Immunohistochemical Localization of Tubulin in the Somite of the Early Chick Embryo

Abstract

In this experiment I attempted to devise a procedure for the localization of the cytoskeletal protein tubulin in the somite of the early chick embryo. I also checked the distribution of tubulin in the early chick somite for any possible relationship to the epithelial-mesenchymal transition that occurs in these somitic ceils. Newly fertilized chicken eggs were incubated for approximately 48 hr; then the embryos were removed from the egg, fixed, embedded, and frozen for cryostat sectioning. After sectioning the embryos, I attempted to localize tubulin by attaching a primary antibody to the tubulin molecule followed by a secondary antibody and an enzyme (peroxidase). A peroxidase substrate was then introduced to the tissue, and the enzyme reacted with it to produce a colored reaction product which served as the indicator for the presence and distribution of the tubulin.