Der Einfluss der Neurotransmitter Dopamin, Serotonin und GABA sowie ihrer Transporter auf das Schlafverhalten von Drosophila melanogaster." Doctoral thesis, 2014. https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-99152.
Повний текст джерелаThe main focus in the present work, was the observation of the influence of dopamine, serotonin and GABA on the sleep behaviour of Drosophila melanogaster. By utilizing mutants for the dopamine transporter as well as the serotonin transporter, it was possible to show, that dopamine and serotonin have opposing effects on the total sleep amount of flies. Dopamine has a sleep inhibiting, serotonin a sleep promoting function. A neuronal dopamine deficient stock complemented those findings. Usage of RNAi to downregulate dopamine receptors did not enhance the information, since no measurable effect could be detected. But in parallel performed experiments with RNAi mediated knockdown of GABABR2 receptors could show its role in the maintenance of sleep during the second half of the night. I could show that especially the expression in the l-LNv is needed for that. In case of the GABABR2 receptors no effect on the period was observed, for dopamine and serotonin only a minor effect on the clock in form of a mild period change accompanied those drastic sleep phenotypes. Combining the amine transporter mutants with functional as well as mutated white led to some interesting observations regarding the interaction of those transporters in regulating total sleep, in which white reduces the total sleep amount. Following up those experiments with UPLC measurements, it was shown that presumably WHITE causes its effect due to its relevance for the amount of beta-alanyldopamine in adult flies. When dopamine is transported into the glia cells, beta-alanyldopamine is synthesized by the enzyme EBONY. The ebony mutant revealed a drastic sleep phenotype when combined with an intact WHITE transporter and a mutated dopamine transporter. This leads to a dramatic decrease of sleep during the night phase. When doing the UPLC measurements it was furthermore revealed, that unexpectedly regarding the knowledge from cell culture experiments, beta-alanylserotonin cannot be detected. Presumably, only dopamine, but not serotonin is recycled by the glia cells. This interesting difference gives space for a temporal as well as for a local fine regulation of the dopamine and serotonin signals. Investigating the dimer partners of WHITE, BROWN and SCARLET, I found that BROWN just as a spontaneous white mutation that I observed, led to a decrease of total sleep. The function of this heterodimer and its neuronal localisation in the brain remains unknown. Regarding the data presented in this work, it is likely that this dimer transports either dopamine or beta-alanyldopamine in glia cells. Furthermore, I could observe that dopamine and serotonin change the ability of the circadian clock to adapt to different photoperiods, a so far unstudied phenotype. 96 An anatomical approach to localize the WHITE transporter in the brain of Drosophila melanogaster and a further characterization of the function of the WHITE/BROWN dimer, with regard to sleep and eventually the mapping of serotonergic and dopaminergic neurons, which modulate the activity peak responses, are questions for future work
Cheng, Kuo-Chang, and 鄭國璋. "Structural and functional studies of dopamine N-acetyltransferase from Drosophila melanogaster." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/12521365222471762062.
Повний текст джерела國立清華大學
生物資訊與結構生物研究所
101
Melatonin is a major hormonal mediator of light-induced photoperiodic changes in circadian biological events and is found in bacteria, protozoa, macroalgae, plants, fungi, invertebrates, and vertebrates. Despite of the structural characterization of melatonin, there has been increasing interest in this “molecular pacemaker” hormone. The physiological roles of melatonin have been widely reported on sleep, mood, immune response, cardiovascular fitness and aging. Research on melatonin biosynthesis could help improve our knowledge of circadian rhythm. The daily cycle of melatonin biosynthesis in mammals is regulated by AANAT (arylalkylamine N-acetyltransferase; EC 2.3.1.87), making it an attractive target for therapeutic control of abnormal melatonin production in mood and sleep disorders. Drosophila melanogaster Dat (dopamine N-acetyltransferase) is an arylalkylamine N-acetyltransferase, which is involved in melatonin formation, sclerotization, and neurotransmitter inactivation and has been found in the head, the eyes, the optic lobe and the brain of Drosophila melanogaster. Moreover, Dat belongs to the GCN5-related N-acetyltransferase (GNAT) superfamily. Dat catalyzes the transfer of the acetyl group in acetyl coenzyme A (AcCoA, cofactor) to various arylalkylamines (substrate). In order to unravel the detail molecular mechanism for Dat activity, we worked on the structural and functional studies of Dat. We have determined high-resolution crystal structure of D. melanogaster Dat in apo form, binary complex form (AcCoA-bound), and ternary complex form (acetylarylalkylamine/CoA-bound). A binding study using isothermal titration calorimetry suggested that the cofactor bound to Dat first before substrate. Examination of the binary complex structure and a substrate-docked model indicated that Dat contains a novel AANAT catalytic triad. Site-directed mutagenesis, kinetic studies and pH-rate profiles confirmed that Glu47, Ser182 and Ser186 were critical for catalysis. Collectively, the results of the present study suggest that Dat possesses a specialized active site structure dedicated to a catalytic mechanism where nucleophilic attack and leaving group protonation occur in a coordinated manner dependent on catalytic triad. The molecular basis of substrate recognition and the kinetic mechanism by which Dat interacts with substrate and cofactor are unclear. Here, two-substrate kinetic analysis and dead end analog inhibition studies with the tryptophol and palmitoyl CoA indicated that Dat utilizes an ordered sequential mechanism requiring binding of acetyl-CoA first. Furthermore, we presented the first crystal structure of ternary complex in this AANAT family. Detailed analyses of ternary complexes of Dat revealed a hydrophobic substrate-binding pocket near the acetylation active site. The shape and size of the pocket dictate substrate selectivity and specificity. We have mapped two key aromatic residues in the protein-substrate interface essential for substrate binding and selection between phenylalkylamines (PAAs), indoalkylamines (IAAs), and arylamines substrates. The Dat has higher activity with the PAAs than with the IAAs. It appears that the aromatic ring and alkyl chain length on arylalkylamine molecule greatly define the Dat extended substrate specificity profile. By analyzing ternary complex structure as well as site-directed mutagenesis, we demonstrated that Phe43 significantly influence the substrate binding and the activity of Dat, while Tyr64 was an important determinant of substrate preference. Kinetic studies confirmed that the Y64W mutation is sufficient to increase the activity of the enzyme toward IAAs being the preferred substrate for the Y64W mutant, which indicated that this residue modulates the substrate preference of Dat between PAAs and IAAs. These results confirmed that these residues are critical for aromatic interaction between Dat and substrate. This work provides a structural foundation for the detailed understanding of the structural and biological properties of arylalkylamine N-acetyltransferases and of GCN5-related N-acetyltransferase superfamily proteins in general.
Lin, Fan-Wei, and 林凡惟. "Study of stustrate-entrance tunnel of Dopamine N-acetyltransferase from Drosophila melanogaster." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/57376575961073600641.
Повний текст джерела國立清華大學
生物資訊與結構生物研究所
102
Drosophila melanogaster dopamine N-acetyltransferase (Dat, EC 2.3.1.87) belongs to the arylalkylamine N-acetyltransferase (AANAT) family, which catalyzes the synthesis of the hormone precursor (melatonin). We have solved the structures of Dat in apo form, binary complex (Dat / acetyl coenzyme A) and ternary complex form (Dat / acetylarylalkylamine / CoA) and proposed the catalytic mechanism previously. According to the binding study by isothermal titration calorimetry (ITC), the cofactor (Acetyl-CoA) needed to bind to the Dat prior to substrate, which would hinder the substrate entry to its binding site. Therefore, we speculate that an entry tunnel for substrate may exist to facilitate the substrate binding to the active site. In this study, we replaced two residues with tryptophan, M121 and D142, located inside the tunnel to see the effects of tunnel hindrance. Our DTNB-based enzyme activity measurements and enzyme kinetic studies showed that mutant M121W decreased the enzyme activity and the substrate binding comparing to wild type Dat. Among the four substrates (Dopamine, serotonin, phenylethylamine, tryptamine) tested, only the efficiency of dopamine remains. This result confirms that M121W and D142W may hinder the substrate entry, resulting in decreased binding efficiency of the binary complex. Our studies not only confirm the existence of a substrate tunnel, but also show the tunnel size may contribute to the substrate specificity.
Kim, Youngcho Han Kyung-An. "Distinctive roles of dopamine and octopamine receptors in olfactory learning of Drosophila melanogaster." 2007. http://etda.libraries.psu.edu/theses/approved/WorldWideIndex/ETD-2095/index.html.
Повний текст джерелаSoehnge, Holly Osborne. "Cloning and sequencing of inebriated, a gene encoding a neurotransmitter transporter in Drosophila melanogaster." Thesis, 1997. http://hdl.handle.net/1911/19214.
Повний текст джерелаMachado, Jerry. "A comprehensive genomic analysis of nucleoside transporters and the functional characterization of the Drosophila equilibrative nucleoside transporter Isoform DmENT2 /." 2004. http://gateway.proquest.com/openurl?url%5Fver=Z39.88-2004&res%5Fdat=xri:pqdiss&rft%5Fval%5Ffmt=info:ofi/fmt:kev:mtx:dissertation&rft%5Fdat=xri:pqdiss:MQ99354.
Повний текст джерелаTypescript. Includes bibliographical references (leaves 46-56). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL: http://gateway.proquest.com/openurl?url%5Fver=Z39.88-2004&res%5Fdat=xri:pqdiss&rft%5Fval%5Ffmt=info:ofi/fmt:kev:mtx:dissertation&rft%5Fdat=xri:pqdiss:MQ99354
TICHÝ, Vlastimil. "Charakterizace signální dráhy adenosinu v buňkách imaginálních terčků \kur{Drosophila melanogaster}." Master's thesis, 2007. http://www.nusl.cz/ntk/nusl-46707.
Повний текст джерелаYang, Yi Chen, and 楊伊琛. "The Relationship between the Catalytic Cycle and Structure of Dopamine N-Acetyltransferase from Drosophila melanogaster." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/06424457062033037646.
Повний текст джерела國立清華大學
生物資訊與結構生物研究所
104
Dopamine N-acetyltransferase (Dat) found in Drosophila melanogaster belongs to arylalkylamine N-acetyltransferase (AANAT, EC 2.3.1.87) family, which is a member of GCN5-related N-acetyltransferase (GNAT) superfamily. Dat catalyzes arylalkylamine N-acetylation which transfers acetyl group of acetyl-CoA (Ac-CoA) to arylalkylamine to generate N-acetyl-arylalkylamine and CoA. AANAT had been reported the ordered bi bi sequential mechanism by enzyme inhibition analysis as well. In our previous study, we had also determined Dat is ordered bi bi sequential mechanism using isothermal titration calorimetry (ITC) and enzyme inhibition kinetics. Dat has to bind cofactor (Ac-CoA) first and then followed by substrate (Arylalkylamine). Nevertheless, the underlying structural mechanism still remains ambiguous. Furthermore, we had found the electron density map of products, N-acetyl-arylalkylamine and CoA, on substrate and cofactor binding site in ternary structure by soaking. It seemed products cannot auto-release. Thus, we interested in how Dat conducts enzymatic recycling and the catalytic process. In this study, we solved 1.20 Å resolution ternary structure (Dat/N-acetyl-arylalkylamine/CoA) of Dat by co-crystallization. Comparing apo form, binary form (tDat/Ac-CoA complex) and ternary form (tDat/CoA/Ac-PEA complex) of tDat, we found conformation of apo form Dat was different from binary and ternary form among them; structures of binary and ternary form were similar with each other. Then, we found the conformational change after Ac-CoA binding with tDat. The conformational change of substrate binding site may decide whether tDat can binding substrate or not. Thus, we elucidated ordered bi bi sequential mechanism of Dat by x-ray structural analysis. Additionally, the overall ternary structure of co-crystallization was similar with soaking except for an additional Ac-PEA outside the protein surface. The phenomenon implied may exist some factors to facilitate product release and enzyme recycle of Dat. Using isothermal titration calorimetry (ITC) and x-ray co-crystallization to carry out the competitive experiments, we found Ac-CoA showed dominantly competitive relation with respect to CoA. The dominantly competitive relation between Ac-CoA and CoA may resulted in enzymatic recycle of Dat. Finally, we based on catalytic triad to generate three variants, E47D, E47Q and E47N, for approaching transition state. All of them lost their substrate binding affinity and led to dramatic catalytic activity decrease. According to our results, we suggested Ac-CoA priorly binds to Dat providing a conformation change which facilitates substrate binding and forms ternary form. Ac-CoA may serve as driving force in catalyzation process of Dat. Our results implied Ac-CoA drives CoA and N-acetyl-arylalkylamine in Dat (ternary form) away, and occupies the cofactor binding site of Dat. Then Dat returns to state of binary form and accomplishes enzymatic recycling.
Coban-Poppinga, Büsra. "Plasticity of Dopamine-Releasing Central Brain Neurons Underlying Adaptational Feeding-Related Behavior in Drosophila Melanogaster." Doctoral thesis, 2020. http://hdl.handle.net/21.11130/00-1735-0000-0005-138E-6.
Повний текст джерелаSantoso, Clarissa Stephanie. "Analysis of the ABC transporter CG31731 in engulfment during programmed cell death in the Drosophila melanogaster ovary." Thesis, 2017. https://hdl.handle.net/2144/31673.
Повний текст джерелаDing, Wei Cheng, and 丁韋丞. "Study of the ordered sequential mechanism and critical residues in binding site of Drosophila melanogaster dopamine N-acetyltransferase." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/74011879411272799834.
Повний текст джерела國立清華大學
生物資訊與結構生物研究所
104
Dopamine is a neurotransmitter and associated with many physiological mechanisms, for example, cognition, circadian rhythm, aging, memory and learning. Dopamine N-acetyltransferase (Dat), an arylalkylamine N-acetyltransferase (AANAT), is identified in Drosophila melanogaster involves in the catabolism of monoamines and sclerotization. Dat transfers acetyl group from acetyl coenzyme A (AcCoA) to arylalkylamine and produces N-acetylarylalkylamine. Previous study in our laboratory revealed that Dat obeyed an ordered sequential mechanism: AcCoA binding first, substrates binding afterward, and then acetyl group transferring. In order to understand the detail of this catalysis process, isothermal titration calorimetry (ITC) was used to study the thermodynamics changes and binding affinities in reactions. To confirm the ordered sequential mechanism, sequential addition of cofactors and substrates into 15N labeled Dat protein was performed and monitored by nuclear magnetic resonance (NMR), and then the chemical shift changes of 15N heteronuclear single quantum coherence (HSQC) spectra were analyzed. ITC results were consistent with NMR results and proved the existence of ordered sequential mechanism. Recently, ternary structure which was co-crystalized by Dat, AcCoA, and substrate was resolved by our laboratory. However, this ternary structure was consisted of Dat, CoA, and acetyl-substrate. This indicated that final products did not release from Dat after catalysis. To further investigate how the products leave Dat and let the next reaction occur. Titration of AcCoA into Dat containing products was monitored by ITC, and the formation of new products was confirmed by DTNB assay. The results showed that AcCoA replaced CoA in Dat, and further let acetyl-substrate leave. Then a new Dat-AcCoA complex was ready to bind a new substrate and transfer acetyl group to it. To explore residues affecting cofactor or substrate binding, Ligplot+ was used to analyze the binary form of Dat structure (PDB code: 3TE4) and ternary form (unpublished). In Ligplot+ analysis report, R153 and K192 are related with AcCoA/CoA binding, while M121 may participate in substrate binding. In CAVER software analysis, M121 and D142 located in the narrowest of the substrate binding tunnel of Dat. Four residues were replaced to alanine, and ITC and enzyme activity assay was used to check their roles. Unexpectedly, M121A totally lost the AcCoA binding ability in ITC test, and remained only 13% of activity in functional assay. Considering the notable changes in secondary structure of M121A and far distance between M121 and cofactor, the loss of cofactor binding ability of M121A should be caused from the change in structure rather than the interaction with cofactor. R153 was important residue for AcCoA binding because no cofactor binding was detected in ITC test of R153A. K192A showed less binding affinity to AcCoA/CoA in ITC test, but remained its catalytic ability. D142A showed no significant effects on binding with cofactor or substrate, and on catalysis. In conclusion, Dat obeyed an ordered sequential mechanism: AcCoA binding first, substrates binding afterward, and then acetyl group transferring. To start next reaction, AcCoA replaced CoA in Dat, and further let acetyl-substrate leave. Then a new Dat-AcCoA complex was ready to bind a new substrate and transfer acetyl group to it. Besides, M121 affected on protein structure and R153 was important residue participating in cofactors binding to Dat.
Marcos, Joana Teixeira de Melo Palma. "Regulação do mate-choice copying por neurónios apetitivos ou neurónios aversivos de dopamina em Drosophila melanogaster." Master's thesis, 2019. http://hdl.handle.net/10400.12/7494.
Повний текст джерелаO presente estudo descreve o papel de neurónios específicos de dopamina na regulação da aprendizagem social, em contexto sexual, em Drosophila melanogaster. A dopamina (DA) é um neuro-modulador que controla a formação de memórias associativas, com valência positiva (memórias apetitivas) ou negativa (memórias aversivas) durante o processo de aprendizagem associal, e a nossa pergunta é se o mesmo mecanismo também ocorre na aprendizagem social. Como paradigma de aprendizagem social, utilizámos o mate-choice copying (MCC), que é uma forma de aprendizagem social por observação das interações sexuais entre conspecíficos, e que se sabe ser regulado pela dopamina. Como controlo, utilizámos o teste de independent mate choice (IMC), que avalia a preferência inata das fêmeas por machos com fenótipos distintos, neste caso fenótipos artificiais de cor azul e rosa. Replicámos estes dois paradigmas comportamentais num grupo controlo (uma linha heterozigótica, a Canton-Special, ou CS), e em duas linhas transgénicas com neurónios dopaminérgicos desativados: a linha PAM para os neurónios apetitivos, e a linha PPL1 para os neurónios aversivos. Os resultados mostram que a linha CS faz MCC, a linha PAM não faz e a PPL1 parece fazer, mas este último resultado não é conclusivo, provavelmente porque o tamanho da amostra é pequeno. Sabendo que as memórias apetitivas são importantes nos processos motivacionais, incluindo a motivação para o acasalamento, a formação de memórias apetitivas durante o MCC é certamente um mecanismo adaptativo. Por essa razão, poderá haver um menor contributo das memórias aversivas, contudo, o nosso estudo deixa essa hipótese ainda em aberto.
The present study describes the role of dopamine-specific neurons in regulating social learning in a sexual context, in Drosophila melanogaster. Dopamine (AD) is a neuro-modulator that controls the formation associative memories, with positive (appetitive) or negative (aversive) valence during the asocial learning process, and our question is whether the same mechanism also occurs in social learning. As paradigm of social learning, we used mate-choice copying (MCC), which is a form of social learning by observing conspecific sexual interactions, and that is known to be regulated by dopamine. As a control, we used the independent mate choice (IMC) test, which assesses female innate preference for males with distinct phenotypes, in this case the artificial blue and pink phenotypes. We replicated these two behavioural paradigms in a control group (a heterozygous line, Canton-Special, or CS), and in two transgenic lines with dopaminergic deactivated neurons: the PAM line for appetitive neurons and the PPL1 line for aversive neurons. The results show that the CS line does MCC, the PAM line does not, and the PPL1 line seems to do, but the latter result is not conclusive, probably because the sample size is small. Knowing that appetitive memories are important in motivational processes, including mating motivation, the formation of appetitive memories during MCC is certainly an adaptive mechanism. For this reason, there may be a minor contribut