O protozoário flagelado, Trichomonas vaginalis, causa a tricomonose, a doença sexualmente transmissível (DST) não viral mais comum no mundo. Metronidazol e tinidazol são os dois fármacos de escolha recomendados pelo Food and Drug Administration (FDA, USA) para o tratamento da tricomonose humana. Entretanto, isolados de T. vaginalis clínicos ou laboratoriais resistentes e reações adversas comuns têm sido reportados. Sabe-se que fungos produzem moléculas bioativas podendo ser uma fonte em potencial de novas moléculas antiparasitárias, sendo assim, a busca por compostos naturais bioativos em basidiomicetos pode ser uma abordagem interessante para descoberta de fármacos alternativos. O objetivo desse estudo foi investigar a presença, purificar e identificar substâncias com atividade anti-Trichomonas no meio de cultivo de micélios de basidiomicetos. Para a obtenção dos compostos bioativos, os basidiomicetos Amauroderma camerarium, família Ganodermataceae, e Gymnopilus pampeanus, família Cortinariceae, foram cultivados em meio caldo extrato de malte (CEM) com variação da fonte de nitrogênio e tempo de cultivo. O extrato bruto com maior atividade anti-T. vaginalis para A. camerarium (CEM + KNO3 28 dias de cultivo) foi escolhido para ensaios de purificação e caracterização, que indicaram que o composto bioativo é de natureza protéica. Uma proteína sem alta identidade com outra proteína foi purificada e foi denominada amaurocina. Amaurocina apresenta atividade contra isolados clínicos de T. vaginalis – TV LACH1 e TV LACM2 – (CIM= 62,4 μg/mL), sendo um isolado resistente ao metronidazol (TV LACM2), e contra o isolado ATCC 30236 (CIM= 31,2 μg/mL). A Amaurocina também apresenta atividade próinflamatória por induzir aumento da liberação de óxido nítrico de neutrófilos, o qual é um importamente mecanismo de defesa do hospedeiro durante a infecção parasitária. Para G. pampeanus, os extratos mais ativos foram produzidos em meio CEM + KNO3 por 28 dias de cultivo e CEM + KNO3 por 21 dias de cultivo, porém a purificação desses extratos é necessária. Esses resultados estão em concordância com o alto potencial de produção de biocompostos dos basidiomicetos, o qual tem sido reportado na literatura por décadas. / The flagellated protozoan, Trichomonas vaginalis, causes trichomonosis, the most common non-viral sexual transmitted disease (STD) in the world. Metronidazole and tinidazole are two drugs of choice recommended by Food and Drug Administration (FDA, USA) for treatment of human trichomonosis. However, clinical or laboratory-generated drug-resistant isolates of T. vaginalis and common adverse reactions have been reported. It is known that fungi produce bioactive molecules and they can be a potential source of new antiparasitic molecules. Thus, in order to improve the current chemotherapy of T. vaginalis infection, the search for natural bioactive compounds in basidiomycetes can be an interesting approach to discover alternative drugs. The aim of this study was to investigate the presence, to purify and to identify substances in cultivation of basidiomycetes’ mycelia, which have anti-Trichomonas activity. Mycelia of basidiomycete Amauroderma camerarium, family Ganodermataceae, and Gymnopilus pampeanus, family Cortinariceae, were transferred to flasks containing malt extract broth (MEB) medium with variation of the nitrogen source and time of growth. The preparation with higher anti-T. vaginalis activity (MEB + KNO3 28 days of growth) for A. camerarium basidiomycete was chosen for purification and characterization assays, that indicated that the bioactive compound is a protein-like molecule. A protein without high homology with any other protein was found and was named amaurocine. Amaurocine presents activity against clinical isolates of T. vaginalis – TV LACH1 and TV LACM2 – (MIC=62.4 μg/mL), one of these a metronidazole-resistant isolate, and it presents activity against the T. vaginalis ATCC 30236 isolate (MIC=31.2 μg/mL). Amaurocine also presents proinflamatory activity. Since it was able to enhance nitric oxide release from neutrophils, which is an important host defense mechanism during the parasitic infection. For G. pampeanus, the most active extracts were produced in MEB + KNO3 28 days of growth and MEB + KNO3 21 days of growth, but further steps of purification are necessary. These results are in agreement with the high potential of biocompounds production of basidiomycetes that has been reported in the literature for decades.

O protozoário flagelado, Trichomonas vaginalis, causa a tricomonose, a doença sexualmente transmissível (DST) não viral mais comum no mundo. Metronidazol e tinidazol são os dois fármacos de escolha recomendados pelo Food and Drug Administration (FDA, USA) para o tratamento da tricomonose humana. Entretanto, isolados de T. vaginalis clínicos ou laboratoriais resistentes e reações adversas comuns têm sido reportados. Sabe-se que fungos produzem moléculas bioativas podendo ser uma fonte em potencial de novas moléculas antiparasitárias, sendo assim, a busca por compostos naturais bioativos em basidiomicetos pode ser uma abordagem interessante para descoberta de fármacos alternativos. O objetivo desse estudo foi investigar a presença, purificar e identificar substâncias com atividade anti-Trichomonas no meio de cultivo de micélios de basidiomicetos. Para a obtenção dos compostos bioativos, os basidiomicetos Amauroderma camerarium, família Ganodermataceae, e Gymnopilus pampeanus, família Cortinariceae, foram cultivados em meio caldo extrato de malte (CEM) com variação da fonte de nitrogênio e tempo de cultivo. O extrato bruto com maior atividade anti-T. vaginalis para A. camerarium (CEM + KNO3 28 dias de cultivo) foi escolhido para ensaios de purificação e caracterização, que indicaram que o composto bioativo é de natureza protéica. Uma proteína sem alta identidade com outra proteína foi purificada e foi denominada amaurocina. Amaurocina apresenta atividade contra isolados clínicos de T. vaginalis – TV LACH1 e TV LACM2 – (CIM= 62,4 μg/mL), sendo um isolado resistente ao metronidazol (TV LACM2), e contra o isolado ATCC 30236 (CIM= 31,2 μg/mL). A Amaurocina também apresenta atividade próinflamatória por induzir aumento da liberação de óxido nítrico de neutrófilos, o qual é um importamente mecanismo de defesa do hospedeiro durante a infecção parasitária. Para G. pampeanus, os extratos mais ativos foram produzidos em meio CEM + KNO3 por 28 dias de cultivo e CEM + KNO3 por 21 dias de cultivo, porém a purificação desses extratos é necessária. Esses resultados estão em concordância com o alto potencial de produção de biocompostos dos basidiomicetos, o qual tem sido reportado na literatura por décadas. / The flagellated protozoan, Trichomonas vaginalis, causes trichomonosis, the most common non-viral sexual transmitted disease (STD) in the world. Metronidazole and tinidazole are two drugs of choice recommended by Food and Drug Administration (FDA, USA) for treatment of human trichomonosis. However, clinical or laboratory-generated drug-resistant isolates of T. vaginalis and common adverse reactions have been reported. It is known that fungi produce bioactive molecules and they can be a potential source of new antiparasitic molecules. Thus, in order to improve the current chemotherapy of T. vaginalis infection, the search for natural bioactive compounds in basidiomycetes can be an interesting approach to discover alternative drugs. The aim of this study was to investigate the presence, to purify and to identify substances in cultivation of basidiomycetes’ mycelia, which have anti-Trichomonas activity. Mycelia of basidiomycete Amauroderma camerarium, family Ganodermataceae, and Gymnopilus pampeanus, family Cortinariceae, were transferred to flasks containing malt extract broth (MEB) medium with variation of the nitrogen source and time of growth. The preparation with higher anti-T. vaginalis activity (MEB + KNO3 28 days of growth) for A. camerarium basidiomycete was chosen for purification and characterization assays, that indicated that the bioactive compound is a protein-like molecule. A protein without high homology with any other protein was found and was named amaurocine. Amaurocine presents activity against clinical isolates of T. vaginalis – TV LACH1 and TV LACM2 – (MIC=62.4 μg/mL), one of these a metronidazole-resistant isolate, and it presents activity against the T. vaginalis ATCC 30236 isolate (MIC=31.2 μg/mL). Amaurocine also presents proinflamatory activity. Since it was able to enhance nitric oxide release from neutrophils, which is an important host defense mechanism during the parasitic infection. For G. pampeanus, the most active extracts were produced in MEB + KNO3 28 days of growth and MEB + KNO3 21 days of growth, but further steps of purification are necessary. These results are in agreement with the high potential of biocompounds production of basidiomycetes that has been reported in the literature for decades.

Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-07-03T15:37:47Z No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-08T18:09:02Z (GMT) No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-08T18:13:13Z (GMT) No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5) / Made available in DSpace on 2015-07-08T18:28:04Z (GMT). No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5) Previous issue date: 2013-05-02 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / Mushrooms arouse interest for presenting high nutritional and medicinal value. The representatives of the genus Pleurotus are able to grow in different agricultural and agro-industrial waste, since they secrete numerous enzymes that hydrolyze the substrates prepared from waste providing nutrient The search for alternative substrates increased in recent years due to the availability of regional waste little or never used in the production of edible mushrooms. The use of waste in solid-state fermentation has emerged also in the production of enzymes. Proteases are a group of enzymes commercially exploited, however the use of Basidiomycetes as producers of proteases is still a relatively unexplored. The aim of this study was to evaluate the mycelial growth as well as to investigate the production and partially characterize extracellular proteases from Pleurotus ostreatoroseus DPUA 1720 in different agro-industrial waste, selecting a mixture for the production of the mushroom and check their microbiological quality and nutritional value. Cultures were prepared in potato dextrose added yeast extract 0.5% (w/v). The vertical mycelial growth and production of proteases from P. ostreatoroseus DPUA 1720 were performed in agro-industrial waste (acai seed, sawdust, cupuassu bark, rice bran, bark and pineapple crown) for 15 days under two growing conditions (presence and absence of light). The production of basidiomata was performed in the substrate which presented satisfactory mycelial growth and vigorous mycelium. From this bioprocess evaluated the formation and development of early basidiomata, total time of cultivation, biological efficiency, productivity, production rate, loss of organic matter, microbiological and nutritional value of the mushroom. P. ostreatoroseus DPUA 1720 grew in all industrial residues also tested serving as sources for the production of proteases. In bark cupuaçu added rice bran mycelium was strongly vigorous and the mycelial growth satisfactory, therefore selected for the production of P. ostreatoroseus DPUA 1720. Under the experimental conditions the proteolytic activity was determined in crude extracts from the substrates of vertical mycelial growth with maximum proteolytic activity (7.89 U/mL) in bark cupuassu added rice bran in the presence of light. This substrate was the most efficient for the production of proteases with activity atpH 6.0 and optimum temperature at 40 ˚C. The basidiomata showed no contaminants, low in fat, high in fiber and protein, macro and micro minerals, essential and non-essential amino acids, may therefore be inserted into the feed as a safe and nutritious food. / Os cogumelos despertam interesse por apresentar alto valor nutricional e medicinal. Os representantes do gênero Pleurotus são capazes de crescer em diferentes resíduos agrícolas e agroindustriais, pois secretam inúmeras enzimas que hidrolisam os substratos elaborados a partir dos resíduos disponibilizando nutrientes. A busca por substratos alternativos cresceu nos últimos anos em virtude da disponibilidade de resíduos regionais pouco ou nunca utilizados na produção de cogumelos comestíveis. O uso de resíduos na fermentação semi-sólida vem despontando também na produção de enzimas. As proteases são o grupo de enzimas mais exploradas comercialmente, no entanto o uso de Basidiomicetos como produtores de proteases ainda é um tema pouco explorado. O objetivo deste trabalho foi avaliar o crescimento micelial, bem como investigar a produção e caracterizar parcialmente proteases extracelulares de Pleurotus ostreatoroseus DPUA 1720 em diferentes resíduos agroindustriais, selecionar uma mistura para produção dos basidiomas e verificar sua qualidade microbiológica e valor nutricional. As culturas foram preparadas em ágar batata dextrose acrescido de extrato de levedura 0,5% (p/v). O crescimento micelial vertical e a produção de proteases de P. ostreatoroseus foram realizados em resíduos agroindustriais (semente de açaí, serragem, casca de cupuaçu, farelo de arroz, casca e coroa de abacaxi) durante 15 dias, sob duas condições de cultivo (presença e ausência de luz). A produção do basidioma foi realizada no substrato que apresentou crescimento micelial satisfatório e micélio vigoroso. A partir desse bioprocesso foi avaliada a formação e desenvolvimento dos primórdios, tempo total de cultivo, eficiência biológica, produtividade, taxa de produção, perda de matéria orgânica, qualidade microbiológica e valor nutricional dos basidiomas. P. ostreatoroseus DPUA 1720 cresceu em todos os resíduos agroindustriais testados, também servindo como fontes para a produção de proteases. Em casca de cupuaçu adicionado de farelo de arroz o micélio foi fortemente vigoroso e o crescimento micelial satisfatório, por isso selecionado para produção de P. ostreatoroseus DPUA 1720. Nas condições experimentais a atividade proteolítica foi determinada nos extratos brutos provenientes dos substratos miceliados do crescimento micelial vertical com máxima atividade proteolítica (7,89 U/mL) em casca de cupuaçu adicionado de farelo de arroz na presença de luz. Este substrato foi o mais eficiente para a produção das proteases com atividade em pH 6,0 e temperatura ótima a 40˚C. Os basidiomas apresentaram ausência de contaminantes, baixo teor de lipídios, alto teor de fibras e proteínas, macro e microminerais, aminoácidos essenciais e não essenciais, podendo assim ser inserido na alimentação como um alimento saudável e nutritivo.

Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-10-23T19:44:00Z No. of bitstreams: 2 Dissertação - Evandro Leite Bitencourt - 2015.pdf: 4223075 bytes, checksum: fe60d827b473e79c5c9c8ab601f67a69 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-10-23T19:46:10Z (GMT) No. of bitstreams: 2 Dissertação - Evandro Leite Bitencourt - 2015.pdf: 4223075 bytes, checksum: fe60d827b473e79c5c9c8ab601f67a69 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-10-23T19:46:10Z (GMT). No. of bitstreams: 2 Dissertação - Evandro Leite Bitencourt - 2015.pdf: 4223075 bytes, checksum: fe60d827b473e79c5c9c8ab601f67a69 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-03-31 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Mushrooms have been valued as a functional food and dietary supplement for humanity, due to the presence of substances with high nutritional value and/or therapeutic. Among the mushrooms recognized for its therapeutic properties we have is the leafy Grifola, popularly known as “Maitake”, which has important pharmacological actions such as anti-tumor, immunomodulatory, antiinflammatory, antioxidant, and others, which have been mainly related to its carbohydrates. Nevertheless, this study aimed to review the chemistry of the polysaccharides from mushroom of the two lines ("white" and "gray", represented as Gfb and Gfc, respectively) of G. frondosa leafy that have been cultivated in Brazil. From these type it was obtained branched -glucans containing links of type (13) (16) and heteropolysaccharides formed mainly by galactose (heterogalactan) or mannose (heteromannan). The heterogalactan composed mainly of fucose (15.7%), mannose (20.5%) and galactose (63.8%) was denominated fucomannogalactan (FMG-Gfb). It contains a main chain consisting of units of α-D-Galp and 3-Me-O-α-D-Galp connected (16), which are present partially substituted by O-2 disaccharide 3-O- α-D-Manp-α-L-Fucp to a lesser extent with terminal non-reducing α-L-Fucp and α-D-Manp. The heteromannan composed of fucose (6.9%), xylose (33.1%) and mannose (60.0%), a fucoxylomannan (FXM-Gfc) was found to be composed of a backbone of units α-D-Manp (13) connected, and a part of them substituted by O-3 side chains made of fucose and xylose. The results of this study suggest that there are no significant differences between the carbohydrate of both strains, showing that white strain it provide the therapeutic effects attributed to carbohydrates of another strain. / Os cogumelos têm sido valorizados como alimento funcional e suplemento alimentar para a humanidade, devido à presença de substâncias com alto valor nutricional e/ou medicinal. Dentre os cogumelos reconhecidos pelas suas propriedades terapêuticas encontra-se o Grifola frondosa, popularmente conhecido como Maitake, o qual possui importantes ações farmacológicas como antitumoral, imunomoduladora, anti-inflamatória, antioxidante, entre outras, as quais têm sido relacionadas, principalmente, aos seus carboidratos. Diante do exposto, o presente trabalho teve como objetivo a avaliação química dos polissacarídeos dos basidiomas das duas linhagens (branca e cinza, representadas como Gfb e Gfc, respectivamente) de G. frondosa que vem sendo cultivadas no Brasil. A partir destes foram obtidas as -glucanas ramificadas contendo ligações do tipo (13) (16) e os heteropolissacarídeos formados principalmente por galactose (heterogalactana) ou manose (heteromanana). A heterogalactana constituída, principalmente, por fucose (15,7%), manose (20,5%) e galactose (63,8%), foi denominada de fucomanogalactana (FMG-Gfb). Esta contém uma cadeia principal formada por unidades de α-D-Galp e de 3-O-Me-α-D-Galp ligadas (16), as quais se apresentam parcialmente substituídas em O-2 pelo dissacarídeo 3-O-α-DManp- α-L-Fucp e em menor proporção com terminais não redutores de α-LFucp e α-D-Manp. A heteromanana, composta por fucose (6,9%), xilose (33,1%) e manose (60,0%), ou seja, uma fucoxilomanana (FXM-Gfc) mostrou ser constituída por uma cadeia principal composta por unidades de α-D-Manp (13) ligadas, sendo uma parte destas substituídas em O-3 por cadeias laterais formadas por fucose e xilose. Os resultados obtidos neste estudo sugerem que não há diferenças significativas entre os carboidratos de ambas as linhagens, evidenciando que a linhagem branca deve apresentar os efeitos terapêuticos atribuídos aos carboidratos da outra linhagem.

Saprophytic fungi can be paired with companion crops in interplant systems to increase production efficiency. However, fungal species/strain, substrate, and inoculation rate can affect the growth of companion crops. This project investigated the viability of open-field mushroom production by interplanting three strains of Pleurotus ostreatus (Elm A, Elm B, and 8801) with kale (B. oleracea var. acephala) and forage radish (Raphanus raphanistrub sub. sativus), and measured the effect of interplanting on plant yield over two field seasons. In the field, Elm A showed an increase in plant yield at a low inoculation rate and decrease in plant yield at a high inoculation rate, compared to the untreated. Conversely, 8801 showed a reduction in plant yield at high and low inoculation rates in the field. Elm B at a high rate showed a reduction in plant yield both in the field and greenhouse. Kale was grown in hydroponics with fungal secretions added at a range of concentrations (10, 100, 1,000 and 10,000 ppm). Elm A showed an overall increase in plant yield in hydroponics, and Elm B showed an overall decrease in plant yield, compared to the untreated. Mushroom production was low in field plots and was not a commercially viable option. Pleurotus ostreatus interplanting methods with companion crops need improvement to make this a commercially viable practice.

This thesis deals with microstrip antennas covered by the metamaterials. First, are described planar antennas, their problems and the emergence of surface waves. Surface waves can cause unwanted coupling among particular parts of the structure and can degrade its parameters. The problem can be solved using an electromagnetic band gap structure (EBG). These periodic structures are able to suppress surface waves in different frequency bands. It is shown how the EBG structure in the function superstate improve directivity and antenna gain. Radiation conventional microstrip antenna with metallo-dielectric EBG superstrate and with the purely dielectric double-layer superstrate is described. The both structures are designed and simulated in CST Microwave Studio program. Further is described the antenna radiation with so-called mushroom structure and metallo-dielectric EBG superstate. The structure is again designed and simulated in CST MWS program. Finally, there are two structures with metallo-dielectric superstate implemented and measured.

Les lectines sont des protéines ubiquitaires qui se lient spécifiquement et de manière réversible aux sucres sans les modifier. Elles peuvent déchiffrer le glycocode : les informations stockées dans les sucres à la surface d'une cellule. Les lectines sont impliquées dans de nombreux processus biologiques tels que la cohésion cellulaire, le développement, la signalisation cellulaire, la défense ou les infections microbiennes. Elles peuvent présenter des propriétés exploitables, par exemple antiprolifératives, antivirales ou antitumorales et sont des outils recherchés en glycobiologie. Le règne fongique a suscité un large intérêt ces dernières années, car il est une source prometteuse et largement inexplorée pour les lectines ayant de nouvelles spécificités et/ou activités. Les données structurales sont encore limitées pour les lectines fongiques et ce projet vise notamment à combler cette lacune en étudiant deux familles de lectines reconnaissant des marqueurs glycosidiques associés aux cancers. Via une approche multidisciplinaire, l'interaction de ces lectines avec leur ligand au niveau atomique a été disséquée et des modifications ont été réalisées pour les rendre plus spécifiques pour un motif particulier. Les lectines recombinantes sauvages ou mutées ont été exprimées dans Escherichia coli puis purifiées par des méthodes chromatographiques classiques. La spécificité et l'affinité ont été déterminées par différentes techniques telles que l'hémagglutination, les puces à sucre et la microcalorimétrie de titration. La caractérisation structurale a été effectuée par cristallographie aux rayons X et a permis de définir les déterminants de la protéine et du ligand indispensables à l'interaction et d'aider à la conception et à l'évaluation structurale de mutants. Enfin, des analyses sur cellules et tissus cancéreux ont confirmé leur potentiel en tant qu’outils de marquage, ce qui permet d’envisager leur utilisation pour des applications biomédicales ou biotechnologiques. / Lectins are ubiquitous proteins which bind specifically and reversibly carbohydrates without modifying them. They can decipher the glycocode: information stored in carbohydrates found at the surface of any cell. Lectins are involved in many biological processes such as cell cohesion, development, cell signaling, defense or microbial infections. They can present exploitable properties such as antiproliferative, antiviral or antitumor and are interesting tools in glycobiology. The fungal kingdom has attracted wide interest in the recent years since it is a promising and largely unexplored source for lectins for novel specificity and activities. Structural data are still limited for fungal lectins and this project aims to fill this gap of knowledge whilst studying new lectins specific for two glycosidic cancer markers. Via a multidisciplinary approach, we dissected the interaction of those lectins with their ligand at the atomic level or modified them to optimized their specificity or affinity. Recombinant wild-type and mutated lectins were expressed in Escherichia coli and then purified by classical chromatography methods. The specificity and affinity were determined by different techniques such as hemagglutination, glycan arrays, and microcalorimetry. Finally, structural characterization was performed by X‐ray crystallography in order to localize the sugar binding site, to obtain the specific protein and ligand determinants necessary for the interaction and to help for the design and structural evaluation of mutants. Finally, analysis on cancer cells and tissues confirmed their potential as labelling tools thereafter for biomedical or biotechnological applications.

No description available.

A functional central nervous system (CNS) is composed of numerous types of neurons. Neurons are derived from a limited number of multipotent neural stem cells. Previous studies have suggested three major strategies nature uses to diversify neurons: lineage identity specification that gives an individual neural stem cell distinct identity based on its position in the developing CNS; temporal identity specification that gives neurons derived from a neural stem cell distinct identities based on their birth-order within the lineage; and binary cell fate specification that gives different identities to the two sister postmitotic neurons derived from the terminal division of a common precursor. Through the combination of the three strategies, almost unlimited neuron types can be generated. To understand neuronal diversification, we have to understand the underlying molecular mechanisms of each of the three strategies. The fruit fly Drosophila melanogaster, has been an excellent model for studying neuronal diversity, mainly due to its easily traceable nervous system and an impressive collection of genetic tools. Studies in fly have provided us fundamental insights into lineage identity, temporal identity, and binary cell fate specifications. Nevertheless, previous studies mostly centered on the embryonic ventral nerve cord (VNC) because of its simpler organization. Our understanding of the generation of neuronal diversity in the fly brain is still rudimentary. In this thesis work, I focused on the mushroom body (MB) and three antennal lobe neuronal lineages, studying their neuronal diversification during postembryonic brain development. In Chapter I, I reviewed the previous studies that have built our current understanding of the neuronal diversification. In Chapter II, I showed that MB temporal identity changes are instructed by environmental cues. In Chapter III, to search for the potential factors that mediate the environmental control of the MB temporal identity changes, I silenced each of the 18 nuclear receptors (NRs) in the fly genome using RNA interference. Although I did not identify any NR important for the regulation of MB temporal identities, I found that unfulfilled is required for regulating axon guidance and for the MB neurons to acquire all major subtype-specific identities. In Chapter IV, I demonstrated that the Notch pathway and its antagonist Numb mediate binary cell fate determination in the three classical antennal lobe neuronal lineages— anterodorsal projection neuron (adPN), lateral antennal lobe (lAL), and ventral projection neuron (vPN)—in a context-dependent manner. Finally, in Chapter V, I did detailed lineage analysis for the lAL lineage, and identified four classes of local interneurons (LNs) with multiple subtypes innervating only the AL, and 44 types projection neurons (PNs) contributing to olfactory, gustatory, and auditory neural circuits. The PNs and LNs were generated simultaneously but with different tempos of temporal identity specification. I also showed that in the lAL lineage the Notch pathway not only specifies binary cell fates, but is also involved in the temporal identity specification.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第20421号 / 農博第2206号 / 新制||農||1047(附属図書館) / 学位論文||H29||N5042(農学部図書室) / 京都大学大学院農学研究科森林科学専攻 / (主査)教授 吉村 剛, 教授 梅澤 俊明, 教授 本田 与一 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM