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EVOLUTION AND DEVELOPMENT Élio Sucena Principal Investigator PhD in Evolution and Development, Genetics, Cambridge, UK, 2001 Post-Doc at Princeton University, USA Post-Doc at University of Western Ontario, Canada Liaison with the Champalimaud Foundation Principal Investigator at the IGC since 2003 Research in my lab focuses on evolutionary novelties, defined as a novel body part that is neither homologous to any body part in the ancestral lineage nor serially homologous to any other body part of the same organism. This concept can be extended to other traits, for example, physiological or behavioural. We have identified several instances of novelty, as defined above, that we are experimentally dissecting: 1. At the genetic level, looking at gene function evolution upon gene duplication; 2. At the cellular level, approaching immune cell function diversity in Drosophila; 3. At the morphological level by studying the evolutionary origin of dorsal appendage formation in the Drosophila clade. DISSECTING THE DEVELOPMENTAL GENETIC BASIS OF EVOLUTIONARY NOVELTY The eggshell's dorsal appendages are structures unique to the Drosophila lineage, found throughout the lineage in various numbers, shapes and sizes. With Adrien Fauré and Claudine Chaouiya we have progressed in the establishment of a model that recapitulates the epithelial patterning, and with which we are now able to explore the variation in number this structure displays across Drosophila species. In addition, our research into eggshell patterning of Ceratitis capitata, a species without dorsal appendages outside of the Drosophila clade, has led to a renewed understanding of the role played by the Dpp pathway during oogenesis. With the combination of these two lines of research we continue to investigate both the possible origin as well as the diversification of this evolutionary novelty. GROUP MEMBERS Nelson Martins (Post-doc) Kohtaro Tanaka (Post-doc, started in May) Alexandre Leitão (PhD student) Barbara Vreede (PhD student) Vítor Faria (Research Assistant) COLLABORATORS Jen Sheen (Massachusetts General Hospital, USA) Mark Borowsky (Massachusetts General Hospital, USA) Brad Chapman (Massachusetts General Hospital, USA) Ignacio Rubio Somoza, (Max Planck Institute for Developmental Biology, Germany) Detlef Weigel (Max Planck Institute for Developmental Biology, Germany) Pedro L. Rodriguez Egea (Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia, Spain) Markus Teige (Max F. Perutz Laboratories, Univ. Vienna, Austria) Andreas Bachmair (Max F. Perutz Laboratories, Univ. Vienna, Austria) Claudine Chaouiya (IGC, Portugal) Adrien Fauré (IGC, Portugal) Thiago Carvalho (IGC, Portugal) Jocelyne Demengeot (IGC, Portugal) Luis Teixeira (IGC, Portugal) Fernando Roch (Centre de Biologie et Developpement, France) Sara Magalhães (Faculdade de Ciências, Universidade de Lisboa, Portugal) FUNDING Fundação para a Ciência e a Tecnologia (FCT), Portugal Instituto Gulbenkian de Ciência (IGC), Portugal Comparing the patterning network of Drosophila with a species without dorsal appendages, the mediterranean fruitfly Ceratitis capitata, has pointed us toward a key node in the network: the gene mirror. This transcription factor is absent in Ceratitis oogenesis, but performs a key function in defining the cells that make up the dorsal appendages, as well as defining the dorso-ventral axis of the future Drosophila embryo. We are currently exploring this co-option in more detail. FUNCTIONAL EVOLUTION UPON GENE DUPLICATION We have been studying a recently expanded gene family in Drosophila, the Three Finger Domain Protein (TFDP), in particular the nine genes of clusters III and V. Given their expression patterns in the Drosophila eye disc, we hypothesise that, in higher Diptera, TFDP genes have been co-opted by the ancestral eye development gene network. Different genes of this recently expanded family have been deployed in specific glial/neural cell subsets as to participate in the highly dynamic process leading to the formation of both photoreceptor axonal tracks and their respective myelin sheaths. This study establishes the TFDP gene family evolution as an example of neo/sub-functionalisation by gene duplication. Further work will clarify the specific impact of regulatory evolution over the gene network driving the cellular mechanisms in the developing eye of holometabolous insects. In order to have a comprehensive description of expression pattern evolution of these nine TFDP genes in Drosophila, we extended our analysis to Tribolium, Anopheles, Megaselia and Ceratitis. With this wide phylogenetic coverage, we have a clear idea of the duplication history of these genes and hence a strong basis to define instances of sub- and neo-functionalisation for future promoter dissection. IMMUNE CELL FUNCTION DIVERSITY IN DROSOPHILA We had previously shown that, contrary to the current view, Drosophila plasmatocytes (the functional analogues of vertebrate macrophages) constitute a heterogeneous population. For this we have established a novel protocol for PARASITOID WASP EGG SURROUNDED BY HOST IMMUNE CELLS (PLASMATOCYTES). Heterogeneity of plasmatocyte population is apparent by the differential expression of green (Cg25C) and red fluorescence (eater). IGC ANNUAL REPORT ‘11 RESEARCH GROUPS 62
hemocyte purification using a combination of GFP lines, specific staining and FACS analysis. Recently, we have shown that immune challenge leads to changes in total cell number and subset proportions. Moreover, using G-trace technique, we have determined that such changes are not due to shifts in cell identity but rather the result of differential proliferation and/or recruitment. We have now found new plasmatocyte subsets and shown that some of these subsets change dynamically during larval development. After confirming the long standing hypothesis that hemocytes are essential to resist parasitoid egg infections we have described the plasmatocyte heterogeneity during this immune response. The results show that there is a dynamic change in all plasmatocyte subsets during the immune response while they maintain the expression of a differentiated plasmatocyte marker. EVOLUTION OF Drosophila melanogaster IMMUNE RESPONSE The mechanistic basis of the immune response in Drosophila has been widely studied; however, little is known about the evolutionary and physiological mechanisms that drive local adaptation to pathogens. It is also unknown if this adaptation is dependent on factors such as the infected life-stage, route of infection, pathogen type or multiple infections, and the associated trade-offs. Using experimental evolution in Drosophila, to different and contrasting immune challenges, we have established the grounds to address these questions and gain insights about the genes and mechanisms involved in adaption to pathogens. We have imposed 20 generations of experimental evolution in Drosophila under different immune challenges. These replicate populations have adapted to contrasting conditions and perform significantly better that controls. We are currently testing for the establishment of trade-offs and will pursue the genetic basis of this adaptation. Cages containing Drosophila populations undergoing experimental evolution. Phylogenetic relationships between different dipterans used in the lab to study the origin and diversification of egg dorsal appendages (right panels). IGC ANNUAL REPORT ‘11 RESEARCH GROUPS 63
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The Instituto Gulbenkian de Ciênci
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ORGANISATION CALOUSTE GULBENKIAN FO
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established a unique and diverse hu
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Page 81 and 82: IMMUNE REGULATION Carlos E. Tadokor
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António Coutinho Professional Meri
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GRADUATE TRAINING AND EDUCATION
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Cells to organisms I 24 - 28 Octobe
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THESES MSc THESES Sofia Pereira Dev
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On the origin and diversification o
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HELENA COSTA Mechanism of IL-8 indu
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The centromere: A showcase for epig
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PUBLIC ENGAGEMENT IN SCIENCE
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Science Projects Support Programme
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FUNDRAISING Maria João Leão Head
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PARTNERS AND SPONSORS Several partn
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