The role of dpy-11 within C. elegans in maintaining organ morphogenesis and observable reversion in dpy-11 mutants, Bhatt, Asha; Foster, Andrea
College of Science and Mathematics
Dpy-11, a gene found in Caenorhabditis elegans, encodes a membrane associated thioredoxin- like protein (TRX) that influences body shape and morphogenesis. The dpy-11 protein is solely expressed in the hypodermis and is essential in producing a normal body form in C. elegans. The TRX protein produced by C. elegans is homologous to the human TMX4 protein which is up-regulated in melanoma cells. The knockdown of the human TMX4 protein can be used as a cancer therapy to down-regulate TMX4 in melanoma cells. When the dpy-11 is knocked-down, these nematodes appear to have a dumpy phenotype which is observed by their short and plump characteristics. RNAi was used to induce the knockdown of the dpy-11 gene. A vector containing the RNAi insert targeting the dpy-11 gene as well as the ampicillin resistance gene was obtained from the DNA Dolan Learning Center at Cold Spring Harbor Laboratories. Escherichia coli were transformed with this vector, and then fed to nematodes to induce the knockdown of the dpy-11 gene. The resulting dumpy phenotype, specifically the area of the worm, was measured and compared to wild-type C. elegans through a computer program, Image J. Furthermore the concentrations of IPTG were manipulated to determine its effect on the dumpy phenotype. IPTG induces the promoter and therefore we expect that at higher concentrations of IPTG the worms will be more dumpy. Taurine, a dietary supplement, reduces ER stress and was used to assess reversion of the dpy-11 mutants. Nitric oxide promotes longevity of C. elegans and was therefore used to evaluate survival rate of mutants compared to wild-type C. elegans. Both taurine and nitric oxide were used by dissolving them in the NGM agar and using the RNAi methods mentioned above. We expect the dpy-11 mutants to revert back to their wild-type state with taurine in the NGM agar. We also expect the survival rate of the mutants to increase with the addition of nitric oxide to the NGM agar. Results will be presented at the symposium.
