The human-abetted introduction of commensal species (i.e. those that opportunistically exploit the anthropogenic environment for food and shelter, e.g. rats, cockroaches etc.) to new areas has occurred throughout history. This has resulted in detrimental ecological changes worldwide but, from a viewpoint of human knowledge, a beneficial corollary of these translocations is that the species in question can be used as proxies to study the movement of the humans who transported them. I reconstruct colonisation histories of three widespread commensal mammalian species in the Western Indian Ocean, the black rat Rattus rattus, house mouse Mus musculus and Asian house shrew Suncus murinus, through phylogeographic studies (the geographic distribution of genetic lineages) of maternally-inherited mitochondrial markers, and zooarchaeological data. The DNA analyses are conducted on samples largely derived from museum specimens collected up to 110 years ago, and from archaeological bones (in the case of rats). I show considerable cryptic diversity in all three species, particularly in mice for which we find a potential major new lineage. Certain lineages within each species predominantly reveal long-distance translocations within the Indian Ocean, but high resolution geographic and genetic clustering is also evident, particularly in Asian house shrews. Phylogeographic structuring of the three species in East Africa and the southern Indian Ocean region (e.g. Madagascar, Reunion, etc.) indicate connections with Arabia, the Middle East, and India in the Islamic period from the first millennium AD, and later European connections during the Age of Exploration. Closer to the origins of the three species (the Indian subcontinent in all cases), range expansions in Eurasia and nearby islands relate to early to mid Holocene human populations, but also with signals of later secondary colonisations. Through ancient DNA studies I found genetic continuity between temporally separated populations of black rats suggesting population persistence, and high levels of diversity in Songo Mnara, a Swahili stonetown in Tanzania. Knowledge of the colonisation history and genetic diversity of an introduced species is essential to understand their resilience in novel landscapes, and to identify pathways of invasion and, by proxy, human trade and exchange networks that facilitated their dispersal. My research contributes significantly to that end for three socially, economically and ecologically important species that are well-established in the Indian Ocean region and beyond.

Neurogenin 2 (Ngn2) is a basic Helix-Loop-Helix (bHLH) transcription factor regulating differentiation and cell cycle exit in the developing brain. By transcriptional upregulation of a cascade of other bHLH factors, neural progenitor cells exit the cell cycle and differentiate towards a neuronal fate. Xenopus laevis Ngn2 (xNgn2) is a short-lived protein, targeted for degradation by the 26S proteasome. I have investigated the stability of Ngn2 mediated by post-translational modifications and structural disorder. Firstly I will describe work focused on ubiquitylation of xNgn2, targeting it for proteasomal degradation. xNgn2 is ubiquitylated on lysines, the recognized site of modification. I will discuss the role of lysines in ubiquitylation and stability of xNgn2. In addition to canonical ubiquitylation on lysines, I describe ubiquitylation of xNgn2 on non-canonical sites, namely its amino-terminal amino group, and cysteine, serine and threonine residues. I show that the ubiquitylation of cysteines in particular exhibits cell cycle dependence and is also observed in mammalian cell lines, resulting in cell cycle-dependent regulation of stability. I will then discuss whether phosphorylation, a regulator of xNgn2 activity, also affects xNgn2 stability. I will provide evidence of cell cycle-dependent phosphorylation of cyclin dependent kinase (cdk) consensus sites affecting the stability of xNgn2. Finally I describe studies on the folding properties of Ngn2 to assess their role in protein stability. xNgn2 associates with DNA and its heterodimeric binding partner xE12 and may interact directly with the cyclin-dependent kinase inhibitor Xic1. I will discuss the role of these interaction partners in xNgn2 stability. xNeuroD, a downstream target of xNgn2, is a related bHLH transcription factor which is stable. Here I describe domain swapping experiments between these two proteins highlighting regions conferring instability on the chimeric protein. Finally I will provide nuclear magnetic resonance (NMR) data looking at the effect of phosphorylation on protein structure in mouse Ngn2 (mNgn2).

The coexistence of humans and rodents lasts from the beginnings of the history of agriculture. Many rodent populations accepted to synanthropic way of life and as commensals accompany human societies until today. In the first study we wanted to find out, how the evolution of non-commensal rodent species, a Cypriot mouse (Mus cypriacus), was influenced by people. This endemic island species evolved on the Cyprus Island without presence of any mammalian competitors or predators. In last 10.000 years humans arrived on the island, bringing several mammalian species, which affected the environment markedly. A black rat (Rattus rattus) is now dominant species there and presumably it is an important competitor for the Cypriot mouse. We supposed that the mice should avoid its odour. Nevertheless the rats odour was preferred by the mouse, probably as an odour of phylogenetically related species. On the contraty, the odour of domestic cat (species, which is also widespread on the island), was avoided. In conclusion, the long-term isolation from mammalian predators did not affect the antipredatory reactions of the Cypriot mouse. Our next study was focuse on changes in behavioural strategies of different populations of mice: the main question was how the commensal way of life affects their exploratory...

Los volcanes son formaciones geológicas que resultan atractivas para los asentamientos humanos debido a la fertilidad de sus suelos y su interés turístico. Pero también son una fuente importante de contaminación natural, debido a los procesos geoquímicos que tienen lugar durante los periodos eruptivos y no eruptivos. Se estima que 44 millones de personas viven a menos de 10 km de un volcán activo y ya se ha demostrado el aumento de incidencia de patologías respiratorias en humanos que habitan estos ambientes. La presente Tesis Doctoral tiene como objetivo valorar los efectos neurotóxicos en el sistema nervioso central de ratones domésticos (Mus musculus) que habitan en ambientes volcánicos activos. La acumulación de mercurio en cerebro y médula espinal de individuos expuestos, la activación de células astrocitarias y microgliales y la producción de citocinas proinflamatorias son los principales resultados de este trabajo de los que se desprende que el sistema nervioso central está afectado por la exposición crónica a estos ambientes.

The coexistence of humans and rodents lasts from the beginnings of the history of agriculture. Many rodent populations accepted to synanthropic way of life and as commensals accompany human societies until today. In the first study we wanted to find out, how the evolution of non-commensal rodent species, a Cypriot mouse (Mus cypriacus), was influenced by people. This endemic island species evolved on the Cyprus Island without presence of any mammalian competitors or predators. In last 10.000 years humans arrived on the island, bringing several mammalian species, which affected the environment markedly. A black rat (Rattus rattus) is now dominant species there and presumably it is an important competitor for the Cypriot mouse. We supposed that the mice should avoid its odour. Nevertheless the rats odour was preferred by the mouse, probably as an odour of phylogenetically related species. On the contraty, the odour of domestic cat (species, which is also widespread on the island), was avoided. In conclusion, the long-term isolation from mammalian predators did not affect the antipredatory reactions of the Cypriot mouse. Our next study was focuse on changes in behavioural strategies of different populations of mice: the main question was how the commensal way of life affects their exploratory...

The cells from the inner cell mass (ICM) of an early embryo have the potential to differentiate into all the different cell types present in an adult organism. Cells from the ICM can be isolated and cultured in vitro, becoming embryonic stem cells (ESCs). ESCs have several properties that make them unique: they are unspecialized, can self-renew indefinitely in culture, and given the appropriate cues can differentiate into cells from all three germ layers (ecto-, meso-, and endoderm), including the germline, both in vivo and in vitro. Induced pluripotent stem cells (iPSCs) can be generated from adult, terminally differentiated somatic cells by transient exogenous expression of four transcription factors (Oct4, Sox2, Klf4, and cMyc; OSKM) present normally in ESCs. It has been shown that iPSCs are equivalent to ESCs in terms of morphology, gene expression, epigenetic signatures, in vitro proliferation capacity, and in vitro and in vivo differentiation potential. However, unlike ESCs, iPSCs can be obtained from a specific individual without the need for embryos. This makes them a promising source of pluripotent cells for regenerative medicine, tissue engineering, drug discovery, and disease modelling; additionally, in livestock species such as the bovine, they also have applications in genetic selection, production of transgenic animals for agricultural and biomedical purposes, and species conservancy. Nevertheless, ESC and iPSC lines that meet all pluripotency criteria have, to date, only been successfully produced in mice, rats, humans, and non-human primates. In the first part of this dissertation, we attempted reprogramming of three types of bovine somatic cells: fetal fibroblasts (bFFs), adult fibroblasts (bAFs), and bone marrow-derived mesenchymal stem cells (bMSCs), using six different culture conditions adapted from recent work in mice and humans. Using basic mouse reprogramming conditions, we did not succeed in inducing formation of ESC-like colonies in bovine somatic cells. The combination of 2i/LIF plus ALK5 inhibitor II and ascorbic acid, induced formation of colony-like structures with flat morphology, that occasionally produced trophoblast-like structures. These trophoblast-like vesicles did not appear when an inhibitor of Rho-associated, coiled-coil containing protein kinase 1 (ROCK) was included in the medium. We screened for expression of exogenous OSKM vector with RT-PCR and found upregulation of OSKM vector 24h after Dox was added to the medium; however, expression was sharply decreased on day 2 after Dox induction, and was not detectable after day 3. In a separate experiment, we induced reprogramming of bFF and bAFs using medium supplemented with 50% of medium conditioned by co-culture with the bovine trophoblast CT1 line. These cells expressed both OCT4 and the OSKM vector 24h after Dox induction. However, similar to our previous observations, both markers decreased expression until no signal was detected after day 3. In summary, we were unable to produce fully reprogrammed bovine iPSCs using mouse and human protocols, and the exact cause of our lack of success is unclear. It is possible that a different method of transgene expression could play a role in reprogramming. However, these ideas would be driven by a rather empirical reasoning, extrapolating findings from other species, and not contributing in our understanding of the particular differences of pluripotecy in ungulates. Our inability to produce bovine iPSCs, combined with the only partial reprogramming observed by others, justifies the need for in depth study of bovine pluripotency mechanisms, before meaningful attempts to reprogram bovine somatic cells to plutipotency are made. Therefore, we focused on getting a better understanding of bovine nuclear reprogramming. This would allow us to rationally target the specific requirements of potential bovine pluripotent cells. Cell fusion is a process that involves fusion of the membrane of two or more cells to form a multinucleated cell. Fusion of a somatic cell to an ESC is known to induce expression of pluripotency markers in the somatic nucleus. In the second part of this dissertation, we hypothesized that fusion of bFFs to mouse ESCs (mESCs) would induce expression of pluripotency markers in the bFF nucleus. We first optimized a cell fusion protocol based on the use of polyethylene glycol (PEG), and obtained up to 11.02% of multinucleated cells in bFFs. Next, we established a method to specifically select for multinucleated cells originated from the fusion of mESCs with bFFs (heterokaryons), using indirect immunofluorescence. With this in place, flow cytometry was used to select 200 heterokaryons which were further analyzed using RNA-seq. We found changes in bovine gene expression patterns between bFFs and heterokaryons obtained 24h after fusion. Focusing on the bovine transcriptome, heterokaryons presented upregulation of early pluripotency markers OCT4 and KLF4, as well as hypoxia response genes, contrasted with downregulation of cell cycle inhibitors such as SST. The cytokine IL6, known to increase survival of early embryos in vitro, was upregulated in heterokaryons, although its role and mechanism of action is still unclear. This indicates that the heterokaryon cell fusion model recapitulates several of the events of early reprogramming, and can therefore be used for further study of pluripotency in the bovine. The cell fusion model presented here can be used as a tool to characterize early changes in bovine somatic nuclear reprogramming, and to study the effect of different reprogramming conditions on the bovine transcriptome. / Ph. D.

This thesis is concerned with the topic of teaching histology at secondary schools. Histology is focused on the study of microscopic structure of tissue. By this it comprises the basic foundation stone for the studies of organs and organ systems not merely in humans, but also in other animals. The method of preparing the permanent histological slides, which has also been tested in practice, is thoroughly described in this thesis. The main goal of the practical part was to prepare a representative collection of the permanent histological sections through the organs of the house mouse (Mus muculus), to acquire its photo documentation as a base for an interactive histological atlas. House mouse (Mus musculus) was chosen as the model animal, because of its frequent utilization in various laboratory experiments and because this model mammal has the structure of tissues similar to human. The atlas consists of the photo documentation of histological sections through the following organs: lungs, skin, heart, thymus gland, spleen, epididymis, testicle, penis, spermatic sacs, striated muscle, heart muscle, smooth muscle, liver, tongue, pancreas, small intestine, large intestine, gall bladder, kidney, urinary bladder, urethra, cerebellum and hippocampus. The sections are stained with application of...

Les relations hôte-parasite sont complexes et soumettent les deux protagonistes à un ensemble de compromis aussi bien plastiques qu’évolutifs. D'un côté, bien que l’immunité soit indispensable pour la lutte contre les parasites, elle peut aussi causer de nombreux dégâts à l’hôte lors de la réponse et mener à des maladies inflammatoires. De l’autre côté, les parasites, bien que dotés de mécanismes d'évasion, vont être affectés par l'environnement inflammatoire de l'hôte. Cela pose la question des coûts et bénéfices que l'interaction entre ces deux protagonistes va avoir sur chacun d'eux lors de l'apparition de troubles inflammatoires chez l'hôte. Grâce à différentes approches expérimentales et bibliographiques nous avons pu montrer que l’immunopathologie est un trait qui perdure vraisemblablement grâce aux bénéfices de la réponse immunitaire dans la lutte contre les parasites. Par ailleurs, j’ai pu mettre en évidence qu’une inflammation altérait positivement les traits d’histoire de vie du parasite Heligmosomoides polygyrus aussi bien de façon plastique que lors de processus de sélection. Cependant, le parasite investit davantage dans l’immunomodulation et le camouflage lorsqu'il est confronté à cet environnement, laissant planer la question du coût de cette inflammation à long terme et sur plusieurs générations. / Host-parasite interactions are characterized by trade-offs that involve both plastic and microevolutionary responses. On one hand, while immunity is essential to fight parasites, it can also cause damage to the host, leading to autoimmunity and inflammatory diseases. On the other hand, parasites have to cope with the immune environnement provided by the host. This raises the question of the costs and benefits of the inflammatory response for the two partners of the interaction. With different experimental and literature-based approaches, I showed that immunopathology is a trait that likely persists because of the immediate benefits of the immune response in terms of protection against parasites. Furthermore, I was able to show that inflammation positively altered the life history traits of the gastrointestinal nematode Heligmosomoides polygyrus both plastically or after experimental evolution. However, the parasite invested more in immunomodulation and camouflage when facing an inflammatory environment, leaving open the question of the costs associated with an inflammatory environment over the entire lifespan of the parasite and/or across generations.

La naissance est l’une des étapes les plus délicates à laquelle les nouveau-nés mammifères doivent faire face. Le nouveau-né doit opérer des changements physiologiques et comportementaux pour s’adapter à l’environnement aérien, et l’un des premiers défis est d’ingérer du colostrum et du lait. Il est surprenant que la nature des stimuli et les mécanismes impliqués dans le déclenchement de la tétée soient encore mal connus, alors que la survie du nouveau-né est conditionnée par le succès de la première tétée. Par conséquent, cette étude a pour but de comprendre comment un nouveau-né immature et inexpérimenté réussit à s’orienter vers une tétine, à la saisir et à la téter de façon efficace? Cette question générale est posée chez la souris en focalisant sur : i) la nature des substrats chimiques utilisés par les souriceaux pour atteindre les tétines maternelles ; ii) la variation de la puissance attractive de ces substrats au cours du développement ; et iii) l’implication des effets de l’expérience dans l’établissement des réponses adaptatives précoces. Premièrement, nos résultats mettent en lumière que les odeurs mammaires de femelles allaitantes induisent plus d’approches et de saisies de la tétine chez les souriceaux que celles émanant de femelles non allaitantes. Deuxièmement, les odeurs de liquide amniotique et de lait déclenchent la première saisie orale de la tétine chez des souriceaux à la naissance, alors que les odeurs de salives maternelle et infantile n’induisent ce comportement qu’après une brève expérience de tétée. Troisièmement, les souriceaux âgés de 0, 2 et 6 jours postnatals (P), ayant eu une expérience de tétée, affichent une attraction sélective envers des odeurs de laits collectés en début de lactation plutôt qu’en fin de lactation, alors que les souriceaux plus âgés P15 ne montrent aucune réponse sélective envers ces odeurs. En résumé, certains substrats biologiques présents sur les tétines de femelles allaitantes sont immédiatement attractifs après la naissance, tandis que d’autres ont besoin d’être appris pour être réactogènes. Par conséquent, la réponse initiale de recherche de la tétine chez le souriceau est contrôlée par des processus d’apprentissage postnatal. A ce stade, l’implication de l’apprentissage prénatal et de processus prédisposés n’a pu être prouvée, bien qu’elle ne soit pas exclue. Ces résultats montrent des capacités d’apprentissage sophistiquées chez le souriceau nouveau-né / Birth is one of the most delicate periods mammalian infants have to deal with. Newborns have then to adapt physiologically and behaviorally to the aerial environment, and one of their first challenges is to ingest colostrum and milk. It is paradoxical that the survival of pups is conditioned by the success of this first suckling, and that we have so little understanding of the stimuli that underlie and promote it. Thus, the present work aims to contribute to answer how immature and naïve newborns do manage to orient to a nipple, to grasp it, and to suckle efficiently? This general issue will be addressed in the mouse in focusing on: i) the nature of the chemical substrates that newborn mice use to reach nipples; ii) whether the attractive potency of these substrates changes as a function of development; and iii) whether exposure effects underly the establishment of early adaptive responses? The results highlight that mammary odors of lactating females are more behaviorally active for newly born pups than those of non-lactating females. Secondly, amniotic and milk odors provoke the first nipple grasping in newborns right at birth, while maternal and pup salivary odors induce this behavior after short sucking experience. Thirdly, younger pups on postnatal day (P) 0, 2 and 6 (with sucking experience), display a selective orientation toward the odor of milk collected during early-lactation rather than to the odor of late-lactation milk, whereas older pups P15 do not exhibit any selective attraction to these odors. To summarize, some of the biological substrates which are present on a nursing mouse nipples are attractive immediately after birth, while some others need to be postnatally learned to be active. Thus, the initial nipple search response of mouse pups is controlled by processes involving postnatal learning. At this stage, the involvement of prenatal learning and of predisposed processes that do not depend on previous exposure effect are not conclusive, although not excluded. These results show highly sophisticated learning abilities in a newborn mammal

The term "personality" nowadays occurs more often not only in psychological studies of humans but also in animal studies. Studying of personality help us to define the behavioural characteristics which can vary within the age, sexes, species or enviroments. Behavioral experiments are used to detect these behavioral patterns and they can divide the animals into the different groups. The subject of our research became three populations of house mouse (Mus musculus sensu lato) which we tested in a series of experiments involving free exploration, forced exploration, hole- board test, test of vertical activity and Elevated plus-maze. These experiments should reveal wheter the mice differ in their behaviour through the context of sex, comensalism or subspecies. We found (with in excepcion of one test) that intrapopulation variability differences are very small but interpopulation differences purely increase in the cas of comensalism and effects of subspecies. Keywords: Mus musculus, comensalism, open fieldtest, Elevated plus-maze, Principal Component Analysis (PCA)