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Journal of Histochemistry and Cytochemistry, Vol. 50, 617-628, May 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Localization of Caveolin 1 in Aortic Valve Endothelial Cells Using Antigen Retrieval

Nalini M. Rajamannana, Margaret J. Springettc, Larry G. Pedersonb, and Stephen W. Carmichaeld
a Department of Cardiology and Cardiovascular Diseases, Mayo Foundation and Clinic, Rochester, Minnesota
b Department of Biochemistry and Molecular Biology, Mayo Foundation and Clinic, Rochester, Minnesota
c Electron Microscopy Core Facility, Mayo Foundation and Clinic, Rochester, Minnesota
d Department of Anatomy, Mayo Foundation and Clinic, Rochester, Minnesota

Correspondence to: Nalini M. Rajamannan, Northwestern U. Medical School, 675 N St Clair St. Suite 10-240, Chicago, IL 60611. E-mail: n-rajamannan@northwestern.edu

Ultrastructural analysis of aortic valve endothelial cells subjected to growth arrest revealed many vesicles defined as caveolae by the localization of caveolin. Translocation of caveolin after exposure to oxidized LDL suggests that the localization of caveolin may be a valuable tool to study models of early atherogenesis. In this study, several antigen retrieval protocols were tested in osmium-fixed and Spurr-embedded cells to determine the optimal method of antigen retrieval in our model system. SDS produced the most consistent labeling pattern. A quantitative evaluation revealed that SDS significantly increased the labeling density in Spurr-embedded cells. The labeling pattern appeared as clusters of gold particles, 15–40 nm in diameter, that were associated with membranes of a similar size which may represent the neck region of the caveolae.

(J Histochem Cytochem 50:617–627, 2002)

Key Words: antigen retrieval, caveolin 1, immunoelectron microscopy, aortic valve endothelial cells, caveolae, quantitation, plasmalemmal vesicles, gold labeling, ultrastructure, LR White, Spurr resin


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