Made available in DSpace on 2014-06-11T19:30:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-03-30Bitstream added on 2014-06-13T21:01:38Z : No. of bitstreams: 1 schneider_mc_dr_rcla.pdf: 2704348 bytes, checksum: d421ba97ff151da8a48de6dc098ff7cb (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo deste trabalho é caracterizar citogeneticamente 13 espécies da família Elateridae, pertencentes à subfamília Agrypninae, tribo Conoderini (Conoderus dimidiatus, Conoderus fuscofasciatus, Conoderus malleatus, Conoderus rufidens, Conoderus scalaris, Conoderus stigmosus, Conoderus ternarius e Conoderus sp.), subfamília Agrypninae, tribo Pyrophorini (Pyrearinus candelarius, Pyrophorus divergens e Pyrophorus puncatissimus), e subfamília Elaterinae, tribo Agriotini (Cardiorhinus rufilateris e Pomachilus sp.2), visando estabelecer as principais estratégias de diferenciação cromossômica que ocorreram nestas espécies. Através da análise de células testiculares meióticas foi possível notar que as 13 espécies de Elateridae apresentam cromossomos com comportamento regular durante a meiose e que o bivalente sexual neoXY de Conoderus stigmosus possui um quiasma terminal. As principais estratégias de diferenciação cromossômica detectadas nas espécies de Elateridae são apresentadas e discutidas neste trabalho.

La différenciation et le développement requièrent une régulation fine de l’expression desgènes, médiée en partie par les modifications épigénétiques. Parmi les modificationsd’histones, la chromatine bivalente, signature chromatinienne atypique associant lesmarques permissive H3K4me2/3 et répressive H3K27me3, est de par sa plasticité, pressentiepour jouer un rôle décisionnel dans l’acquisition d’une identité cellulaire. Pour étudier le rôlede la chromatine bivalente au cours du développement, nous avons choisi d’utiliserl’empreinte parentale. Ce cadre développemental bien caractérisé, conduit à l’expression decertains gènes à partir d’un seul des deux allèles selon son origine parentale. La méthylationdifférentielle de l’ADN d’une région clé, appelée ICR (Imprinting Control Region), bienqu’absolument requise pour l’expression mono-allélique de ces gènes, n’est pas suffisantepour rendre compte de la complexité du profil d’expression de ces gènes suggérantl’implication d’autres mécanismes. Sur 15 ICR méthylés sur l’allèle maternel, nous avonsprécisément mis en évidence que la chromatine bivalente est présente par défaut sur l’allèlenon-méthylé lorsque celui-ci est transcriptionnellement inactif, quel que soit le stadedéveloppemental ou le tissu étudié, participant ainsi à la régulation fine de l’expressiontissu-spécifique à partir de ces régions. Dans leur ensemble, nos données révèlent que lachromatine bivalente joue un rôle moins dynamique que pressentie. Ainsi, au niveau del’empreinte parentale, sa fonction principale serait de protéger l’allèle non-méthylé des ICRcontre l’acquisition de méthylation tout en aidant à le maintenir réprimé dans certainstissus. Nous proposons que la chromatine bivalente joue un rôle similaire sur l’ensemble desîlots CpG du génome, contribuant ainsi à la protection de l’identité cellulaire. Afin decompléter cette première étude, j’ai étudié la régulation de l’expression d’un candidat de larégulation de la dynamique de la chromatine bivalente, l’histone déméthylase pourH3K27me3, JMJD3. Les résultats obtenus suggèrent que l’induction d’expression observéeau cours de la différenciation neurale s’appuie sur une dynamique de la structuretridimensionnelle de la chromatine qui pourrait elle-même être régulée par la transcriptiond’un eARN (enhancer ARN) et l’hydroxyméthylation. Ce modèle souligne un mode derégulation complexe de ce nouvel acteur épigénétique, impliquant des régionsintragéniques, et pourrait notamment permettre de comprendre les mécanismes impliquésdans sa dérégulation dans les cancers. / Fine-tuned regulation of gene expression is required for cell fate determination anddevelopment. Epigenetics modifications are well documented to be instrumental in thisprocess. Among them, bivalent chromatin, an unusual chromatin signature, which associatesthe permissive mark H3K4me2/3 and the repressive mark H3K27me3, is believed to arbitrategene expression during cell commitment. To study its precise role in development, we haveundertaken to study bivalency in the context of genomic imprinting. This well-defineddevelopmental frame is a process restricting expression of some genes to one parental alleleonly. The constitutive differential DNA methylation at the key region called ICR (ImprintingControl Region), is absolutely required but not sufficient to explain the complexity of themono-allelic expression pattern of imprinted genes, indicating that other mechanisms couldbe involved. Specifically, on 15 maternally methylated ICR, we showed that bivalentchromatin is acquired by default on the unmethylated allele of ICR when it istranscriptionally inactive whatever the developmental stage or the tissue studied and thuscontribute to tissue-specific expression from these regions. Altogether, our results revealthat chromatin bivalency is much less dynamic than proposed. In the context of genomicimprinting, it seems to plays more a safeguard function at ICR by protecting theunmethylated allele against DNA methylation acquisition while keeping it silent in a subsetof tissues. To complete this study, I studied the regulation of JMJD3, a histone demethylasefor H3K27me3, candidate to regulate bivalency dynamic. Our results suggest that theinduction of Jmjd3 expression observed during neural differentiation rely on the dynamic ofthe tridimensional architecture at the locus which could be regulated by the transcription ofan eRNA (enhancer RNA) and by hydroxymethylation. This model highlight a complex way ofregulation for this new epigenetics actor, involving intragenic regions and could help tounderstand how Jmjd3 expression is deregulated in a pathological context such as in cancer.

As propriedades fotofísicas dos complexos morin-Mg2+ e morin-Ce3+, foram investigadas em solução metanólica, em filmes finos de polímeros e em nanozéolitas por técnicas de fluorescência. A associação do metal ao flavonoide ocorre no sítio da carbonila do anel pirona nas razões estequiométricas de 1:1 e 1:2 (magnésio: morin), e esta associação favorece no aumento considerável da intensidade de fluorescência devido à inibição da transferência intramolecular do próton no estado excitado. Os resultados revelaram que os espectros de emissão do complexo morin-Mg2+ sofreram um pequeno deslocamento hipsocrômico. Os decaimentos de fluorescência do sistema Morin-Mg2+ são triexponenciais para o complexos Morin-Mg2+ formados com tempo de vida máximo de 3 ns. O complexo morin-Ce3+ é pouco fluorescente e a complexação com o metal leva a supressão de fluorescência devido a sua natureza paramagnética. A estequiometria aparente dominante é de um 1:1 e o decaimento possui um componente de tempo de vida longo de 4-5 ns. Na comparação das propriedades de fluorescência do morin-Mg2+ com o indicador magnesium green verificamos que ambos são bons indicadores fluorescentes para a microscopia de fluorescência podendo esses complexos serem usados na detecção de Mg2+ em nanozeólita e outros sistemas / The photophysical properties of the molecular complexes of morin with Mg2+ and Ce3+ cations in methanol solution as well as in thin polymeric films and nanozeolytes have been investigated by using fluorescence spectroscopy and microscopy. The association of morin with Mg2+ occurs at the (4)-carbonyl group of the pyrone ring at 1:1 and 1:2 stoichiometric ratio. The complexes have shown an increase of the emission quantum yield by precluding the excited state deactivation though the intramolecular proton transfer process. The fluorescence decays of the complexes are non exponential due to the presence of more than one emission species and the average decay time found was about 3 ns. On the other hand, the addition of Ce3+ to morin solution resulted in fluorescence quenching ascribed to 1:1 non emissive complex due to the paramagnetic nature of the metal cation. The fluorescence properties of morin-Mg2+ complex have been compared with the standard fluorescence indicator magnesium green. The results pointed out that morin-Mg2+ may be used as an alternative low coast indicator of Mg2+ in homogenous as well as in microheterogeneous media.

As propriedades fotofísicas dos complexos morin-Mg2+ e morin-Ce3+, foram investigadas em solução metanólica, em filmes finos de polímeros e em nanozéolitas por técnicas de fluorescência. A associação do metal ao flavonoide ocorre no sítio da carbonila do anel pirona nas razões estequiométricas de 1:1 e 1:2 (magnésio: morin), e esta associação favorece no aumento considerável da intensidade de fluorescência devido à inibição da transferência intramolecular do próton no estado excitado. Os resultados revelaram que os espectros de emissão do complexo morin-Mg2+ sofreram um pequeno deslocamento hipsocrômico. Os decaimentos de fluorescência do sistema Morin-Mg2+ são triexponenciais para o complexos Morin-Mg2+ formados com tempo de vida máximo de 3 ns. O complexo morin-Ce3+ é pouco fluorescente e a complexação com o metal leva a supressão de fluorescência devido a sua natureza paramagnética. A estequiometria aparente dominante é de um 1:1 e o decaimento possui um componente de tempo de vida longo de 4-5 ns. Na comparação das propriedades de fluorescência do morin-Mg2+ com o indicador magnesium green verificamos que ambos são bons indicadores fluorescentes para a microscopia de fluorescência podendo esses complexos serem usados na detecção de Mg2+ em nanozeólita e outros sistemas / The photophysical properties of the molecular complexes of morin with Mg2+ and Ce3+ cations in methanol solution as well as in thin polymeric films and nanozeolytes have been investigated by using fluorescence spectroscopy and microscopy. The association of morin with Mg2+ occurs at the (4)-carbonyl group of the pyrone ring at 1:1 and 1:2 stoichiometric ratio. The complexes have shown an increase of the emission quantum yield by precluding the excited state deactivation though the intramolecular proton transfer process. The fluorescence decays of the complexes are non exponential due to the presence of more than one emission species and the average decay time found was about 3 ns. On the other hand, the addition of Ce3+ to morin solution resulted in fluorescence quenching ascribed to 1:1 non emissive complex due to the paramagnetic nature of the metal cation. The fluorescence properties of morin-Mg2+ complex have been compared with the standard fluorescence indicator magnesium green. The results pointed out that morin-Mg2+ may be used as an alternative low coast indicator of Mg2+ in homogenous as well as in microheterogeneous media.