Distribution of antibody- and lectin-binding sites on dissociated blastomeres from mouse morulae: evidence for polarization at compaction

Abstract

The distribution of binding sites for rabbit anti-species antiserum, Concanavalin A (Con A) and peanut agglutinin (PNA) on dissociated blastomeres from 2- to 16-cell mouse embryos has been investigated using direct and indirect fluorescence techniques. With each ligand, paraformaldehyde-fixed blastomeres from 2- to 8-cell precompact embryos were uniformly surface labelled; the majority (77 %) of late compact 8-cell blastomeres showed quantitative polarization of surface lebelling; and 16-cell blastomeres were either polarized (53·3%) or uniformly surface labelled. Binding of fluorescein-conjugated PNA increased at the 16-cell stage. Labelling patterns on unfixed blastomeres were similar to those on fixed blastomeres except that surface label was patched and became internalized, most rapidly from the less heavily labelled areas of 8- and 16-cell blastomeres. Quantitative polarization of binding sites at postcompaction stages was detected after (i) fixation, (ii) pretreatment and labelling in the presence of azide, cytochalasin D and/or colcemid, or (iii) labelling with monovalent Fab_x antibody fragments. It is probably due, therefore, to the presence of microvilli at the heavily labelled pole, which increase surface area and are known to become localized to the outer surface of the compact morula (Ducibella, Ukena, Karnovsky & Anderson, 1977). The possibility that the cleavage of polarized blastomeres into dissimilar daughter blastomeres could provide a mechanism for the spatial differentiation of the inner cell mass and trophectoderm of the blastocyst is briefly discussed.