Résumé :
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Communication n° 58. We study muscle degeneration due to the absence of dystrophin. This protein and other molecules of the dystrophin complex have conserved homologues in the nematode Caenorhabditis elegans genome. The structural and molecular similarity between human and C.elegans muscles allows us to use this nematode as a relevant model. We are now using C.elegans as a tool to look for genes that would have an effect on muscle degeneration. In order to do so, we are using the RNA interference (RNAi) technique which allows the post-transcriptional inactivation of a chosen gene. We are performing these tests on nematodes presenting a dystrophin- and time-dependent muscle degeneration (see poster by Carre-Pierrat et.al). To speed up the RNAi screen, we try to automate the different steps of the protocol. We quantify muscle degeneration by the number of muscular cell missing in the nematode. It has been counted so far manually under the microscope. To allow automatic counting of the cells, we use a strain in which muscle nuclei have been made fluorescent by a gfp transgene. A missing cell is represented by a missing dot in the fluorescent pattern. This pattern is analysed by a program that calculates several statistical and descriptive parameters for each worm and for an entire population of worms eliminating unwanted worms (young, sick,…). All these parameters are then presented as tables and graphs to help decide whether the muscular state of the nematodes population has improved or not. Finally, data are recorded into a database in which it is also possible to have information about the inactivated gene.
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