Richard, Magali et al. (2009) International Worm Meeting "A visual screen for abnormal localization of cholinergic receptors tagged using homologous recombination."

Robert, Valerie, Duboin, Aurelien, Bessereau, Jean-Louis, Richard, Magali, Studer, Vincent

[International Worm Meeting, 2009]

Ionotropic acetylcholine receptors (AChRs) are pentameric ligand-gated ion channels that have been conserved throughout evolution. At the C. elegans neuromuscular junctions (NMJ) two types of AChRs mediate excitatory neurotransmission: N-AChRs, likely homomeric channels composed of the ACR-16 subunits, which are sensitive to nicotine, and L-AChRs, heteropentameric receptors activated by the drug levamisole. Several components involved in L-AChR formation and function have been identifed in the past using forward screens for mutants with decreased sensitivity to levamisole. However, resistance to levamisole is quite an indirect index of the numerous molecular mechanisms controling AChR composition, assembly, trafficking and localization. To study the biology of L-AChRs, we propose a novel approach based on the in vivo vizualization of tagged AChRs. Using the MosTIC technique recently developped in our laboratory, we have introduced, by homologous recombination, different fluorescent tags into the endogenous locus unc-29, which encodes one L-AChR subunit. We then used two different approaches to inactivate gene function (i) a chemical mutagenesis using EMS (ethylmethane sulfonate), (ii) RNA interference (RNAi) targeting 1300 genes selected for putative role in synaptic function. First, we will present mutants isolated in the EMS screen for AChRs clusters disappearance at the nerve ring. Second, we will describe a RNAi screen, for abnormal localization of AChRs at the NMJ, and the characterization of some putative candidates in which AChRs clusters seem to be altered. Third, we will demonstrate how microfluidic devices can improve the efficiency of visual screens.

Blaxter, Mark et al. (2015) International Worm Meeting "The Caenorhabditis Genomes Project."

Koutsovoulos, Georgios, Chandrasekar, Sinduja, Stevens, Lewis, Blaxter, Mark

[International Worm Meeting, 2015]

The sequencing of the genome of Caenorhabditis elegans remains one of the milestones of modern biology, and this genome sequence is the essential backdrop to a vast body of work on this key model organism. "Nothing in biology makes sense except in the light of evolution" (Dobzhansky) and thus it is clear that complete understanding of C. elegans will only be achieved when it is placed in an evolutionary context. While several additional Caenorhabditis genomes have been published or made available, a recent surge in the number of available species in culture makes the determination of the genomes of all the species in the genus a timely and rewarding project.We have initiated the Caenorhabditis Genomes Project. From material supplied by collaborators we have so far generated raw Illumina short-insert data for sixteen species. Where possible we have also generated mixed stage stranded RNASeq data for annotation. The data are being made publicly available as early as possible (warts-and-all) through a dedicated genome website at htttp://caenorhabditis.bio.ed.ac.uk, and completed genomes and annotations will be deposited in WormBase as mature assemblies emerge. We welcome additional collaborators to the CGP, whether to assemble new genomes or to delve into the evolutionary history of favourite gene sets and systems.Species sequenced thus far in Edinburgh: Caenorhabditis afra, Caenorhabditis castelli, Caenorhabditis doughertyi, Caenorhabditis guadeloupensis, Caenorhabditis macrosperma, Caenorhabditis nouraguensis, Caenorhabditis plicata, Caenorhabditis virilis, Caenorhabditis wallacei, Caenorhabditis sp. 1, Caenorhabditis sp. 5, Caenorhabditis sp. 21, Caenorhabditis sp. 26, Caenorhabditis sp. 31, Caenorhabditis sp. 32, Caenorhabditis sp. 38, Caenorhabditis sp. 39, Caenorhabditis sp. 40, Caenorhabditis sp. 43.[Samples have been supplied by Aurelien Richaud, Marie-Anne Felix, Christian Braendle, Michael Alion, Piero Lamelza].