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Journal of Histochemistry and Cytochemistry, Vol. 47, 1189-1200, September 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Ultrastructural Localization of Proteins Involved in Sea Urchin Biomineralization

Laurent Ameyea, René Hermannb, Christopher Killianc, Fred Wiltc, and Philippe Duboisa
a Laboratory of Marine Biology, Free University of Brussels, Brussels, Belgium,
b Laboratory for Electron Microscopy 1, Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
c Department of Molecular Cell Biology, University of California, Berkeley, California

Correspondence to: Laurent Ameye, Laboratoire de Biologie Marine, CP 160 / 15, Université Libre de Bruxelles, 50 Av. F.D. Roosevelt, 1050 Bruxelles, Belgium.

Three skeletal tissues of the adult echinoid Paracentrotus lividus (the pedicellaria primordium, the test, and the tooth) were immunolabeled with three sera raised against the total mineralization organic matrix and two specific matrix proteins (SM30 and SM50) from the embryo of the echinoid Strongylocentrotus purpuratus. Two conventional chemical fixation protocols and two high-pressure freezing/freeze–substitution protocols were tested. One conventional protocol is recommended for its good preservation of the ultrastructure, and one high-pressure freezing/freeze–substitution protocol is recommended for its good retention of antigenicity. Immunolabeling was obtained in the three adult tissues. It was confined to the active skeleton-forming cells and to the structured organic matrix. The results indicate that the matrix proteins follow the classical routes of secretory protein assembly and export and suggest that SM30 and SM50 are a part of the tridimensional network formed by the organic matrix before the onset of mineralization. They show that the genetic program of part of skeletogenesis is conserved among different calcification models and developmental stages. (J Histochem Cytochem 47:1189–1200, 1999)

Key Words: biomineralization, sea urchin, immunocytolabeling, organic matrix, tooth, skeleton, high-pressure freezing, freeze–substitution


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