Ciliated Sensory Neuron Defects in Caenorhabditis elegans
Presented here is research investigating genes that are involved in the development and maintenance of ciliated nerve endings in the nematode Caenorhabditis elegans. C. elegans utilizes a subset of neurons, referred to as ciliated sensory neurons, to sense certain changes in its environment. There are two amphid sensilla (sense organs) that mediate exposure of these ciliated endings to the animal's external environment. Those ciliated endings that penetrate the cuticle are responsible for a myriad of behaviors that range from chemotaxis to osmotic avoidance, but in general function for the reception of environmental cues and stimuli. The intraflagellar transport (IFT) process facilitates the morphogenesis of these ciliated endings, and animals lacking intact ciliated endings may not be able to detect nourishment, hazardous environments, or other worms for mating. Mutant strains used in this study were generated by EMS mutagenesis of wild-type N2 animals and a subsequent screen of those worms displaying significant cilia dysfunction as evidenced by their dye-filling defective (Dyf) phenotype. Cilia-mediated uptake of lipophilic DiI into six pairs of amphid sensory neurons and two pairs of phasmid sensory neurons is expected in wild-type (N2) animals, but in Dyf animals, this dye-filling is disrupted, either through morphological defects, or deleterious mutations in the IFT process. To investigate the morphogenesis of cilia in C. elegans, we analyzed two specific mutant strains, WX737 dyf-3(og022)IV and PK841 dyf-15(pk841)V, that are defective in the uptake of fluorescent dye DiI and abnormal in sensory cilium structure. Through a variety of genetic mapping techniques, we were able to successfully map experimental gene dyf-15(pk841) to an interval of 2.84cM on chromosome V, and identify og022 as an allele of the gene dyf-3. It has been previously shown that dyf-3 expression is detected in 26 chemosensory neurons, including six IL2 neurons, eight pairs of amphid neurons (ASE, ADF, ASG, ASH, ASI, ASJ, ASK and ADL) and two pairs of phasmid neurons (PHA and PHB). Analysis of cilium malformation and the presence of a recognition sequence for the DAF-19 transcription factor suggest that dyf-3 is involved in the intraflagellar transport system complex B.