A methodology for ageing foetal and neonatal cattle is developed, involving radiographic examination of infant mandibles for early developmental stages in molariform teeth; tooth-wear methodologies are imprecise at this stage before wear commences. Known-age modern bovine foetal and neonate material are collected as a control assemblage for method development (n=73); six Neolithic to Norse era assemblages from Orkney are examined using the modified technique together with standard tooth-wear analysis and other methodologies. Foetal and died-at-birth material is diagnosed at most sites using the new technique, together with a range of other peri-natal age-groups. Ageing at this early stage is highly relevant in the diagnosis of milking as a palaeoeconomy: the accepted view is that unwanted (male) calves were slaughtered to maximise milk for human consumption, hence a surfeit of neonate calf remains, as at the study sites. The diagnosis of foetal and died-at-birth material challenges this view, suggesting that attritional causes may have contributed to deaths at this stage. Although milking was probably carried out at most of the study sites, this may have been combined with slaughter of cattle for meat in a pragmatic exploitation strategy. Literary research shows possible attritional causes of abortion and early death in calves, in particular dietary insufficiency in pregnant cows, microbial infections, and also inadequate colostrum uptake. Additionally, research is used to consider the challenges to health that early milking might have posed, to the calf as mentioned, but also to the cow, where three main health issues are highlighted: infertility, mastitis and lameness.

L'observation des nombreuses espèces au sein des métazoaires permet de rendre compte de leur formidable diversité de morphologies. Ces organismes complexes acquièrent leur plan d'organisation et leurs caractéristiques propres pendant le développement embryonnaire. Au cours de celui-ci, la morphogénèse des différentes structures anatomiques est contrôlée par des réseaux complexes de gènes intervenant dans des territoires et des moments précis. Comprendre quelles modifications des voies développementales au cours de l'évolution sont responsables de cette diversité constitue un champ important de la biologie moderne : l'évo-dévo. Pour comprendre ces modifications, il est important de pouvoir étudier ces changements sur des modèles facilement accessibles et qui possèdent une grande variabilité de formes. Les phanères des vertébrés répondent à ces critères : leur nombre, leur forme et leur taille ont été des caractères très variables au cours de la diversification des mammifères et les données déjà obtenues sur la souris permettent une expérimentation aisée. De nombreuses voies de signalisation sont impliquées dans le développement des phanères, mais une en particulier, impliquée spécifiquement dans le développement des appendices ectodermiques : la voie EDA, composée d'un ligand EDA, de son récepteur EDAR et d'un adaptateur spécifique EDARADD. Dans le but de mieux comprendre le rôle de la voie EDA au cours de l’évolution des mammifères, nous avons orienté ma thèse en 2 axes : le premier vise à étudier le rôle d’Edar dans le patterning de la dent chez la souris, et le second l’impact de la perte d’une des deux isoformes d’EDARADD au cours de l’évolution des mammifères. / The observation of the numerous metazoan species highlights their wonderful morphological diversity. These complex organisms got their body plan and their specific traits during embryonic development. During development, complex gene networks that are tightly regulated through space and time, control morphogenesis. Understanding which modifications of developmental pathways are responsible for the establishment of this diversity is one of the key questions of the biological field: Evo-Devo. To understand these modifications, it is crucial to study accessible models that are representative of this diversity. To do that, ectodermal appendages are a very good model: their number, size, and shape are highly variable during mammals diversification and data had already shown that they exhibit natural variation. Numerous signaling pathways are implicated in their development but one is very specific to them: the Eda pathway and present an big interest for the study of their evolution. To better understand the role of the Eda pathway during mammals evolution, I orientated my thesis in two part: the first one study the impact of Edar in the establishment of the mouse dental tooth row and the second the impact of gain/loss of protein isoforms of the adaptor Edaradd on the evolution of this pathway.

(v anglickém jazyce) Development of tooth primordia in mice is an important model for study of odontogenesis. Several dental rudiments develop during the mouse embryogenesis. These structures develop in functional teeth in their phylogenetically older relatives. Similarly, we can initiate growth of teeth from these germs in some mutant mice. In my diploma thesis we have focused on the importance of rudimentary structures with odontogenic potential in postnatal individuals. As a model of development, we have chosen a cell population originating from rudimentary primordia MS (mesial segment) that develops in diastema of the lower jaw during the embryonic day 12.5. Using the inducible Cre-lox technology we have marked the cells which are part of the signal domain of primordia at this time. As a marker of these cells we have used gene Shh. We have found out that these cells persist prenataly and also postnatally. Further we have isolated this cell area and we have tested it using a variety of methods. We have shown that in the cells of postnatal individual are expressed markers of stem cells (Sox2, Bmi1, Gli1) and also genes for major enamel matrix structural proteins: ameloblastin and amelogenin. The same stem cell markers are also expressed in vitro culture of the isolated cells. This cell population...

Le développement dentaire est essentiellement étudié chez la souris, modèle mammifère le plus commun en biologie. Cependant, contrairement à la majorité des mammifères, les souris ne remplacent pas leurs dents. Ainsi, les mécanismes impliqués dans le remplacement dentaire mammalien sont encore inconnus. Au cours de cette thèse, nous nous sommes intéressés au développement et remplacement dentaire mammalien en utilisant le lapin Oryctolagus cuniculus comme modèle d’étude. Le lapin étant déjà séquencé, utilisé en recherche biomédicale avec une période de gestation courte et remplaçant ses dents, il semblait être un modèle pertinent en odontologie. Le lapin était un modèle méconnu du point de vue du développement dentaire, nous avons donc d’abord réalisé une étude histo-morphologique afin de caractériser la mise en place des dents déciduales et permanentes. Des reconstructions 3D des tissus mous ont été réalisés à différents stades embryonnaires afin d’obtenir une chronologie du développement et remplacement dentaire. Cette chronologie commence aux premières observations morphologiques de l’initiation du développement des premières dents jusqu’à la minéralisation des dernières dents à se développer.Puis, suite à l’identification dans la bibliographie de gènes candidats potentiellement impliqués dans le remplacement dentaire, nous avons étudiés les profils d’expressions de ces gènes afin de mieux comprendre la régulation spatio-temporelle du remplacement dentaire chez le lapin. Nous avons ensuite replacé nos résultats sur le développement chez le lapin dans un contexte évolutif. Ainsi, nous avons réalisé une étude d’anatomie comparée chez les lagomorphes actuels et quelques fossiles afin d’identifier des variations morphologiques dentaires au cours de leur histoire évolutive. Nous nous sommes particulièrement intéressé à la mise en place des cuspides au cours de l’odontogénèse ainsi qu’aux variations de la surface occlusale des dents supérieure tout au long de la vie des lapins et autres lagomorphes. En comparant les variations au cours de l’évolution avec celles observées lors de l’ontogénie dentaire chez le lapin nous avons identifié des processus d’hétérochronies du développement. Nous avons ainsi montré que la molaire actuelle du lapin suit un processus de péramorphose, donc de surdéveloppement, en comparaison aux lagomorphes fossiles. Le lapin est ainsi un modèle animal prometteur en biologie du développement et en évolution afin de mieux comprendre la mise en place du remplacement dentaire mammalien ainsi que les variations de forme dentaire au cours de l’évolution des mammifères. / Tooth development is essentially studied in mice, the favorite mammalian model in biology. However, mice do not replace their teeth on contrary to numerous mammals. So, the mechanisms involved in mammalian tooth replacement are still unknown. In this thesis, we focused on mammalian dental development and replacement using the European rabbit Oryctolagus cuniculus as animal model. The European rabbit is already sequenced, used in biomedical field, has a short gestation time and replaced its teeth, so rabbit seemed to be a relevant model in dental research.Rabbit dental development was not defined, so we first performed a histo-morphological study to characterize the development of deciduous and permanent teeth. 3D soft tissue reconstructions were performed at different embryonic stages to obtain a chronology of tooth development and replacement. This chronology begins with the first morphological observations of the initiation of the development of the first tooth until the mineralization of the last tooth to develop.Then, we identified in the bibliography candidate genes potentially involved in dental replacement. We studied the expression profiles of these genes in order to better understand the spatio-temporal regulation of tooth replacement in rabbits.We then returned our results to rabbit development in an evolutionary context. Thus, we performed a comparative anatomy study in the current lagomorphs and some fossils in order to identify dental morphological variations during their evolutionary history. We were particularly interested in the setting of cusps during odontogenesis as well as in the variations of the occlusal surface of the upper cheek teeth throughout the life of rabbits and other lagomorphs. Comparing changes during evolution with those observed during dental ontogeny in rabbits allow us to identify heterochronous processes of development. We have shown that the current molar rabbit follows a process of peramorphosis, so an overdevelopment compared to fossil lagomorphs.The rabbit is thus a promising animal model in developmental and evolutionary biology to better understand the implementation of mammalian tooth replacement and tooth shape variations during mammalian evolution.

A methodology for ageing foetal and neonatal cattle is developed, involving radiographic examination of infant mandibles for early developmental stages in molariform teeth; tooth-wear methodologies are imprecise at this stage before wear commences. Known-age modern bovine foetal and neonate material are collected as a control assemblage for method development (n=73); six Neolithic to Norse era assemblages from Orkney are examined using the modified technique together with standard tooth-wear analysis and other methodologies. Foetal and died-at-birth material is diagnosed at most sites using the new technique, together with a range of other peri-natal age-groups. Ageing at this early stage is highly relevant in the diagnosis of milking as a palaeoeconomy: the accepted view is that unwanted (male) calves were slaughtered to maximise milk for human consumption, hence a surfeit of neonate calf remains, as at the study sites. The diagnosis of foetal and died-at-birth material challenges this view, suggesting that attritional causes may have contributed to deaths at this stage. Although milking was probably carried out at most of the study sites, this may have been combined with slaughter of cattle for meat in a pragmatic exploitation strategy. Literary research shows possible attritional causes of abortion and early death in calves, in particular dietary insufficiency in pregnant cows, microbial infections, and also inadequate colostrum uptake. Additionally, research is used to consider the challenges to health that early milking might have posed, to the calf as mentioned, but also to the cow, where three main health issues are highlighted: infertility, mastitis and lameness. / The attached files include the Landscape pages and appendices V and VI. Not included are the jpeg Mandible files. A cover sheet was not available.

CHAPER I: Through a review of the literature, this article discusses the genetic mechanisms that control tooth morphogenesis. Emphasis is placed upon the structure and function of some key molecules that participate in interactions between its epithelial-mesenchimal components. In this paper we will can understand the mechanisms that control tooth morphogenesis and the dentistry should pay special attention to possible consequences of tooth number anomalies. CHAPER II: The analysis of DNA is widely employed in the genetic studies. Human DNA in most cases is performed with samples obtained from peripheral blood. The use of buccal epithelial cells as a source of DNA for PCR amplifications has several advantages over blood sampling. In the present study our objective was to standardize DNA extraction from an oral swab, using a simple method. To test DNA quality, we amplified the exons 2 of MSX1 gene and the promoter region of LEF1 gene to patients with hypodontia. In conclusion, we standardized a simple DNA. extraction of oral cells, which presented lower costs and faster results, indicating to that DNA from oral brushes/swabs are a reliable source for genetic studies. The quantity and quality of extracted DNA was shown to be adequate for PCR and polymorphism analyses. CHAPER III: Hypodontia, the congenital absence of one or a few teeth, is one of the most common alterations of the human dentition. The most common permanent missing teeth are the third molars, second premolars, and maxillary lateral incisors. Although hypodontia does not represent a serious public health problem, it may cause masticatory and speech dysfunctions and esthetic problems. In human the participation of MSX1 gene in craniofacial development have been evidenced by the studes that showed mutations in this gene. Hypodontia were shown to be caused by mutations in the MSX1 gene in human however, the mutation in the MSX1 gene cannot explain all types of tooth agenesis. Our data suggest that polymorphisms in MSX1 gene are associated with hypodontia. / CAPITULO I: Este artigo apresenta uma revisão bibliográfica sobre as evidências mais atuais sobre os aspectos genéticos da formação dos dentes. São abordadas nesse artigo as principais moléculas envolvida na interação epitélio-mesênquima, responsável pela formação da estrutura dental. O objetivo é contribuir para um melhor entendimento da expressão genética envolvida na formação do dente, bem como auxiliar na prática odontológica, procurando despertar a atenção do profissional para o conhecimento científico e facilitar assim a identificação de possíveis problemas relacionados à formação dos elementos dentais. CAPITULO II: A análise do DNA é largamente usada em estudos genéticos. O DNA humano, em muitos casos, é obtido através de amostras de sangue periférico. O uso de células descamadas da mucosa oral, como fonte de DNA para amplificação por PCR, tem apresentado muitas vantagens. Nesse estudo, nosso objetivo foi padronizar extração de DNA de células obtidas da mucosa oral, usando um novo método. Para testar a qualidade do DNA, nós amplificamos o segundo éxon do gene MSX1 em pacientes com hipodontia. Criamos um novo método de extração de DNA através de células da mucosa oral, que apresenta baixo custo e rápidos resultados, indicando que o DNA dessas células,quando extraídos por essa técnica, é suficiente para estudos genéticos. O DNA extraído mostrou-se adequado em quantidade e qualidade, para estudos de PCR e análises de polimorfismos. CAPITULO III: Hipodontia, a ausência congênita de dentes, é uma das alterações mais comuns na dentição humana. Vários dentes podem estar ausentes porém, os mais comuns são os terceiros molares, segundo pré-molares e incisivos laterais superiores. Embora essa alteração de número não represente um problema de saúde pública, ela pode causar disfunções mastigatórias e problemas estéticos graves. Nos humanos, o papel do gene MSX1 no desenvolvimento crânio facial tem sido esclarecido em estudos que identificaram mutações nesses genes, associadas a alterações da normalidade. Mutações-polimorfismos no gene MSX1 têm sido relatadas como responsáveis pela agenesia dental, no entanto, mutações neste gene não explicam todas as formas dessa alteração. Nossos resultados sugerem que polimorfismos no gene MSX1 estão associados com a hipodontia. / Doutor em Genética e Bioquímica