Ο δάκος της ελιάς, Bactrocera oleae, αποτελεί το κυριότερο παράσιτο του καρπού της ελιάς. Λόγω της μεγάλης οικονομικής σημασίας της ελιάς, ιδιαίτερα για τις Μεσογειακές χώρες, ο αποτελεσματικότερος έλεγχος του εντόμου αυτού είναι απαραίτητος. Η εφαρμογή μεθόδων ολοκληρωμένης διαχείρισης του παρασίτου αυτού, μεθόδων δηλαδή φιλικών προς το περιβάλλον, επιβάλλει την καλύτερη γνώση της βιολογίας του και ιδιαίτερα της γενετικής και της γενετικής δομής των φυσικών πληθυσμών του. Στα πλαίσια αυτά, στόχος της παρούσας μελέτης ήταν η ανάπτυξη στο δάκο DNA δεικτών, των μικροδορυφόρων, οι οποίοι είναι άφθονοι στα γονιδιώματα και επιπλέον υψηλά πολυμορφικοί, για την ανάλυση φυσικών πληθυσμών του. Για την απομόνωση των μικροδορυφορικών δεικτών ακολουθήθηκαν τρεις διαφορετικές στρατηγικές: η κατασκευή και διαλογή γονιδιωματικών βιβλιοθηκών, η κατασκευή εμπλουτισμένων σε μικροδορυφόρους βιβλιοθηκών και ο έλεγχος ζευγών εκκινητών που είχαν σχεδιαστεί για την ενίσχυση μικροδορυφόρων στα συγγενικά είδη B. tyroni και C. capitata για το κατά πόσο ενισχύουν τις αντίστοιχες περιοχές και στο γονιδίωμα του εντόμου B. oleae (cross-species amplification). Από τις βιβλιοθήκες απομονώθηκαν συνολικά 69 κλώνοι που περιείχαν μικροδορυφόρους. Ακολούθησε ο σχεδιασμός ζευγών εκκινητών στις μοναδικές περιοχές που περιβάλλουν τους μικροδορυφόρους. Συνολικά σχεδιάστηκαν 42 ζεύγη εκκινητών. Οι εκκινητες αυτοί ελέγχθηκαν για το αν ενισχύουν το αναμενόμενο προϊόν. Ο έλεγχος έγινε με PCR και ηλεκτροφόρηση των προϊόντων σε πήκτωμα αγαρόζης. Παράλληλα ελέγχθηκαν είκοσι ζεύγη εκκινητών που είχαν σχεδιαστεί για μικροδορυφόρους του εντόμου B. tryoni και 42 ζεύγη εκκινητών που είχαν σχεδιαστεί για μικροδορυφόρους του εντόμου C. capitata. Οι τρεις διαδικασίες απομόνωσης έδωσαν συνολικά 67 λειτουργικά ζεύγη εκκινητών. Στη συνέχεια ελέγχθηκε μέσω PCR ο πολυμορφισμός των εκκινητών αυτών και το αν ενισχύουν διακριτά αλληλόμορφα, με μήτρα DNA γενετικό υλικό εννέα ατόμων. Τα προϊόντα της PCR αναλύθηκαν σε πήκτωμα ακρυλαμιδίου. Από τα 49 ζεύγη εκκινητών που ελέχθησαν, τα 28 έδωσαν πολύ καθαρό σήμα, ενώ τα 25 από αυτά βρέθηκαν πολυμορφικά. Μια πρώτη πληθυσμιακή ανάλυση έγινε με τη χρησιμοποίηση 24 από αυτούς τους δείκτες για την ανάλυση μικρού δείγματος ατόμων. Βασικός σκοπός της ανάλυσης αυτής ήταν να ελεγχθεί η ποιότητα των δεικτών και οδήγησε στον αποκλεισμό έξι δεικτών: ο ένας ήταν μονομορφικός, ένας δεύτερος έδειξε ασθενή ενίσχυση και μικρή επαναληψιμότητα και άλλοι τέσσερις εμφάνισαν απόκλιση από την ισορροπία κατά H-W, πιθανότητα λόγω της παρουσίας null αλληλομόρφων. Δώδεκα από τους υπόλοιπους δεκαοκτώ δείκτες χρησιμοποιήθηκαν για μια εκτεταμένη ανάλυση των πληθυσμών του δάκου στην Ευρωπαϊκή πλευρά της λεκάνης της Μεσογείου. Αναλύθηκαν δεκαεννέα δείγματα, μεγέθους εννέα ως πενήντα ατόμων, από έξι διαφορετικές χώρες (Ελλάδα, Κύπρος, Τουρκία, Ιταλία, Ισπανία και Πορτογαλία). Η ανάλυση αποκάλυψε σχετικά μικρές γενετικές αποστάσεις, που έδειχναν όμως μια στατιστικά σημαντική διαφοροποίηση σε τρεις υποπληθυσμούς. Ο πρώτος αποτελείτο από τα δείγματα της Κύπρου, ο δεύτερος από τα δείγματα Ελλάδας, Τουρκίας και Ισπανίας και ο τρίτος από τα δείγματα της Ιβηρικής χερσονήσου. Οι στατιστικές αναλύσεις που έγιναν έδειξαν τη σημαντική επίδραση της γεωγραφικής απόστασης στη δημιουργία αυτών των ομαδοποιήσεων. Οι τρεις αυτές ομάδες χαρακτηρίζονται από διαφορά και στο επίπεδο του πολυμορφισμού, εμφανίζοντας μια καθαρή μείωσή του από την Ανατολή προς τη Δύση. Η μείωση αυτή είναι στατιστικά σημαντική και με βάση την υπόθεση ότι η πορεία εποικισμού ενός είδους συνοδεύεται από μείωση του πολυμορφισμού, δίνει σημαντικές ενδείξεις για μία προς Δυσμάς πορεία εποίκισης του είδους στον Ευρωπαϊκό χώρο, με πρώτο κέντρο εξάπλωσης την Ανατολική λεκάνη της Μεσογείου. Τα πειράματα δια-ειδικής ενίσχυσης έδειξαν στενές φυλογενετικές σχέσεις των ειδών που εξετάστηκαν και κυρίως μεταξύ B. tryoni - B. oleae και B.oleae - C. capitata. Τα πειράματα αυτά υποστηρίζουν την χρησιμότητα των δεικτών που απομονώθηκαν σε ενδεχόμενες φυλογενετικές μελέτες στα είδη αυτά, καθώς και σε άλλα συγγενικά είδη. Η εύρεση ενός μεγάλου ποσοστού συντηρημένων μικροδορυφόρων σε είδη που έχουν διαχωριστεί εδώ και πολλά εκατομμύρια χρόνια ενισχύει την υπόθεση ότι οι μικροδορυφόροι δεν είναι γενετικό υλικό χωρίς ρόλο (junk DNA), αλλά επιτελούν συγκεκριμένες λειτουργίες στο γονιδίωμα. Η ανάλυση πολυμορφισμού που έγινε στις ατομικές διασταυρώσεις ατόμων από εργαστηριακούς ήταν ιδιαίτερα ενθαρρυντική. Ο υψηλός πολυμορφισμός που βρέθηκε δείχνει την δυνατότητα χρησιμοποίησης τους και στη γενετική χαρτογράφηση του είδους. / The olive fruit fly, Bactrocera oleae, is the main pest of the olive fruit. Because of its great economic importance, especially for the Mediterranean countries, there is a need for a more effective control method. The application of an integrated, environmental friendly, management of this pest requires a better knowledge of its biology and of the genetic structure of its natural populations. The aim of the present study was the development of DNA microsatellite markers for the analysis of the natural populations of the olive fruit fly. These markers are abundant in the genome of any species studied so far and highly polymorphic. Three different strategies were used for the isolation of microsatellite markers. The first was the construction and screening of genomic libraries of the insect, the second was the construction of genomic libraries, enriched for microsatellites and the third was the use of primer pairs that were designed for the amplification of microsatellite markers in the closely related species Bactrocera oleae and Ceratitis capitata(cross-species amplification). A total of 69 microsatellite containing clones were isolated from libraries. The next step was the design of primer pairs in the microsatellite flanking sequences. A total of 42 primer pairs was designed and tested for their abillty to amplify the expected product. Test was performed through PCR and analysis of the PCR products through electrophoresis on agarose gel. Twenty primer pairs designed for the amplification of Bactrocera tryoni’ s microsatellites and 42 primer pairs designed for the amplification of Ceratitis capitata microsatellites were also tested. All three strategies gave 67 primer pairs that amplified the expected product. The degree of polymorphism of these primer pairs and their ability to amplify easily resolvable alleles was tested through PCR with DNA template of nine individuals. PCR products were analysed through polyacrylamide gel electrophoresis. Twenty eight out of the 49 primer pairs tested produced clear bands and twenty five of them were polymorphic. A small scale population analysis was then performed, using tventy four of the markers available. The main purpose of this analysis was to demonstate the quality of the markers and lead to the exclusion of six markers: one of them was monomorphic, another didn’t show reproducible results and four more showed deviations from H-W equilibrium, probably because of the presence of null alleles. Twelve of the remaining loci were used in a large scale analysis of B. oleae’s populations in the European part of the Mediterranean basin. Nineteen samples, varying from nine to fifty individuals, were analysed. These samples were collected from six different countries (Greece, Cyprus, Turkey, Italy, Spain and Portugal). The analysis revealed relatively low genetic distances, which, however, demonsrated a statistically important differentiation of the samples in three subpopulations. The first consisted of the samples from Cyprus, the second of the samples from Greece, Turkey and Italy and the third of the samples from the Iberian Peninsula. The statistical analyses performed showed the statistically important contribution of geographic distance to the generation of genetic distance. These three groups of samples also demonstrate a clear loss of polymorphism towards West. This loss is statistically important and, if we take into account the hypothesis that the colonization process of a species is followed by a loss in polymorphism, it suggests a colonization process of the olive fruit fly towards West in the European part of the Mediterranean basin, with a first expansion area in the Eastern part of the Mediterranean basin. Cross-species amplification experiments indicate close phylogenetic relationships among the species studied, mainly between B. tryoni-B. oleae and B. oleae-C. capitata. These results support the usefulness of the markers isolated in phylogenetic studies in these species, as well as in other, closely related species. The identification of a high percentage if conserved microsatellites in species that have been well separated for millions of years is in agreement with the hypothesis that microsatellites are not useless genomic regions (junj DNA) but they perform specific functions in the genome. Polymorphism analysis in the crosses of individuals from laboratory strains was very encouraging. The high degree of polymorpism showes that they can be used in genetic mapping of the species

Submitted by Luciana Sebin (lusebin@ufscar.br) on 2016-10-07T18:32:03Z No. of bitstreams: 1 DissPPP.pdf: 1959628 bytes, checksum: 65b4132f7bd4b59eb075f69f2b29aac8 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-10-13T19:51:11Z (GMT) No. of bitstreams: 1 DissPPP.pdf: 1959628 bytes, checksum: 65b4132f7bd4b59eb075f69f2b29aac8 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-10-13T19:51:21Z (GMT) No. of bitstreams: 1 DissPPP.pdf: 1959628 bytes, checksum: 65b4132f7bd4b59eb075f69f2b29aac8 (MD5) / Made available in DSpace on 2016-10-13T19:51:31Z (GMT). No. of bitstreams: 1 DissPPP.pdf: 1959628 bytes, checksum: 65b4132f7bd4b59eb075f69f2b29aac8 (MD5) Previous issue date: 2015-08-28 / The black lion tamarin (Leontopithecus chrysopygus) is one of the most endangered neotropical primates and the historical causes that led them to the brink of extinction are closely related to the history of the Atlantic Rainforest. Among its main threats are the fragmentation of their habitat, the small number of his surviving populations and the isolation of them, which directly affect the genetic structure of these populations. The use of genetic analysis and molecular markers for the wildlife conservation have been growing over the past few years with the advent of genetic and molecular technologies and bioinformatics, allowing the establishment of rapid diagnosis of diseases and many genetic and ecological parameters such as migration rate, population size, genetic diversity, kinship relations. Among the main molecular markers are microsatellite that consist of short DNA sequences tandemly repeated composed of 1 to 6 bases pairs widely distributed in eukaryotic and prokaryotic genomes. Characteristics like codominance and high level of polymorphism make microsatellites an important tool to measure the loss of genetic diversity and recent changes in genetic structure of populations, and for use in forensic investigations. Among the methods used for the isolation of such markers, is the in silico mining for species with available genetic data. In the present study, we identified 60 tetranucleotide microsatellite loci have been identified in the genome of common marmoset (Callithrix jacchus) through data mining conducted in the genome of this species. Primer pairs were designed and tested in black lion tamarin, since this kind does not have any genomic data available. Of the 60 loci tested, 87% had successful amplification of DNA samples from 10 captive animals. PCR products were analyzed on agarose gel and 13 loci were genotyped in automatic sequencer ABI3730XL. The Geneious version 8.1.6 software was used for genotyping. Only four loci showed polymorphism, observed two alleles per locus. This low polymorphism may be associated with the origin of the captive colonies. / O mico-leão-preto (Leontopithecus chrysopygus) é um dos mais ameaçados primatas neotropicais e as causas históricas que o levaram à beira da extinção estão intimamente relacionadas à história da Mata Atlântica. Entre suas principais ameaças estão a fragmentação de seu habitat, o número reduzido de suas populações sobreviventes e o isolamento das mesmas, as quais afetam diretamente a estrutura genética dessas populações. O uso de análises genéticas e marcadores moleculares a favor da conservação da vida silvestre vêm crescendo ao longo dos últimos anos com o surgimento das tecnologias genéticas, moleculares e da bioinformática, possibilitando o estabelecimento do rápido diagnóstico de doenças e de muitos parâmetros genéticos e ecológicos, como taxa de migração, tamanho populacional, diversidade genética, relações de parentesco. Dentre os principais marcadoes estão os microssatélites que consistem em pequenas sequências de DNA repetidas em tandem compostas de 1 a 6 pares de base amplamente distribuídas nos genomas eucarióticos e procarióticos. Suas características de codominância e alto nível de polimorfismos fazem desses marcadores ferramentas importantes para estimativas de perda de variabilidade e de mudanças recentes na estruturação genética de populações, além do uso em investigações forenses. Dentre os métodos utilizados para o isolamento de tais marcadores, está a mineração in silico para espécies com dados genéticos disponíveis. No presente estudo, identificamos 60 microssatélites tetranucleotídicos no genoma do saguí-de-tufo-branco (Callithrix jacchus) através de Data Mining realizados no genoma desta espécie. Pares de primers foram desenhados e testados em Leontopithecus chrysopygus, uma vez que esta espécie não possui dados genômicos disponíveis. Dos 60 locos testados, 87% tiveram sucesso de amplificação em amostras de DNA provenientes de 10 animais de cativeiro. Os produtos de PCR foram analisados em gel de agarose e 13 locos foram genotipados em sequenciador automático ABI3730XL. O programa Geneious versão 8.1.6 foi utilizado para genotipagem. Somente quatro locos demonstraram polimorfismo, sendo observados dois alelos por loco. Esse baixo polimorfismo pode estar associado à origem das colônias de cativeiro.

Complex phenotypes are traits which are influenced by many factors, and not just a single gene, as for classical Mendelian traits. The brain, and its resultant behaviour, gives us a large subset of complex phenotypes to examine. Variation in these traits is affected by a range of different influences, both genetic and environmental, including social interactions and the effects of parents. Systems-genetics provides us with a framework in which to examine these complex traits, seeking to connect genetic variants to the phenotypes they cause, through intermediate phenotypes, such as gene expression and protein levels. This approach has been developed to exploit and analyse massive data sets generated for example in genomics and transcriptomics. In the first half of this thesis, I combine genetic linkage data from the BXD recombinant inbred mouse panel with genome-wide association data from humans to identify novel candidate genes, and use online gene annotations and functional descriptions to support these candidates. Firstly, I discovered MGST3 as a novel regulator of hippocampus size, which may be linked to neurodegenerative disorders. Secondly, I identified that CMYA5, MCTP1, TNR and RXRG are associated with mouse anxiety-like phenotypes and human bipolar disorder, and provide evidence that MCTP1, TNR and RXRG may be acting via inter-cellular signalling in the striatum. The second half of this thesis uses different cross-fostering designs between genetically variable BXD lines and the genetically uniform C57BL/6J strain to identify indirect genetic effects and the loci underlying them. With this, I have found novel loci expressed in mothers that alter offspring behaviour, novel loci expressed in offspring affecting the level of maternal care, and novel loci expressed in offspring, which alter the behaviour of their nestmates, as well as the level of maternal care they receive. Further I provide evidence of co-adaptation between maternal and offspring genotypes, and a positive indirect genetic effect of offspring on their nestmates, supportive of a role for kin selection. Finally, I demonstrate that the BXD lines can be used to investigate genes with parent-of-origin dependent expression, which have an indirect genetic effect on maternal care. In conclusion, this thesis identifies a number of novel loci, and in some cases genes, associated with complex traits. Not only are these techniques applicable to other phenotypes and other questions, but the candidates I identify can now be examined further in vitro or in vivo.

Neu auftretende Viruskrankheiten zoonotischen Ursprungs stellen eine zunehmende Gefahr für die globale Gesundheit dar. Wie das unerwartete Auftreten von dem Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Ende 2019 zeigte, gibt es allerdings trotz jahrelanger intensiver Forschung Verständnislücken, wo und wann mit Tieren assoziierte Krankheitserreger auftauchen, oder durch welche evolutionären Vorgänge diese Übertragungen möglich werden. Diese Arbeit befasst sich mit dem Thema der genetischen Adaptation von Viren bevor und nachdem ein Wirtswechsel stattgefunden hat und leistet damit einen Beitrag zum besseren Verständnis von Wirtswechselprozessen und ihren zugrundeliegenden Mechanismen. / Emerging viral diseases of zoonotic origin pose an increasing threat to global health. Despite intense research, we do not understand where and when animal-associated pathogens emerge, or by what evolutionary processes these transmissions become possible; best illustrated by the unexpected emergence of the Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the end of 2019. This thesis is concerned with the topic of viral genetic adaptation before and after cross-species transmissions, contributing to a better understanding of host switching processes and their underlying mechanisms.

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No description available.

Gene expression regulation is dynamic and specific to various factors such as developmental stages, environmental conditions, and stimulation of pathogens. Nowadays, a tremendous amount of transcriptome data sets are available from diverse species. This trend enables us to perform comparative transcriptome analysis that identifies conserved or diverged gene expression responses across species using transcriptome data. The goal of this dissertation is to develop and apply approaches of comparative transcriptomics to transfer knowledge from model species to non-model species with the hope that such an approach can contribute to the improvement of crop yield and human health. First, we presented a comprehensive method to identify cross-species modules between two plant species. We adapted the unsupervised network-based module finding method to identify conserved patterns of co-expression and functional conservation between Arabidopsis, a model species, and soybean, a crop species. Second, we compared drought-responsive genes across Arabidopsis, soybean, rice, corn, and Populus in order to explore the genomic characteristics that are conserved under drought stress across species. We identified hundreds of common gene families and conserved regulatory motifs between monocots and dicots. We also presented a BLS-based clustering method which takes into account evolutionary relationships among species to identify conserved co-expression genes. Last, we analyzed single-cell RNA-seq data from monocytes to attempt to understand regulatory mechanism of innate immune system under low-grade inflammation. We identified novel subpopulations of cells treated with lipopolysaccharide (LPS), that show distinct expression patterns from pro-inflammatory genes. The data revealed that a promising therapeutic reagent, sodium 4-phenylbutyrate, masked the effect of LPS. We inferred the existence of specific cellular transitions under different treatments and prioritized important motifs that modulate the transitions using feature selection by a random forest method. There has been a transition in genomics research from bulk RNA-seq to single-cell RNA-seq, and scRNA-seq has become a widely used approach for transcriptome analysis. With the experience we gained by analyzing scRNA-seq data, we plan to conduct comparative single-cell transcriptome analysis across multiple species. / Doctor of Philosophy / All cells in an organism have the same set of genes, but there are different cell types, tissues, organs with different functions as the organism ages or under different conditions. Gene expression regulation is one mechanism that modulates complex, dynamic, and specific changes in tissues or cell types for any living organisms. Understanding gene regulation is of fundamental importance in biology. With the rapid advancement of sequencing technologies, there is a tremendous amount of gene expression data (transcriptome) from individual species in public repositories. However, major studies have been reported from several model species and research on non-model species have relied on comparison results with a few model species. Comparative transcriptome analysis across species will help us to transform knowledge from model species to non-model species and such knowledge transfer can contribute to the improvement of crop yields and human health. The focus of my dissertation is to develop and apply approaches for comparative transcriptome analysis that can help us better understand what makes each species unique or special, and what kinds of common functions across species have been passed down from ancestors (evolutionarily conserved functions). Three research chapters are presented in this dissertation. First, we developed a method to identify groups of genes that are commonly co-expressed in two species. We chose seed development data from soybean with the hope to contribute to crop improvement. Second, we compared gene expression data across five plant species including soybean, rice, and corn to provide new perspectives about crop plants. We chose drought stress to identify conserved functions and regulatory factors across species since drought stress is one of the major stresses that negatively impact agricultural production. We also proposed a method that groups genes with evolutionary relationships from an unlimited number of species. Third, we analyzed single-cell RNA-seq data from mouse monocytes to understand the regulatory mechanism of the innate immune system under low-grade inflammation. We observed how innate immune cells respond to inflammation that could cause no symptoms but persist for a long period of time. Also, we reported an effect of a promising therapeutic reagent (sodium 4-phenylbutyrate) on chronic inflammatory diseases. The third project will be extended to comparative single-cell transcriptome analysis with multiple species.

Advances in genomics allow researchers to measure the complete set of transcripts in cells. These transcripts include messenger RNAs (which encode for proteins) and microRNAs, short RNAs that play an important regulatory role in cellular networks. While this data is a great resource for reconstructing the activity of networks in cells, it also presents several computational challenges. These challenges include the data collection stage which often results in incomplete and noisy measurement, developing methods to integrate several experiments within and across species, and designing methods that can use this data to map the interactions and networks that are activated in specific conditions. Novel and efficient algorithms are required to successfully address these challenges. In this thesis, we present probabilistic models to address the set of challenges associated with expression data. First, we present a novel probabilistic error correction method for RNA-Seq reads. RNA-Seq generates large and comprehensive datasets that have revolutionized our ability to accurately recover the set of transcripts in cells. However, sequencing reads inevitably contain errors, which affect all downstream analyses. To address these problems, we develop an efficient hidden Markov modelbased error correction method for RNA-Seq data . Second, for the analysis of expression data across species, we develop clustering and distance function learning methods for querying large expression databases. The methods use a Dirichlet Process Mixture Model with latent matchings and infer soft assignments between genes in two species to allow comparison and clustering across species. Third, we introduce new probabilistic models to integrate expression and interaction data in order to predict targets and networks regulated by microRNAs. Combined, the methods developed in this thesis provide a solution to the pipeline of expression analysis used by experimentalists when performing expression experiments.

Ce travail visait à évaluer le potentiel d’une méthodologie innovante récemment adaptée à la Souris sur la base de tests neuropsychologiques utilisés en clinique humaine. Après optimisation de 3 tâches (PAL, VMCL, PVD) ciblant différentes fonctions cognitives chez l’animal, nos études ont établi : (1) l’existence possible d’interférences proactives lors d’apprentissages consécutifs ; (2) l’absence de déficit d’acquisition chez les souris de la lignée Tg2576 (modèle transgénique de la maladie d’Alzheimer), quelle que soit l’étendue de la charge amyloïde ; (3) l’implication spécifique du striatum dorsal lors de l’acquisition des tâches de VMCL et PAL, et celle de l’hippocampe lors du rappel de cette dernière tâche. Ces derniers résultats suggèrent qu’en dépit des efforts déployés pour s’assurer du caractère translationnel d’une tâche cognitive dans le paradigme du touchscreen, certaines adaptations inhérentes à chaque espèce influencent profondément les bases neurobiologiques associées. / This thesis work aimed to specify the potential of an innovative methodology latterly adapted in mice from neuropsychological tasks used in Humans. After the optimization of 3 assays (PAL, VMCL, PVD) taxing various cognitive functions in animals, different behavioral studies have gradually revealed: (1) the putative existence of proactive interferences over consecutive learnings in touchscreen tasks; (2) no acquisition deficit in Tg2576 mice (a transgenic model of Alzheimer’s Disease) in these paradigms, whatever the amyloid load considered; (3) the specific involvement of the dorsal striatum during the acquisition of VMCL and PAL tasks and the key role of the hippocampus during the recall of the latter task. As exemplified by the PAL task, our results suggest that despite momentous efforts in order to ensure the translational feature of touchscreen cognitive tasks, certain adaptations inherent to each species deeply influence the nature of underlying neurobiological substrates.

Die Leber ist das am häufigsten von Metastasen betroffene Organ und kann daher als Modellorgan für metastatische Invasion dienen. Aus diesem Grund war es das Ziel dieser Dissertation Genexpressionsprofile zu verstehen und metastasierungs- sowie invasionsassoziierte Gene zu identifizieren. Differentielle Genexpression wurde in drei Systemen überprüft: Einem syngenen Mausmodell, einem Xenograftmodell sowie in fünf Gewebeproben von Patienten. Genexpressionprofile des syngenen Mausmodells und der Patientenproben zeigten, dass man die Invasionsfront als Ganzes betrachten, um möglichst viele über-lappende Gene zu finden. Globale Genexpressionstudien, die auf den Wirtsteil der Invasionsfront zeigten bemerkenswerte Überrepräsentation z. B. der „GO-terms“ „extrazelluläre Matrix“, Zellkommunikation“, „Antwort auf biotischen Stimulus“, Strukturmolekülaktivität“ und „Zellwachstum“. Marker der Aktivierung hepatischer Sternzellen überrepräsentiert in der invasionsfront, was die Durchführbarkeit einer Analyse differentieller Genexpression im genomweiten Rahmen anzeigt. Globale Genexpressionsstudien, auf den Tumorzellen in der in vitro Situation, in vivo und in der Invasionsfront zeigten insgesamt einen Anstieg zellulärer Spezialisierung von der in vitro zur Invasionsfront. Sezernierte proangiogenetische Chemokine zeigten eine Hochregulation in der Invasionsfront. Das beta catenin Gen war in der Invasionsfront 9.6 fach erhöht im Vergleich zur in vitro Situation. Die Überprüfung der transkriptionellen Aktivierung von beta catenin über die Prüfung der Promotoraktivität zeigte einen 18.4 fachen Anstieg in den Tumorzellen der Invasionsfront. Weiterhin war die Promotoraktivität (an Hand der Aktivität der mRNA des Alkalischen Phosphatase Reportergens) im Tumorinneren 3.5 fach höher als in der Zellkultur, was für einen transkriptionellen Mechanismus der beta catenin Regulation zusätzlich zu den posttranslationalen Mechanismen spricht. / Liver is most frequently populated by metastases and may therefore serve as a model organ for metastatic invasion. So the aim of this thesis is to understand the gene expression profiles and identify metastasis and invasion related genes. Differential gene expression was examined in three systems: A syngeneic mouse model, a xenograft model and five clinical specimens. Gene expression profiles of a syngenic mouse model and human clinical specimen revealed that the invasion front should be considered as a whole to find more overlapping potential target genes. Global gene expression studies on the host part of the invasion front, revealed a pronounced overrepresentation of GO-terms (e.g. “extracellular matrix”, “cell communication”, “response to biotic stimulus”, “structural molecule activity” and “cell growth”). Hepatic stellate cell activation markers were over-represented in the invasion front demonstrating the feasibility of a differential gene expression approach on a genome wide scale. Global gene expression studies of the tumor cells in vitro, in vivo and tumor part of the invasion front revealed an overall increase of cellular specialization from in vitro to the invasion front. Secreted angiogenic cytokines were found to be up regulated in the invasion front. Beta catenin gene of “cell adhesion” GO term was elevated 9.6 fold in invasion front compared to in vitro. Evaluation of transcriptional up-regulation of beta catenin by promoter activity showed an 18.4 fold increase in the tumor cells of the invasion front as compared to those from the faraway tumor. Promoter activity assessed by soluble human placental alkaline phosphatase reporter gene mRNA was 3.5 fold higher in the inner parts of the tumor than in vitro cells indicating a transcriptional mechanism of beta catenin regulation in addition to the posttranslational regulatory mechanisms.