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Journal of Histochemistry and Cytochemistry, Vol. 49, 285-292, March 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Immunochemical Characterization of a Chicken Egg Yolk Antibody to Secretory Forms of Rat Incisor Amelogenin

Giovanna Orsinia, Patrice Lavoiea, Charles E. Smithb, and Antonio Nancia
a Laboratory for the Study of Calcified Tissues and Biomaterials, Faculty of Dentistry, Université de Montréal, Montréal, Québec, Canada
b Division of Oral Biology, Faculty of Dentistry, McGill University, Montréal, Québec, Canada

Correspondence to: Antonio Nanci, Université de Montréal, Faculty of Dentistry/Stomatology, PO Box 6128, Station Centre-Ville, Montréal, QC, Canada H3C 3J7. E-mail: antonio.nanci@umontreal.ca

Amelogenins represent the major component of the organic matrix of enamel, and consist of several intact and degraded forms. A precise knowledge of their respective distributions throughout the enamel layer could provide some insight into their functions. To date, no antibody exists that can selectively detect the secretory forms of amelogenin. In this study we used the chicken egg yolk system to generate an antibody to recombinant mouse amelogenin. Immunoblots of whole homogenates from rat incisor enamel organs and enamel showed that the resulting antibody (M179y) recognized proteins corresponding to the five known secretory forms of rat amelogenin. Immunogold cytochemistry demonstrated that reactivity was restricted to ameloblasts and enamel. Secretory forms of amelogenin persisted in significant amounts throughout the enamel layer. The density of labeling was highest over the surface portion of the enamel layer, but enamel growth sites in this region showed a localized paucity of gold particles. Immunoreactivity was lowest over the mid-portion of the layer and increased moderately near the dentino–enamel junction. These results indicate that intact forms of amelogenin probably have a more complex distribution in the enamel layer than was heretofore suspected.

(J Histochem Cytochem 49:285–292, 2001)

Key Words: polyclonal antibody, amelogenin, secretory forms, immunoblotting, immunocytochemistry


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