Temporal changes in lectin binding of peri-implantation mouse blastocysts |
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Authors: | Daniel J Chavez Allen C Enders |
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Institution: | Department of Human Anatomy, School of Medicine, University of California, Davis, California 95616 USA |
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Abstract: | Mouse blastocysts were exposed to a series of ferritin-conjugated lectins during Day 5 (preadhesive) and Day 6 (adhesive; collected Day 5, 24 hr in vitro) of embryogenesis to determine whether there were any changes in lectin binding characteristics that coincided with the acquisition of adhesiveness. After exposure to lectin, the blastocysts were processed for electron microscopy and lectin binding sites were determined by visualization of ferritin particles with the electron microscope. No binding sites were observed for either Dolichos biflorus agglutinin or soybean agglutinin on blastocysts from either stage examined. Binding sites for Ulex europaeus agglutinin, Con A, and wheat germ agglutinin were seen on blastocysts from both stages without apparent increase or reduction in binding sites from either stage. Ricinus communis agglutinin-I (RCA-I) bound heavily to the surface of Day 5 blastocysts and did not bind at all to Day 6 blastocysts and did bind, though with apparent diminution, to Day 6 blastocysts, as compared with the binding observed on Day 5 blastocysts. Peanut agglutinin (PNA) did not bind at all to Day 5 blastocysts but did bind heavily to the surface of Day 6 blastocysts. Both RCA-I and PNA bound to the surface of embryos during Day 5 of delayed implantation, thus indicating that neither the appearance of PNA binding sites on Day 6 blastocysts nor the apparent reduction of RCA-I binding sites on Day 6 blastocysts could be solely implicated in the acquisition of adhesiveness. PNA binding sites were abolished from the surface of Day 6 blastocysts by treatment with Pronase, indicating that the PNA binding molecule was associated with a glycoprotein rather than a glycolipid. |
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Keywords: | anti-R goat anti-rabbit IgG CSP cell surface protein DAB diaminobenzidine ECM extracellular matrix Fab the antibody-binding fragment of IgG Fc the non-antibody binding fragment of IgG Fe-anti-R ferritin-conjugated goat anti-rabbit IgG GAG glycosaminoglycans IgG immunoglobulin G LETS large external transformation-sensitive protein PAP peroxidase anti-peroxidase PBS phosphate-buffered saline (pH 7 2) Ranti-FN rabbit antifibronectin antiserum RR ruthenium red SDS sodium dodecyl sulfate SFA fibroblast surface antigen TBS Tris-buffered saline |
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