Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4507

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

Esther T. Stoeckli

This protocol was adapted from "RNAi in Avian Embryos," contributed by Esther T. Stoeckli, Chapter 14, in RNAi: A Guide to Gene Silencing (ed. Hannon). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.


INTRODUCTION

This 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.


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Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos
Esther T. Stoeckli
Cold Spring Harb Protoc 2006: 4506. [Abstract] [Full Text]