Changes in lectin-mediated agglutinability during primary embryonic induction in the amphibian embryo

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Abstract
The present study was undertaken to investigate structural alterations at the surface of presumptive neural cells after primary embryonic induction. For this purpose, plant lectin-mediated agglutinability of dissociated cells from the epiblast of Bufo arenarum gastrulae was tested. Two fragments of epiblast were excised from the same mid-gastrula: one from the dorsal side of the egg, making contact with the invaginating chordamesoblast and assumed to be composed of determined cells and the other from the ventral region of the egg, facing the blastocoele cavity and assumed to be composed of undetermined cells. Cells of the pooled fragments were dissociated in calcium-free Holtfreter’s solution with potassium oxalate and incubated in the presence of different concentrations of phytohemagglutinin and concanavalin A. Epiblast cells overlying the archenteron roof are less agglutinated with both lectins than undetermined cells. On the other hand, when egg fragments were removed from the dorsal and ventral regions of early gastrulae before the archenteron was formed, no significant difference in lectin-mediated agglutinability was observed, even after having been cultured in vitro in absence of inducing tissue. These results suggest that the target of the inducing signal generated in the mesoblast is likely to be located on the surface of epiblast cells. Additional experiments showed that cells pretreated with colchicine, cytochalasin B or colchicine and cytochalasin B simultaneously exhibit no significant variation in agglutinability, suggesting that the cytoskeleton was not be involved in the cell surface alteration here described. Treatment of whole embryos or sandwich explants with concanavalin A or phytohemagglutinin has no effect on neural tube formation, suggesting that the carbohydrate-containing binding sites for these lectins are not involved in primary embryonic induction. Changes in cell agglutinability described in this paper are to be interpreted thus as a secondary expression of structural alterations in the cell surface concomitant with neural determination.