Developmentally-regulated lectin binding in the embryonic mouse telencephalon

Cell-surface carbohydrate epitopes are important determinants in cell-cell and cell-matrix interactions, and oligosaccharide groups are structural components of many growth factor receptors and cell adhesion molecules. These epitopes may participate in the regulation of stem cell proliferation and differentiation during central nervous system development. To further understand these cellular phenomena, it is important to define the changes in neuroepithelial cell-surface carbohydrate expression during development. We used a panel of fluorescein-conjugated lectins to label live, freshly dissociated cells from the embryonic day 11 to 18 (E11 to E18) mouse telencephalon. The intensity and heterogeneity of lectin labeling was assessed by flow cytometry. The lectins that we examined exhibited widely varying levels of labeling intensity. Lectins with the highest degree of binding included cholera toxin B subunit (CTB), which binds primarily to the gangliosides GM1 and GD1b, phaseolus vulgaris erythroagglutinating lectin (PHA-E), which binds to a variety of cell adhesion molecules, and wheat germ agglutinin (WGA). Many lectins showed increasing labeling intensity and cellular heterogeneity as development progressed. To determine if the observed cellular heterogeneity in lectin binding reflected biological differences in neuroepithelial cell subpopulations, cells from the E14 telencephalon were separated into two populations based on their intensity of CTB labeling using a fluorescence activated cell sorter. The population of weakly CTB labeled cells contained more than four times as many cells in S-phase of the cell cycle than the population of intensely CTB labeled cells. These observations suggest that lectin cytochemistry and flow cytometry can be useful in identifying specific cell subpopulations of neuroepithelial precursor cells during development, allowing their isolation and characterization in vitro.