Implementing A Cell Culture Method for Imaging C. elegans Neurons and HighThroughput SNIP-SNP Mapping of C. elegans Suppressor Mutations
Fiona Gowen
Project Advisor: Renee Baran

     The imaging of microtubules using GFP-tagged proteins can provide information about microtubule-mediated mechanisms in neurons, such as axon outgrowth synapse formation, organelle and vesicle biogenesis, and neuron transport. GFP markers expressed by individual embryonic cells allow the identification of these neurons for imaging in vivo under epi-fluorescent microscopy and enables collection of cells through Fluorescent-Activated Cell Sorting (FACS) and mass cell culture to image cells in vitro. Although cells grown in vivo have been taken out of their growth environment, some cell processes such as microtubule activity can be viewed more easily in individual cells grown in vitro.  To isolate a specific subset of neurons, the D motor neurons, we used C. elegans strains that express either Green Fluorescent Protein (GFP) or Red Fluorescent Protein (RFP) transgenes in these cells. Mixtures of neuronal and non-neuronal cells were successfully isolated and cultured.
     A second ongoing project is the mapping and cloning of C. elegans suppressor mutations. The location of a mutation can be mapped to a specific chromosomal location using SNIP-SNP mapping. SNPs (Single Nucleotide Polymorphisms) are areas of a chromosome possessing a point mutation. SNIP-SNPs are SNPs in which the point mutation results in a change in a restriction enzyme site that can be detected by PCR and gel electrophoresis. We used this method to map the stub-3 (suppressor of tubulin mutant defects) to the right arm of C. elegans chromosome 3.

Support provided by: Howard Hughes Medical Institute Undergraduate Science Education Grant