Elsevier

Brain Research

Volume 678, Issues 1–2, 24 April 1995, Pages 99-109
Brain Research

Developmentally-regulated lectin binding in the embryonic mouse telencephalon

https://doi.org/10.1016/0006-8993(95)00173-NGet rights and content

Abstract

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.

References (50)

  • LisH. et al.

    Biological properties of lectins

  • MargolisR.K. et al.

    Structural changes in brain glycoproteins during development

    Brain Res.

    (1974)
  • RathjenF.G. et al.

    Cholera toxin binding to cells of developing chick retina analyzed by fluorescence-activated cell sorting

    Dev. Brain Res.

    (1981)
  • SteindlerD.A. et al.

    Boundaries during normal and abnormal brain development: in vivo and in vitro studies of glia and glycoconjugates

    Exp. Neurol.

    (1990)
  • StempleD.L. et al.

    Isolation of a stem cell for neurons and glia from the mammalian neural crest

    Cell

    (1992)
  • SuchyS.F. et al.

    A monoclonal antibody, WCCC4, recognizes a developmentally regulated ganglioside containing α-fucose present in the rat nervous system

    Brain Res.

    (1988)
  • WillingerM. et al.

    GM1 ganglioside as a marker for neuronal differentiation in mouse cerebellum

    Dev. Biol.

    (1980)
  • AbneyE.R. et al.

    Astrocytes, ependymal cells and oligodendrocytes develop on schedule in dissociated cell cultures of embryonic rat brain

    Dev. Biol.

    (1984)
  • AchesonA. et al.

    NCAM polysialic acid can regulate both cell-cell and cell-substrate interactions

    J. Cell Biol.

    (1991)
  • AngevineJ.B. et al.

    Autoradiographic study of cell migration during histogenesis of cerebral cortex in the mouse

    Nature

    (1961)
  • AsouH. et al.

    Cellular localization of GM1-ganglioside with biotinylated choleragen and avidin peroxidase in primary cultured cells from rat brain

    J. Histochem. Cytochem.

    (1983)
  • BarbinG. et al.

    Brain astrocytes express region-specific surface glycoproteins in culture

    Glia

    (1988)
  • Blass-KampmannS. et al.

    In vitro differentiation of E-N-CAM expressing rat neural precursor cells isolated by FACS during prenatal development

    J. Neurosci. Res.

    (1994)
  • BottensteinJ.E. et al.

    Growth of a rat neuroblastoma cell line in serum-free supplemented medium

  • The Boulder Committee

    Embryonic vertebrate central nervous system: revised terminology

    Anat. Rec.

    (1970)
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