PNG-1, A Peptide: N-Glycanase Limits Axon Outgrowth and Branching in Caenorhabditis elegans

Nasrin Habibi-Babadi, Université D'Ottawa / University Of Ottawa, Université D'Ottawa / University Of Ottawa
2016
Assembly of neuronal networks with distinct patterns of connectivity during nervous system development involves the growth, extension and branching of axons and dendrites. Over the years genetic and biochemical studies in model organisms have contributed significantly in identifying mechanisms regulating axon growth and extension. However the molecular mechanisms underlying axon branching remain unclear. The egg-laying neuronal circuitry in C. elegans has proven to be a robust system for
more » ... ying and characterizing novel genes involved in neuronal morphology. This circuitry which mediates egg-laying behavior in nematodes is composed of two families of motorneurons, HSNs and VCs, which are among the most branched neurons in C. elegans. A genetic screen for axon branch defects in the egg-laying neurons identified png-1 to disrupt neuronal morphology including axon branching. png-1 encodes a Peptide: N-glycanase (PNGase), a conserved cytosolic enzyme that removes N-linked sugar moieties from glycoproteins. In this thesis I present my work characterizing and examining png-1 and its role in mediating axon branching. Mutations in png-1 resulted in excessive ectopic axon branching in the VC4 and VC5 egg-laying neurons as well as branching in the normally unbranched AVL and DVB neurons. Behavioral analysis in these mutants revealed defects in egg-laying behavior and mild in-utero egg retention phenotypes. Cellular characterization shows ubiquitous expression of png-1 in many tissues including vulva cells, muscles, gonads, and neurons. My analysis also shows that png-1 acts both cell-autonomously and cell non-autonomously from neurons and epithelial cells to restrict axon branching around the vulva. Using a candidate gene approach I identified a deletion allele of the DNA repair gene, rad-23, to display axon branching defects and interact with png-1 within a common pathway to regulate axon branching. Additionally, through a genetic modifier screen for enhancers and suppressors of VC4-5 branching defects in png-1, I ide [...]
doi:10.20381/ruor-5022 fatcat:txt6eated5fb7kpmpsuwdv7dwq