Inbreeding can have profound negative effects on individuals by reducing fertility and viability. In populations, inbreeding depression can reduce growth rates and increases extinction risk. The aims of this thesis are to investigate inbreeding depression in male guppies (Poecilia reticulata) and to study the evolution of mechanisms for inbreeding avoidance in females, using guppies from a feral population in Queensland, Australia. Male guppies are highly polymorphic in their sexual ornamentation, indeed they show one of the most extreme polymorphisms observed in nature. Female guppies exhibit complex mate choice based on preferences for ornamentation, as well as social context. I aim is to examine how these factors of inbreeding avoidance alter sexual selection. In male guppies I found strong inbreeding depression in male sperm numbers, which is amplified under semi-natural compared to laboratory conditions (Chapter 2). Moreover, inbreeding depression results in low fertility under sperm competition: an experiment using artificial insemination techniques reveals that highly inbred males are heavily disadvantaged in gaining paternity (Chapter 3). On population level, inbreeding depression is manifest in reduced growth rates, predominantly in the early stages of inbreeding (Chapter 4). Population growth at inbreeding coefficients f=0.375-0.59 did not seem to lead to inbreeding depression, whereas lower levels of inbreeding reduced population growth. Although the growth rates in inbred populations appear normal, severe inbreeding depression is uncovered after outbred immigrants are added. Specifically, male immigrants are most efficient in short-term genetic rescue, probably due to insemination of large numbers of females whereas females are limited in the number of eggs they can produce (Chapter 4). Male ornamental traits show significant inbreeding depression in semi-natural conditions only (Chapters 2 & 3). Inbreeding avoidance mechanisms seem to have evolved in females: they prefer courtship displays of non-inbred males (Chapter 2), unfamiliar males (Chapter 5) and males with rare patterns (Chapter 6). This preference might increase the mating success of immigrants, and may have evolved to facilitate the avoidance of inbreeding. Together with context-independent preferences for ornament combinations (Chapter 6), it also offers an explanation for the maintenance of polymorphism in this species.

I investigated the effect of environmental factors (temperature and food availability) on the development and growth of semiaquatic bug Microvelia reticulata Burmeister, 1835. I focused on the developmental rate isomorphy. In a cross-designed experiment, M. reticulata was individually kept over whole juvenile period (eggs to reaching maturity) and subjected to treatments of three temperature (17, 21, 25 °C) and two food availability (full - daily feeding and reduced - 2 days interval) and their interaction. Development and growth was strongly influenced by temperature. Longest postembryonic development measured in individuals kept in the lowest temperature (54 days) contrasted to those kept in the highest temperature (16 days). Mortality was also significantly influenced by temperature; highest mortality rate was found in the 17 °C treatment. Effect of food availability was significant only in low temperature, resulting in the better survival of individuals on the high food level. In other temperature treatments, I found no effect of food on survival rate of M. reticulata. However, the food level influenced development significantly. In the 21 °C and 25 °C treatment, individuals on a full food developed faster than individuals on a reduced food. In 17 °C the food availability had no effect on time required to reach maturity. The best fitness (expressed as combination of low mortality rate and short development) was observed in 21 °C, both other temperature treatment were found stressing. Food availability had stronger effect on female individuals compared to males. In conclusion, I found no evidence of developmental rate isomorphy in M. reticulata. The research was supported by project of the Grant Agency of the Czech Republic GAČR P505/10/0096.

Wing polymorphism of semiaquatic bugs is affected by environmental factors as temperature, photoperiod, population density or permanency of habitat. The heritability has its own specific part, too. A lot of field studies suggest that crucial effect on wing polymorphism has temperature, but laboratory experiments often do not confirm this hypothesis. The main aim of this thesis was to describe the effect of environmental factors (photoperiod, temperature and permanency of habitat) on wing polymorphism of Microvelia reticulata, additionally of Gerris lacustris. The partial tasks included description of effect of mentioned abiotic factors on mortality and rate of development. The individuals of both species were kept at combinations of two levels of temperature, photoperiod and permanency of habitat (overall at eight treatments) from first larval instar to adult. All adults of Microvelia reticulata emerged as apterous, so there was no response of wing polymorphism to tested environmental factors. The main influence of heritability is presumable. Only ten individuals of G. lacustris developed into the adult stage, so it was impossible to describe the influence of abiotic factors on wing polymorphism, mortality and lenght of development in this species. Mortality of individuals of M. reticulata was significantly influenced by the type of substrate. Only 3.5% of individuals survived when reared on wet filter paper, while 80.5% on the water surface. Both other factors (temperature, photoperiod) had no effect on total mortality. There was no difference between mortality of males and females, so it is impossible to note sexual dependent mortality. The lenght of development did not differ between sexes either. Lenght of development was significantly affected by temperature, as low temperature reduces development of M. reticulata. The fastest development showed nymphs of second instar and slowest development nymphs of fifth instar at all treatments. Since the effect of abiotic factors on wing polymorphism was not found in M. reticulata, it would be sufficient to study the effect of heritability on development of wings in this species in the future.

The invasion of individuals into new habitats can pose a major threat to native species and to biodiversity itself. However, the consequences of invasions for native populations that are not fully reproductively isolated from their invaders are not yet well explored. Here I chose the Trinidadian guppy, Poecilia reticulata, to investigate how different population traits shaped the outcome of Haskins's introduction, a well-documented invasion of Guanapo river guppies into the Turure river. I especially concentrated on the importance of behaviour for invasive success. I investigated if the spread of Guanapo guppies is due to superiority in behaviour, life-history and/or genetics, or if the outcome of this translocation is due to chance. Despite the fact that by today the invasive front has passed the Turure's confluence with the River Quare many kilometres downstream of the introduction site, and the original genotype only survives in small percentages, as was revealed by genetic analysis in this and other studies, no obvious differences between invasive and native populations could be detected in any of the tested behavioural, life-history and genetic traits. When tested for mate choice, neither Guanapo nor Oropuche (Turure) males seemed to be able to distinguish between the population origin of females, but courted and mated at random. At the same time, females did not prefer to school with individuals of the same population over schooling with more distantly related females. The formation of mixed schools after an invasive event is therefore likely. Because female guppies showed a very low willingness to mate, even after having been separated from males for up to six months, sperm transfer through forced copulations will become more important. Taken together, these behaviours could increase the speed of population mixing after an invasion without the need for behavioural superiority of the invasive population. When tested for their schooling abilities, offspring of mixed parentage, in contrast to pure breds, displayed a large amount of variety in the time they spent schooling, a circumstance that can potentially influence survival rates and therefore the direction of gene pool mixing. Guanapo fish did not show reproductive superiority in a mesocosm experiment, where both populations were mixed in different proportions. On the contrary, in two out of three mixed treatments, the amount of Oropuche (Turure) alleles was significantly higher than expected from the proportion of initially stocked fish. The almost complete absence of distinguishable traits other than genetic variation between the examined populations that belong to different drainage systems, opposes the recent split of the guppy into two different species following drainage system borders, as is argued in this thesis. However, the successful invasion of the Turure by Guanapo guppies and the nearly entire disappearance of the original population can be explained in absence of differing population traits. Here I demonstrate how behavioural and genetic interactions between subspecies influence the outcome of biological invasions and second, how factors other than population traits, such as the geographic situation, can produce an advantageous situation for the invader even in the absence of population differences.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-08-25T14:53:55Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DesempenhoGuppyPoecilia.pdf: 4012755 bytes, checksum: e38444c9cd0f96e294f8f50246678089 (MD5) / Approved for entry into archive by Irvana Coutinho (irvana@ufpa.br) on 2017-08-28T17:33:16Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DesempenhoGuppyPoecilia.pdf: 4012755 bytes, checksum: e38444c9cd0f96e294f8f50246678089 (MD5) / Made available in DSpace on 2017-08-28T17:33:16Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DesempenhoGuppyPoecilia.pdf: 4012755 bytes, checksum: e38444c9cd0f96e294f8f50246678089 (MD5) Previous issue date: 2016-09-23 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O uso de modelos animais em pesquisa experimental nas últimas décadas tem se mostrado mais diversificado do que o modelo clássico através do uso de roedores ou primatas. Isso se dá principalmente pelos avanços nos estudos moleculares, morfológicos e funcionais que revelaram uma grande homologia entre os vertebrados. Nesta perspectiva, o peixe Zebrafish (Danio rerio) tem se mostrado como o animal não-mamífero com maior ascensão como animal de estudo em ciências biológicas nas últimas décadas. Entretanto, outras espécies de peixes também se mostram promissoras como alternativas de uso como modelo animal. Este estudo utilizou o Guppy (Poecilia reticulata) como modelo para pesquisa em comportamento através de diferentes abordagens experimentais. No estudo I, guppies foram expostos e reexpostos em diferentes turnos (manhã, tarde, noite e madrugada) nos testes de campo aberto e preferência claro-escuro. Os resultados encontrados mostram que em ambos os testes, machos e fêmeas apresentam diferenças comportamentais, sendo sensíveis a reexposição, com capacidade de aprendizagem e controle do ciclo circadiano. No estudo II, foi desenvolvido um labirinto em cruz com rampa no qual se verificou a sensibilidade da espécie ao aparato, o perfil de resposta mediante reexposição e o efeito de drogas. Os resultados revelaram sensibilidade ao aparato para uma altura de coluna d’água de 8cm e 5 minutos de sessão e diferenças entre os sexos e aprendizagem por habituação ao longo das reexposições. O estudo farmacológico indica que neste aparato a espécie é sensível a drogas ansiolíticas e ansiogênicas. Ao final, pode-se concluir que o Guppy apresenta comportamento similar e respostas as drogas compatível com os dados descritos para Zebrafish. Tais similaridades reforçam o uso de peixes como uma alternativa ao uso de mamíferos na experimentação animal. / The use of animal models in experimental research in the last decades has shown to be more diversified than the classical model through the use of rodent or primate. This is mainly by advances in molecular, morphological and functional studies that revealed a big homology between the vertebrates. On this perspective, the Zebrafish (Danio rerio) has shown to be the non-mammal animal with biggest ascension as a study model in biological sciences in the latest decades. However, other species of fish also show to be promising as alternatives of use as animal model. This study used Guppy (Poecilia reticulata) as model for the research in behavior through different experimental approaches. In study I, guppies were exposed and re-exposed in different shifts (morning, afternoon, evening and night) in the open field test and light-dark preference. The found results show that in both tests, male and female present behavioral differences, being sensitive to re-exposure, with capacity of learning e controlling of the circadian cycle. In study II, was developed a plus-maze with ramp where it was verified the sensibility of the specie to the apparatus, the profile of response upon re-exposure and the drug effect. The results revealed sensibility to the apparatus with a height of water column at 8 cm and 5 minutes of session and differences between sex and learning by habituation along the re-expositions. The pharmacological study indicates that in this apparatus, the specie is sensitive to anxiolytic and anxiogenic drugs. Finally, we can conclude that Guppy presents similar behavior and drug responses compatible with the data described to the Zebrafish. These similarities reinforce the use of fishes as an alternative to the use of mammals in animal experimentation.

Les ganglions de la base, un ensemble de noyaux sous-corticaux interconnectés, sont impliqués dans l'élaboration, le contrôle et la mémorisation de comportements cognitivo-moteurs. L'une des principales structures de sortie de ce réseau, la substance noire réticulée (SNr), intègre les différentes informations neuronales puis les transmet au cortex via un relais thalamique. Cependant, cette transmission nécessite une régulation fine de l'activité neuronale de la SNr car celle-ci exerce une inhibition constante de ces structures cibles en raison de son activité GABAergique spontanée. Parmi les acteurs de cette régulation, le glutamate et le GABA sont à l'origine d'un équilibre fin entre excitation et inhibition des neurones nigraux. De nombreuses études se sont intéressées aux mécanismes de régulation de l'activité neuronale de la SNr mais, paradoxalement, aucune ne s'est intéressée au rôle des astrocytes. L'objet de ce travail de thèse a donc été d'étudier les relations entre neurones et astrocytes au sein de la SNr, afin de définir une potentielle implication des astrocytes dans la régulation de l'activité neuronale de cette structure. Nous avons étudié les excitabilités calciques des astrocytes et électriques des neurones grâce aux techniques d'imagerie calcique et de patch-clamp, dans un modèle de tranche parasagittale de cerveau de rat préservant les connexions subthalamo-nigrales et pallido-nigrales. Nous avons ainsi montré que les astrocytes nigraux possèdent une activité calcique spontanée, à la fois autonome et dépendante des libérations toniques de glutamate et de GABA. D'autre part, nous avons mis en évidence que l'activité de ces cellules est modulée par la stimulation à haute fréquence du noyau sous-thalamique. Nous avons montré qu'en retour, ces activités calciques spontanées astrocytaires sont impliquées dans la régulation de la fréquence de décharge des neurones de la SNr. Enfin, nous avons mis en évidence que la recapture astrocytaire du glutamate, et probablement du GABA, intervient également dans la régulation de l'activité de décharge neuronale nigrale. En conclusion, ce travail met en évidence une communication bidirectionnelle entre les neurones et les astrocytes de la SNr. Cette communication semble jouer un rôle important dans la régulation de l'activité de cette structure. / Basal ganglia, a set of interconnected nuclei, are implicated in the elaboration, control and memorization of cognitive-motor behaviors. One of the main output structure of this network, the substantia nigra pars reticulata (SNr), integrates and conveys neuronal information to cortical areas via a thalamic relay. However, this transmission requires an accurate regulation of the SNr neuronal activity since this structure inhibits its targets due to its spontaneous GABAergic activity. Among the different actors of this regulation, glutamate and GABA provide a tight balance between excitation and inhibition of the SNr neuronal activity. Several studies have explored the different mechanisms involved in this regulation but paradoxically, none concerned the astrocyte functions. In this work, our aim was to study astrocyte-neuron relations in order to define a potential astrocyte implication in the regulation of the neuronal activity in the SNr. We studied calcium and electrical activities of astrocytes and neurons using calcium imaging and patch-clamp techniques in parasagittal rat brain slices, conserving subthalamo- and pallido-nigral projections. We showed that astrocytes in the SNr displayed spontaneous calcium activities, both dependent and independent of glutamatergic and GABAergic tonic neuronal transmissions. Moreover, we showed that astrocytes calcium activities were regulated by the subthalamic nucleus high frequency stimulation. Our results revealed that, in turn, astrocytes calcium activities were involved in the regulation of the neuronal firing rate. Finally, we showed that astrocyte glutamatergic, and maybe GABAergic, reuptake was involved in the regulation of the neuronal firing rate. To conclude, this study revealed a bidirectional communication between astrocytes and neurons in the SNr. This communication seems to be important in the regulation of the activity in this structure.

Made available in DSpace on 2016-06-02T19:29:38Z (GMT). No. of bitstreams: 1 3822.pdf: 3918575 bytes, checksum: d646b2b0a3b661d4dfa2d3c8a1ee8a69 (MD5) Previous issue date: 2011-03-28 / Universidade Federal de Sao Carlos / The Alto Jacaré-Guaçu Basin was evaluated regarding limnological and ecotoxicological aspects in four water and sediment sampling campaigns at 14 points distributed in the area. Toxicity tests with water, in the laboratory and in situ, showed chronic toxicity, with effects on the survival, growth and gill morphology of juveniles of both fish species examined (Danio rerio and Poecilia reticulata). The lowest survival percentages in the laboratory assays were observed for point L, and in the field tests for point J1. The analysis of the gills revealed first-stage alterations in the specimens of both species exposed at all the sample points, and there were second-stage alterations (aneurism and vascular congestion) at various points. The high concentrations of nutrients and metals in the basin s water and sediments reveal a system impacted by human activities in the surrounding region. The analyses of metals quantified Cd, Cr, Cu, Mn and Fe levels in the water above those permitted by the Brazilian National Environmental Council (CONAMA 357/05), and of Cd in the sediments above the threshold levels permitted both by national (CONAMA 344/04) and international regulations (CCME, 2003). Simple synthetic sediments were prepared composed of sand, clay and organic matter (obtained from the aquatic macrophyte Scirpus cubensis, at concentrations of 0, 0.25, 0.5 and 1%) to assess the toxicity of Cd. The synthetic sediments were viable for the growth and survival of Ceriodaphnia silvestrii, Chironomus xanthus and Danio rerio, and were suitable for use in spiked experiments. The evaluation of the acute toxicity of the sediments fortified with Cd indicated mean 48-h;EC50 for C. silvestrii of 3.09mg.Kg-1 in the sediments with 0% OM and 2.98mg.Kg-1 in the sediments with 1% OM. The mean 96-h;LC50 for C. xanthus was 18.67 mg.Kg-1 in the sediments with 0% OM and nearly four times lower in those with 1% OM (4.93 mg.Kg-1). A similar pattern was noted for D. rerio, a higher 48-h;LC50 in the sediments with 0% OM (114.88 mg.Kg-1) than in the sediments with 1% OM (75.31 mg.Kg-1). In subsequent experiments, the sediments were also spiked with 0.6mg.Kg-1 of Cd, a concentration considered safe for protection of aquatic life according to the applicable national and international regulations. In these tests, the survival of C. silvestrii was impaired in the Cd spiked sediments, so it was not possible to assess the effects on reproduction. There were also effects on the survival, biomass, mentum morphology and Cd concentration in the tissues of C. xanthus. For D. rerio, the survival and growth were compromised in the sediments with higher percentage of organic matter. Besides this, the organisms showed impaired gill morphology and higher Cd concentration in tissue in function of exposure to sediments spiked with the metal. Based on the results, it can be concluded that C. silvestrii was the most sensitive species to the sediments fortified with Cd, followed by C. xanthus and D. rerio, the least sensitive. These effects on the iv organisms at concentrations considered safe for the biota suggest more conservative thresholds of this metal should be established for tropical aquatic habitats. / A bacia do Alto Jacaré-Guaçu foi avaliada quanto aos aspectos limnológicos e ecotoxicológicos em quatro campanhas de amostragem de água e sedimento em 14 pontos distribuídos na área. Testes de toxicidade com água, tanto em laboratório como in situ, apresentaram toxicidade crônica com efeitos na sobrevivência, crescimento e morfologia de brânquias de juvenis de ambas as espécies de peixes avaliadas (Danio rerio e Poecilia reticulata). As menores porcentagens de sobrevivência nos testes em laboratório foram observadas no ponto L e, nos testes in situ, em J1. A análise das brânquias mostrou alterações de primeiro estágio nos organismos expostos em todos os pontos avaliados, e em diversos pontos foram verificadas alterações de segundo estágio (aneurisma e congestão vascular). As elevadas concentrações de nutrientes e metais na água e sedimentos da Bacia do Alto Jacaré-Guaçu revelam um sistema impactado pelas atividades humanas que se desenvolvem em seu entorno. As análises de metais quantificaram Cd, Cr, Cu, Mn e Fe acima da CONAMA 357/05 na água e Cd acima dos valores estabelecidos pela legislação nacional (CONAMA 344/04) e internacional (CCME, 2003) nos sedimentos. Foram desenvolvidos sedimentos sintéticos simples compostos de areia, argila e matéria orgânica (obtida a partir da macrófita aquática Scirpus cubensis, nas concentrações de 0; 0,25; 0,5 e 1%) para avaliar a toxicidade do Cd. Os sedimentos sintéticos se apresentaram viáveis para o crescimento e sobrevivência de Ceriodaphnia silvestrii, Chironomus xanthus e Danio rerio, sendo adequados para uso em experimentos de fortificação. A avaliação da toxicidade aguda dos sedimentos fortificados com Cd, indicaram CE50;48h média para C. silvestrii de 3,09mg.Kg-1 nos sedimentos sem MO e 2,98mg.Kg-1 nos sedimentos com 1% de MO. A CL50;96h média para C. xanthus foi de 18,67mg.Kg-1 nos sedimentos sem MO e, cerca de 4 vezes menor nos sedimentos com 1% de MO, 4,93mg.Kg-1. Para D. rerio também foi notada tendência semelhante, CL50;48h maior nos sedimentos sem MO, 114,88mg.Kg-1 e menor nos sedimentos com 1% de MO, 75,31mg.Kg-1. Os sedimentos também foram fortificados com 0,6mg.Kg-1 de Cd, concentração considerada segura para a proteção da vida aquática segundo legislações nacionais e internacionais, em experimentos posteriores. C. silvestrii teve a sobrevivência comprometida nos sedimentos fortificados com Cd, de modo que não foi possível avaliar os efeitos na reprodução. C. xanthus também apresentou efeitos na sobrevivência, biomassa, morfologia de mento e concentração de Cd nos tecidos. D. rerio teve a sobrevivência e crescimento comprometidos nos sedimentos com maior porcentagem de matéria orgânica. Além disso, os organismos apresentaram a morfologia de brânquias comprometida e concentração de Cd em ii seus tecidos elevada em função da exposição aos sedimentos fortificados com o metal. Com base nos resultados foi possível concluir que C. silvestrii foi a espécie mais sensível aos sedimentos fortificados com Cd, seguido de C. xanthus e, a menos sensível, D. rerio. Tais efeitos observados nos organismos em concentrações consideradas seguras para a biota sugerem a avaliação mais cuidadosa dos valores para espécies e climas tropicais.

No description available.

Citrus es el género de la subfamilia Aurantioideae de mayor importancia económica. Su origen es la región sureste de Asia, en un área que incluye China, India y la península de Indochina y los archipiélagos de los alrededores. Aunque se han realizado múltiples estudios, la taxonomía del género Citrus aun no está bien definida, debido al alto nivel de diversidad morfológica encontrado en este grupo, la compatibilidad sexual entre sus especies y la apomixis de muchos genotipos. En la presente tesis doctoral se ha estudiado una amplia diversidad del género Citrus, especies relacionadas y otros taxones de la subfamilia Aurantioideae, para poder aclarar su organización y filogenia mediante el empleo de diferentes tipos de marcadores moleculares y métodos de genotipado. Más concretamente, el germoplasma de mandarino juega un papel muy importante en la mejora de variedades y patrones, pero su organización genética no está bien definida. Por lo tanto, se ha realizado un análisis en profundidad de su diversidad y organización genética. El desarrollo de marcadores moleculares de Inserción-Deleción (indel), por primera vez en cítricos, ha permitido demostrar su utilidad para estudios de diversidad y filogenia en el género Citrus. En combinación con los marcadores de tipo microsatélite (SSR), se ha cuantificado la contribución de los tres principales taxones de cítricos (C. reticulata, C. maxima and C. medica) a los genomas de las especies secundarias y cultivares modernos. También se ha definido su estructura genética a partir de los datos obtenidos en la secuenciación de 27 fragmentos de genes nucleares relacionados con la biosíntesis de compuestos que determinan la calidad de los cítricos y genes relacionados con la respuesta de la planta a estreses abióticos. El análisis de la filogenia nuclear ha permitido determinar la relación existente entre la especie C. reticulata y Fortunella, que se diferencian claramente del grupo formado por las otras dos principales especies de cítricos (C. maxima y C. medica). Este resultado está en concordancia con el origen geográfico de las especies estudiadas. A partir de este estudio, se han desarrollado marcadores moleculares de tipo SNP con un alto valor filogenético, que han sido transferidos a géneros relacionados de los cítricos. Estos marcadores han dado un resultado muy positivo en el género Citrus y serán de gran utilidad para el establecimiento de la huella genética del germoplasma en un nivel de diversidad más amplio. Se ha estudiado la organización genética dentro del germoplasma mandarino (198 genotipos de tipo mandarino pertenecientes a dos colecciones, INRA-CIRAD e IVIA), así como la introgresión de otros genomas mediante el uso de 50 y 24 marcadores de tipo SSR y indel, respectivamente, además de cuatro marcadores InDel mitocondrial (ADNmt). Se ha observado que muchos genotipos, que se creía que eran mandarinos puros, presentan introgresión de otros genomas ancestrales. Dentro del germoplasma de mandarino, se han identificado a nivel nuclear cinco grupos parentales, a partir de los cuales se originaron muchos genotipos, dando lugar a estructuras hibridas complejas. Se ha observado incluso, genotipos con un origen maternal no mandarino, determinado por los marcadores de ADNmt. La presente tesis doctoral ha aportado nueva información sobre las relaciones filogenéticas entre las especies del género Citrus, géneros cercanos, así como de las especies secundarias. Además, se han desarrollado nuevos marcadores moleculares que se complementan entre sí. Se ha establecido una nueva organización genética del germoplasma mandarino y se han caracterizado adecuadamente las dos colecciones de cítricos en estudio. Por lo tanto, todas estas contribuciones, ayudarán a los programas de mejora para la obtención de nuevas variedades de cítricos de alta calidad y permitirán optimizar la conservación y uso de los recursos genéticos existentes, así como su caracterización genética y fenotípica. / García Lor, A. (2013). Organización de la diversidad genética de los cítricos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/31518

Le récepteur aux cannabinoïdes de type 1 (CB1) est un récepteur couplé aux protéines G, abondamment exprimé dans le cerveau et régulant plusieurs processus physiologiques. Cependant, les mécanismes cellulaires par lesquels les CB1 régulent ces processus n’ont été que peu analysés. Bien que les CB1 localisés dans les membranes plasmiques sont connus pour induire la transduction de signal; une partie de ces récepteurs sont aussi fonctionnels au niveau des mitochondries (mtCB1), où leur stimulation réduit la respiration mitochondriale. L’objectif de cette thèse fut d’évaluer l’impact de l’activation des récepteurs mtCB1 du cerveau sur les effets connus des cannabinoïdes. Afin de distinguer la fonction des mtCB1 de celle des autres populations de récepteurs, nous avons développé des outils basés sur la signalisation induite par les mtCB1. Dans les mitochondries isolées de cerveau, l’activation des protéines Gαi/o, dépendante des mtCB1 diminue l’activité de l’adénylyl cyclase soluble (sAC). L'inhibition locale de l’activité de sAC prévient l’amnésie, la catalepsie et partiellement l’hypolocomotion induite par les cannabinoïdes. De plus, nous avons généré une protéine fonctionnelle mutante CB1 (DN22-CB1) dépourvue des 22 premiers acides aminés des CB1 ainsi que de sa localisation mitochondriale. Contrairement aux CB1, l'activation des DN22-CB1 n’affecte pas l'activité mitochondriale. Enfin, l’expression des DN22-CB1 dans l’hippocampe bloque à la fois la diminution de la transmission synaptique et l’amnésie induites par les cannabinoïdes. Ces travaux démontrent l’implication des mtCB1 dans certains effets des cannabinoïdes et le rôle clé des processus bioénergétiques contrôlant les fonctions cérébrales. / Type-1 cannabinoid receptor CB1 is a G protein-coupled receptor (GPCR), widely expressed in the brain, which regulates numerous physiological processes. However, the cellular mechanisms of CB1-mediated control of these functions are poorly understood. Although CB1 are known to signal at the plasma membrane, a portion of these receptors are also present in mitochondria (mtCB1), where mtCB1 activation decreases mitochondrial activity. The goal of this thesis was to dissect the impact of brain mtCB1 signaling in known behavioral effects induced by cannabinoids. To distinguish the functions of mtCB1 from other receptor pools, we developed tools based on the characterization of the intra-mitochondrial molecular cascade induced by mtCB1 receptors. In isolated brain mitochondria, we found that intra-mitochondrial decrease of soluble-adenylyl cyclase (sAC) activity links mtCB1- dependent activation of Gαi/o proteins to decrease cellular respiration. Local brain inhibition of sAC activity blocks cannabinoid-induced amnesia, catalepsy and contributes to the hypolocomotor effect of cannabinoids. In addition, we generated a functional mutant CB1 protein (DN22-CB1) lacking the first 22 amino acid of CB1 and its mitochondrial localization. Differently from CB1, activation of DN22-CB1 does not affect mitochondrial activity. Hippocampal in vivo expression of DN22-CB1 abolished both cannabinoid-induced impairment of synaptic transmission and amnesia in mice. Together, these studies couple mitochondrial activity to behavioral performances. The involvement of mtCB1 in the effects of cannabinoids on memory and motor control highlights the key role of bioenergetic processes as regulators of brain functions.