In addition to previously identified roles for RNA, several new ways in which RNA serves as a regulator of gene expression have recently been described. RNA molecules are involved in both transcriptional and post-transcriptional forms of regulation, sometimes heritably affecting gene activity. Whereas most previously characterized regulatory roles of RNA involve downregulation, I describe a role for maternal transcripts of a gene in promoting zygotic activity of that gene, which I term the licensing of genetic activity. This regulation occurs in the germ line, a tissue notable for its abundance of genetic surveillance mechanisms. The maternal-effect regulation described here was identified using alleles of a sex-determining gene in Caenorhabditis elegans called fem-1. Females homozygous for fem-1 deletions produce heterozygous offspring that exhibit germ-line feminization and have reduced fem-1 activity and transcript accumulation. This phenotype can be rescued by injecting fem-1 RNA into the maternal germ line. The reduction in activity of the zygotic fem-1 locus is heritable, suggesting that the gene is becoming epigenetically silenced. Thus the maternal fem-1 RNA licenses the activity of the zygotic locus by preventing its silencing. By restricting germ-line activity to genes that were expressed in the germ line of the previous generation, this process may contribute to protecting the identity and integrity of the germ line. I performed an RNAi screen of candidate genes to ask whether they are required for maternal-effect silencing or licensing. Several enhancers and suppressors of germ-line feminization in the descendants of fem-1 deficiency homozygotes were identified. Chromatin regulation may be involved, and small-RNA pathways are important for both the silencing and licensing components of fem-1 regulation. Based on my characterization of this phenomenon, I proposed models of how maternal-effect regulation of fem-1 may be mediated. To test predictions of certain models, I examined whether specific characteristics of fem-1 make it susceptible to this silencing. Results of these experiments limit the possible models of maternal-effect regulation and suggest directions for future investigation.

In addition to previously identified roles for RNA, several new ways in which RNA serves as a regulator of gene expression have recently been described. RNA molecules are involved in both transcriptional and post-transcriptional forms of regulation, sometimes heritably affecting gene activity. Whereas most previously characterized regulatory roles of RNA involve downregulation, I describe a role for maternal transcripts of a gene in promoting zygotic activity of that gene, which I term the licensing of genetic activity. This regulation occurs in the germ line, a tissue notable for its abundance of genetic surveillance mechanisms. The maternal-effect regulation described here was identified using alleles of a sex-determining gene in Caenorhabditis elegans called fem-1. Females homozygous for fem-1 deletions produce heterozygous offspring that exhibit germ-line feminization and have reduced fem-1 activity and transcript accumulation. This phenotype can be rescued by injecting fem-1 RNA into the maternal germ line. The reduction in activity of the zygotic fem-1 locus is heritable, suggesting that the gene is becoming epigenetically silenced. Thus the maternal fem-1 RNA licenses the activity of the zygotic locus by preventing its silencing. By restricting germ-line activity to genes that were expressed in the germ line of the previous generation, this process may contribute to protecting the identity and integrity of the germ line. I performed an RNAi screen of candidate genes to ask whether they are required for maternal-effect silencing or licensing. Several enhancers and suppressors of germ-line feminization in the descendants of fem-1 deficiency homozygotes were identified. Chromatin regulation may be involved, and small-RNA pathways are important for both the silencing and licensing components of fem-1 regulation. Based on my characterization of this phenomenon, I proposed models of how maternal-effect regulation of fem-1 may be mediated. To test predictions of certain models, I examined whether specific characteristics of fem-1 make it susceptible to this silencing. Results of these experiments limit the possible models of maternal-effect regulation and suggest directions for future investigation.

No description available.

A genetic screen for dominant, temperature-sensitive, maternal-effect embryonic lethal mutations identified mel-43(sb41), a gene required for early embryonic viability (Mitenko et al., 1997). Linkage mapping placed mel-43 within a small region on chromosome IV. Genetic analyses suggested that mel-43(sb41) was a neomorphic mutation. While refining the genetic position of the mel-43 gene, data suggested that the genetic position of mel-43 was inconsistent with the published location. In light of this new location, previous conclusions regarding the genetic behaviour of mel-43(sb41) were re-examined. Deficiency analysis suggests that mel-43(sb41) is a haploinsufficient loss-of-function mutation. mel-43(sb41) embryos are significantly delayed in meiosis II independent of cyclin B1 degradation. Consequently, embryos fail to produce meiosis II polar bodies and do not establish proper polarity. Although the function of mel-43 remains unknown, the persistent meiotic spindle suggests that mel-43 acts upstream of the microtubule rearrangements necessary to promote the metaphase II to anaphase II transition. / Molecular Biology and Genetics

Le transfert intergénérationnel de l'immunité est un phénomène dont l'étude remonte à une dizaine d'année. Chez les vertébrés, il prend la forme d'un transfert d'anticorps de la mère aux juvéniles. Pourtant, les processus écologiques et évolutifs du transfert sur plusieurs générations et la persistance des anticorps maternels chez les juvéniles n'ont pas bien étudiés. Ce travail de thèse porte sur l'étude des mécanismes du transfert d'anticorps maternel sur plusieurs générations consécutives et leurs conséquences sur la réponse immunitaire et sur la croissance des juvéniles. En utilisant des approches expérimentales, j'ai pu dans un premier temps montré que le transfert d'anticorps maternel était affecté par la disponibilité en nourriture dans l'environnement. De plus, nos résultats suggèrent que la persistance d'anticorps maternel dépend du niveau initial d'anticorps maternels présents chez le juvénile dans les premiers jours de vie. Dans un second temps, nous avons étudié la réponse immunitaire et la croissance des poussins en tentant de mettre en évidence comment les anticorps maternels influençaient le compromis entre ces deux paramètres physiologiques. Enfin, nos résultats suggèrent que les anticorps maternels pourraient être le messager intergénérationel d'un effet épigénétique sur la réponse immunitaire. En effet, une réponse immunitairre des grand-mères induit par l'exposition à un parasite a été trouvé stimuler la réponse immunitaire des petits-enfants contre ce même parasite. Mes travaux de thèses soulignent une fois de plus, l'importance du transfert de l'immunité dans l'écologie et l'évolution des interactions hôte-parasite. / Intergenerational transfer of immunity is a phenomenon which started to be studied 10 years ago. In vertebrates, it takes the form of a transfer of antibodies from mothers to juveniles. However, the ecological and evolutionary processes of transfer across several generations and the persistence of maternals antibodies in chicks remain poorly understood. This thesis focuses on the study of the mechanisms of maternal antibodies transfer over several generations and on how they impact the immune response and growth in juveniles. By using experimental approaches, I first show that the transfer of maternal antibody was affected by food availability in the environment. In addition, our results suggest that the persistence of maternal antibodies depends on the initial level of maternal antibodies present in the nestlings in the first days of life. Secondly, we studied the immune response and growth of juvenile by investigating how maternal antibodies may affect the resolution of the trade-off between these two physiological parameters. Finally, our results suggest that maternal antibodies would be a messenger of an epigenetic effect on immunty. Indeed, induced immune response in grand-mothers towards a pathogen has been found to stimulate the immune response of grand-children. My thesis emphasizes the importance of maternal transfer of immunity in the ecology and evolution of host-parasite.

Made available in DSpace on 2016-08-10T10:44:43Z (GMT). No. of bitstreams: 1 BRUNO FRAUZINO RIBEIRO CAMILO.pdf: 1862817 bytes, checksum: ba7c8b48dadf912fb820375d1be59050 (MD5) Previous issue date: 2014-12-15 / The objective was to study the effects on the ordering of the animals by their genetic value of the inclusion of direct maternal additive genetic covariance models of prediction of breeding values for weaning weight in beef cattle using simulated data. The data were simulated considering three different values of maternal direct additive genetic correlation (-0.25, -0.50 and +0.25) and three reasons direct additive variance and maternal additive (1: 1, 1: 2 and 2: 1), making a total of nine scenarios. In each scenario, the breeding values were predicted including whether or not the covariance in the model. The values of the Spearman correlation between animal clinics, considering the predicted genetic value, the scenarios of direct maternal additive genetic correlation -0.50 in the ratios of variances 1: 1 and 2: 1 were equal to 0.84 and 0, 85, respectively; -0.25 scenario the correlation variance in the ratios 1: 1, 1: 2 and 2: 1 were 0.96, 0.95 and 0.97 respectively, and the positive correlation scenario in +0.25 Variance ratios 1: 1, 1: 2 and 2: 1 with values of 0.92, 0.91 and 0.96 respectively. The values were high and significant, suggesting greater consistency between the animal merit orders. The average value for the Spearman correlation estimated in scenario - 0.50 1: 2 was equal to 0.46 indicating lesser proximity between merit orders of individuals in this scenario. The results indicate include the covariance in prediction models for the classification of animals are closer to real. / Objetivou-se estudar os efeitos, sobre a ordenação dos animais por seu valor genético, da inclusão da covariância genética aditiva direta maternal nos modelos de predição dos valores genéticos de peso a desmama de bovinos de corte utilizando dados simulados. Os dados foram simulados considerando-se três valores diferentes de correlação genética aditiva direta maternal (-0,25, -0,50 e +0,25) e três razões de variância aditiva direta e aditiva maternal (1:1, 1:2 e 2:1), perfazendo um total de nove cenários. Em cada cenário, os valores genéticos foram preditos incluindo-se ou não a covariância no modelo. Os valores da correlação de Spearman entre os postos dos animais, considerando o valor genético predito, nos cenários de correlação genética aditiva direta maternal -0,50 nas razões de variâncias 1:1 e 2:1 foram iguais a 0,84 e 0,85, respectivamente; no cenário de correlação -0,25 nas razões de variâncias 1:1, 1:2, e 2:1 foram iguais a 0,96, 0,95 e 0,97 respectivamente e no cenário de correlação positiva +0,25 nas razões de variâncias 1:1, 1:2 e 2:1 com valores de 0,92, 0,91 e 0,96 respectivamente. Os valores foram altos e significativos, o que sugere maior concordância entre as ordens de mérito dos animais. O valor médio para a correlação de Spearman estimados no cenário -0,50 1:2 foi igual a 0,46 indicando menor proximidade entre as ordens de mérito dos indivíduos neste cenário. Os resultados indicam incluir a covariância nos modelos de predição para que as classificações dos animais estejam mais próximas do real.

Les moustiques jouent un rôle essentiel dans la dynamique de transmission du paludisme. Plusieurs traits d’histoire de vie de ces insectes hématophages vont, en effet, intervenir dans le calcul du taux reproductif de base du parasite. Parmi eux, la probabilité et l’intensité d’infection des vecteurs, suite à la prise d’un repas de sang infectieux, sont des facteurs primordiaux pour la transmission de Plasmodium. Pourtant, au sein d'une même population, une réelle hétérogénéité d’infection des moustiques a été observée: alors que certain ne sont pas infectés, que d'autres le sont faiblement, une partie des vecteurs vont présenter des taux d'infection extrêmement forts. Identifier les sources de variations responsables de cette hétérogénéité est indispensable pour comprendre la dynamique de transmission de Plasmodium. Certains paramètres d’origine génétique et environnementale, tel que la température et la nutrition, ont déjà clairement été identifiés comme ayant un impact sur le taux d’infection des moustiques. Néanmoins, de nombreux facteurs restent encore à explorer. En utilisant un système expérimental composé du parasite de la malaria aviaire Plasmodium relictum, de son vecteur naturel le moustique Culex pipiens et de l’un de ses hôtes vertébrés Serinus canaria, nous nous sommes intéressés aux effets de certains de ces paramètres, peu pris en compte jusqu’à présent, sur l’infection des moustiques. Nous avons notamment pu mettre en évidence des conséquences importante de l’infection parentale, de l’âge des vecteurs, ainsi que de leur fond génétique. Parallèlement, nous nous sommes intéressés à un facteur souvent laissé de côté, l’effet « hôte vertébré » sur l’infection des moustiques. Dans cette dernière partie, on a pu observer que la dynamique d’infection du parasite au sein de l’hôte influence de manière importante le taux de transmission de Plasmodium. Les sources d’hétérogénéité d’infections des moustiques sont donc multiples et l’ensemble des membres de cette association tripartite hôte/parasite/vecteur vont pouvoir influencer de manière importante la dynamique de transmission du parasite. / Mosquitoes play a key role in the dynamics of malaria transmission. Indeed, several life history traits of these bloodsucking insects are closely associated with the basic reproductive rate of the malaria parasite. One of the most important parameters for the transmission of Plasmodium is the rate and intensity of the mosquito infection. However, within a single population, a great heterogeneity of mosquito infection levels is often observed: some mosquitoes are not infected, others are only weakly infected, and yet others have extremely high infection rates. Identifying the sources of variation responsible for this heterogeneity is essential in order to understand the transmission dynamics of Plasmodium. Certain genetic and environmental (temperature, nutrition) parameters have already been identified as having an impact on mosquito infection rates. Nevertheless, many factors remain to be explored. Using an experimental system composed of the avian malaria parasite Plasmodium relictum, its natural vector Culex pipiens and one of its vertebrate host Serinus canaria, we have investigated the effect of several seldom investigated parameters on the infection of mosquitoes. We observed surprising effects of the effect of parental infection, mosquito age and genetic background. In parallel, we also studied a rarely investigated parameter: the impact of the vertebrate host on the mosquito infection rate. We showed that parasite infection dynamics within the host, at short but also long time scales, significantly influence the transmission of Plasmodium to the vector. The sources of heterogeneity of mosquito infection are therefore multiple and all members of this tripartite partnership (i.e. host / parasite / vector) are able to significantly influence the transmission dynamics of the parasite.

<p>Haldane’s rule states “when in the F1 offspring of two different animal races one sex is absent, rare, or sterile, that sex is the heterozygous sex”. Many studies regarding hybrid male sterility and in-viability have been done in order to better understand the process of speciation in males. Yet the study of speciation in hybrid females has been largely ignored. In our study we re-examined the extent of fertility in F1 reciprocal females of hybridization between <em>Drosophila simulans and D. mauritiana</em> species<em>.</em> Hybridization between these species produces fertile females and sterile males. Our goal was to address the following questions: 1. Are F1 hybrid females fully fertile? 2. Are there any maternal effects observed in reciprocal female hybrids? 3. Are there significant differences in ovariole numbers between the reciprocal hybrids? and 4. What is the state of the hybrid ovaries as a function of age? In order to answer these questions we looked at the level of oviposition and egg hatchability and the differences in ovariole numbers in pure species and F1 females. Our results indicated that the reciprocal hybrid females are not only fully fertile but they also showed heterosis. The heterosis observed in the hybrids can be attributed in part to the presence of maternal effects. The reciprocal hybrids also showed differences in ovariole number compared to each other and to the parental species. Our findings signify the importance of maternal effects as a potentially powerful mechanism for moderating the rates of evolution of speciation in hybrid females.</p> / Master of Science (MSc)

The aim of this thesis project was to elucidate patterns and processes associated with the biotic interactions in a natural plant–herbivore–parasitoid food web characterized by spatial and temporal heterogeneity with regard to species composition. The system examined is based on island populations of the perennial herb Meadowsweet (Filipendula ulmaria, Rosaceae), located in the Skeppsvik Archipelago. The area is subject to isostatic rebound, amounting to 0.85 cm per year; this makes it possible to calculate the age of the rising islands. Meadowsweet colonizes new islands when they are about 100 years old. Meadowsweet is consumed by two major herbivores in the study area: Galerucella tenella and Altica engstroemi (Coleoptera: Chrysomelidae). Both herbivores overwinter in the topsoil and successful colonization occurs when the islands reach a height that prevents the beetles from being removed or killed as a result of wave wash during the winter. I found that both herbivores significantly reduced individual plant fitness and population growth rate. A “cafeteria experiment” with Galerucella showed that this beetle discriminated between plants from different islands, avoiding plants from old islands which contained high concentrations of putative defence compounds, while readily accepting plants from younger islands which contained lower concentrations of these chemicals. Further, the plant species exhibited a trade-off between growth and production of the putative defence compounds. Taken together, these results were interpreted as providing evidence of herbivore-driven evolution of resistance in Meadowsweet. Further, laboratory studies suggested that Galerucella gradually includes a less preferred host plant (Rubus arcticus, Rosaceae) in its diet as Meadowsweet resistance increases. This implies that Galerucella drives its own host-breadth enlargement by selectively inducing a ‘rent rise’ in the original host, Meadowsweet. In a number of field studies I showed that the oligophagous parasitoid Asecodes mento (Hymenoptera: Eulophidae) has a strong positive effect on Meadowsweet seed set by removing large numbers of G. tenella larvae. This top-down effect is, however, altered by the presence of a close relative of G. tenella, namely G. calmariensis, which is monophagous on Purple loosestrife (Lythrum salicaria, Lythraceae). G. tenella experiences associational susceptibility when coexisting with G. calmariensis since the latter supports a higher and more fit pool of shared parasitoids and because Meadowsweet attracts a higher proportion of the shared parasitoid females than Purple loosestrife. This implies that G. tenella densities are very low in coexisting populations and that Meadowsweet experiences associational resistance and produces more seeds when co-occurring with Purple loosestrife. Thus, selection for increased resistance in Meadowsweet is likely to be relaxed in populations mixed with Purple loosestrife. I conclude that the evolution of plant resistance is likely to depend on the length of time and intensity of selection. When Meadowsweet colonizes new islands it experiences a period of enemy-free space; followed by a midlife and ageing with selection by herbivores. The intensity of this selection does, however, depend on the presence of additional plant and herbivore species.

Maximiser le nombre de copies de gènes transmises aux générations suivantes implique une série de compromis. Chez les espèces à reproduction coopérative, des individus ne se reproduisent pas mais participent aux soins des jeunes d'autres individus reproducteurs. Ces assistants sont particulièrement intéressants dans le contexte des traits d'histoire de vie car ils forment un environnement prédictible favorable pour la reproduction, et leur présence peut aussi influencer les compromis évolutifs chez les reproducteurs. Un compromis évolutif majeur mais sous-étudié dans le cadre de la reproduction coopérative est l'allocation maternelle notamment via des effets maternels qui sont des modifications épigénétiques du phénotype de la descendance. Nous avons étudié l'existence d'effets maternels associés à la présence d'assistants et leurs possibles conséquences sur les femelles et leurs descendants chez un oiseau colonial et coopératif du sud de l'Afrique, le Républicain social Philetairus socius. Nos résultats montrent que les femelles pondent des œufs plus légers en présence d'assistants et que ces œufs sont moins concentrés en corticostérone et testostérone. Nos résultats montent aussi une plus grande probabilité de survie pour les femelles se reproduisant en groupe pouvant être en partie due à leur plus faible investissement dans les œufs. De plus, l'étude de la température dans les nids en fonction de la taille des groupes a permis de suggérer d'autres bénéfices pour les parents et assistants, en particulier via une réduction des coûts de thermorégulation qui pourrait aussi permettre de garder de l'énergie pour la survie. Pour comprendre les conséquences de la présence d'assistants et de l'allocation différentielle pour les poussins, une expérience d'adoption croisée a été réalisée. Elle a révélé que les œufs pondus par les femelles avec plus d'assistants produisent des poussins qui quémandent moins, montrant que des effets maternels pourraient influencer le comportement des poussins. Enfin nous avons étudié la survie des poussins après l'envol à l'aide d'analyses de captures recaptures et avons trouvé de manière surprenante que les poussins à l'envol ont une probabilité de survie plus faible lorsqu'ils sont élevés présence d'assistants. Ces résultats dans leur ensemble démontrent l'importance d'étudier les effets maternels chez les espèces coopératives et ouvrent de nombreuses perspectives de recherche sur les conflits familiaux et de compromis évolutifs associés à la présence d'assistants. / Maximizing of the number copies of genes that are transmitted to the next generations involves a series of tradeoffs. In cooperatively breeding species some sexually mature individuals do not breed but instead help other individuals to raise their offspring. These helpers are particularly interesting in a life history context as they create a predictably favorable breeding environment and their presence can thus influence evolutionary trade-offs. A major evolutionary trade-off that is often neglected in studies on cooperative breeding is maternal allocation, notably through maternal effects that are epigenetic modifications of offspring phenotype. Here we investigate whether there are maternal effects induced by the presence of helpers and their possible consequences on females and their offspring in a colonial cooperative breeder of southern Africa, the sociable weaver Philetairus socius. Our results show that females lay smaller eggs in the presence of helpers and in addition these eggs have lower corticosterone and testosterone concentrations. Our results also show a higher survival probability of females breeding in groups, which may be partially due to their lower investment in eggs. In addition, a study of roosting chamber temperatures in relation to group size suggests further benefits for parents and helpers, particularly through lower costs of thermoregulation that could also allow energy savings for survival. To start understanding the consequences of helpers presence and differential maternal allocation for offspring we conducted a cross fostering experiment. Our results show that eggs produced by females breeding in larger groups produce chicks that beg at a lower rate, showing that maternal effects may influence chicks' behavior. Finally, we investigated post-fledging survival through capture-recapture analyses and, surprisingly, found that fledglings have a lower survival probability when raised with helpers. Taken together, these results demonstrate the importance of studying maternal effects on cooperative breeders and open several research prospects on family conflicts and life history trade-offs according to the presence of helpers.