19-P025 Bone dynamics during adult zebrafish caudal fin regeneration

19-P025 Bone dynamics during adult zebrafish caudal fin regeneration
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  changesintemporalandspatialexpressionpatternsofmarkersinregenerating and homeostatic worms. We aim to investigatewhether the genes may play a role in (re)programming positionalinformation during regeneration in  Schmidtea mediterranea .doi:10.1016/j.mod.2009.06.810 19-P023Anterior regeneration in the planarian  Schmidtea mediterranea: Revisiting the works of T.H. Morgan and C.M. Child Deborah Evans, Aziz Aboobaker University of Nottingham, Nottingham, United Kingdom Planaria possess a population of pluripotent adult stem cellscalled neoblasts, capable of differentiating into all cell typesrequired to regenerate entire animals from almost any fragment.The regeneration of different fragments is catalogued in a body of classical experiments by many eminent scientists. Here, weendeavour to explore these classical observations and insightsand combine them with molecular approaches in the planarian Schmidtea mediterranea .Morgandemonstratedthattheearlyanteriorblastemadisplaysthe potential to produce more than one head if divided laterally,while later in regeneration this plasticity is lost. Using the samenon-molecular criteria as Morgan, we find that in  S. mediterranea theanteriorblastemaiscommittedtoformonlyoneheadimmedi-ately after anterior regeneration is initiated. However, the timetakentoregulateregenerationandbeginproducingmissinglateralstructures increases with the length of time anterior regenerationhas proceeded before the division. We have developed the spatio-temporal characterisation of this process as an assay to discoversubtleRNAiregenerationphenotypesthatareaffectedbythispro-cess, but ultimately result in otherwise normal regeneration.Child observed a delay in regeneration in anterior blastemasinduced in more posterior positions. We have used a series of molecular markers to deduce that this difference may beobserved as early as 24 h of regeneration. We investigate the pos-sibility that there is a delay in specifying the fate of anterior blas-temas produced from more posterior regions and explore the roleof the hypothesised pre-existing Wnt signalling gradient in thisphenomenon.doi:10.1016/j.mod.2009.06.811 19-P024Studying regeneration in an emerging model crustacean Nikolaos Konstantinides, Michalis Averof  Institute of Molecular Biology and Biotechnology (IMBB), Iraklio, Crete,Greece Information on regeneration derive either from a small num-ber of model organisms with restricted regenerative capacity, orfrom non-model species that are difficult to access genetically(e.g. axolotls). The amphipod crustacean  Parhyale hawaiensis  isemerging as a promising model for studies in regeneration, as itis capable of regenerating all its appendages. Moreover, experi-mental embryology, lineage tracing, gene knockdown, overex-pression and transgenesis approaches on  Parhyale  are alreadyestablished. We are using mosaic analysis, marking different pop-ulations of cells with transgenic fluorescence markers, to dis-cover the progenitors of regenerating tissues and to elucidatetheir developmental plasticity.doi:10.1016/j.mod.2009.06.812 19-P025Bone dynamics during adult zebrafish caudal fin regeneration Sara Sousa, Nuno Afonso, Mariana Fonseca, Anto´nio Jacinto Instituto de Medicina Molecular (IMM), Lisbon, Portugal Some species such as zebrafish can regenerate several of itsorgans. In particular, the caudal fin can fully regenerate all its tis-sues, including bone. Behind this remarkable regenerative capac-ity is the ability to form a specialised tissue, the blastema. Wepropose to analyse the contribution of bone tissues to blastemaformation and to investigate how the blastema gives rise to newbone. We have started by analysing gene expression to try touncover the cellular and molecular identity of the tissue thatearly after amputation drives skeletal regeneration. For this char-acterization we are using several markers usually associated withbone-forming cells in mammals like the transcription factor generunx2 and makers for maturing osteoblasts: collagen type X andosterix. Furthermore we are characterizing the expression of car-tilage markers such as sox9a and collagen type II in the regener-ating bony rays upon amputation.To monitor bone dynamics during zebrafish caudal regenera-tion, we have improved imaging methodologies for long-termobservation of adult living zebrafish. We are using fast scanning confocal microscopy that allows scanning of a largevolume of tis-sue in a short period with minimal phototoxicity. This is coupledto an anesthetic apparatus that can maintain the fish still, aliveand regenerating for several hours. After imaging, the fish arefully recovered and can be imaged again several times a day mak-ing it possible to follow cells for many days. We are plan to useavailable bone-specific transgenic lines to study the dynamics of bone formation during fin regeneration.doi:10.1016/j.mod.2009.06.813 19-P026Zebrafish as a model to study the role of nerves in regeneratingappendages’ Mariana Simo ˜ es, Anto´nio Jacinto Instituo Medicina Molecular, Lisboa, Portugal It has been known for a long time that nerves are crucial forthe regenerative process of vertebrate appendages. Studies in fishand amphibians have shown that upon nerve fibre removal,regeneration does not occur properly. Axons are thought torelease mitogenic growth factors (MGFs) into the amputated S298  M E C H A N I S M S O F D E V E L O P M E N T  126 (2009) S291  –  S304
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