Analysis and Quantification of Functional Regeneration of Dendrite and Axon of PVD Neuron After Laser Injury in Caenorhabditis elegans.

Abstract

Research into nervous system injuries and regeneration has emerged as a crucial field of study. In many cases such as trauma or stroke, both axons and dendrites are equally damaged; however, studying injury and repair mechanisms in both neurite processes (axons and dendrites) of the same neuron has been challenging. Additionally, correlating the behavioral aspects of neuronal injury with anatomical regeneration is important for a better understanding of the functional rewiring process. Here, we describe protocols for injuring the dendrites and the axon of the PVD neuron of C. elegans using a two-photon infrared (IR) femtosecond laser system, and subsequent imaging of injured neurites during the course of regeneration. Additionally, we describe the protocols for the behavioral study concerning the PVD neuron and their analysis, which can offer valuable insights. These assays can be implemented to assess the function of the pathways that play specific roles in dendrite vs. axon regeneration. Key features • Dendritic and axonal injury in PVD neurons using a two-photon microscope. • Behavioral analysis involving harsh touch response and proprioception following neurite injury. • Correlation of dendrite and axon regeneration events with behavioral parameters.