Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos

INTRODUCTIONThis protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. The tissue is cut in slices using a vibratome or tissue slicer. The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis. Slices can also be used for tracing axon tracts with lipophilic dyes rather than immunohistochemical detection of all neurites with anti-neurofilament antibodies.