Characterization of GD3 ganglioside as a novel biomarker of mouse neural stem cells

Abstract

Neural stem cells (NSCs) are undifferentiated neural cells characterized by their high proliferative potential and the capacity for self-renewal with retention of multipotency. Over the past two decades, there has been a huge effort to identify NSCs morphologically, genetically, and molecular biologically. It is still controversial, however, what bona fide NSCs are. To define and characterize NSCs more systematically, it is crucial to explore novel cell-surface marker molecules of NSCs. In this study, we focused on GD3, a b-series ganglioside that is enriched in the immature brain and the subventricular zone (SVZ) of the postnatal and adult brain, and evaluated the usefulness of GD3 as a cell-surface biomarker for identifying NSCs. We demonstrated that GD3 was expressed in more than 80% of NSCs prepared from embryonic, postnatal, and adult mouse brain tissue by the neurosphere culture method. The percentage of GD3-expressing NSCs in neurospheres was nearly the same as it was in neurospheres derived from embryonic, postnatal, and adult brains but decreased drastically to about 40% after differentiation. GD3(+) cells isolated from embryonic mouse striata, postnatal, and adult mouse SVZs by fluorescence-activated cell sorting with an R24 anti-GD3 monoclonal antibody efficiently generated neurospheres compared with GD3(-) cells. These cells possessed multipotency to differentiate into neurons, astrocytes, and oligodendrocytes. These data indicate that GD3 is a unique and powerful cell-surface biomarker to identify and isolate NSCs.

Sur8/Shoc2 Involves Both Inhibition of Differentiation and Maintenance of Self-Renewal of Neural Progenitor Cells via Modulation of Extracellular Signal-Regulated Kinase Signaling

“…NPCs retain stem cell characteristics, including a high proliferative potential and the ability to selfrenew [30]. In this study, we identified Sur8, the scaffold protein of the Ras-ERK pathway [6][7][8], as a factor that maintains self-renewal capacity and the undifferentiated state in NPCs.…”

Interaction of ganglioside GD3 with an EGF receptor sustains the self-renewal ability of mouse neural stem cells in vitro

“…In later developmental stages, however, more complex gangliosides prevail, particularly GM1 (Galβ1-3GalNAcβ1-4(NeuAcα2-3)Galβ1-4Glcβ1-1′Cer), GD1a (NeuAcα2-3Galβ1-3GalNAcβ1-4(NeuAcα2-3)Galβ1-4Glcβ1-1′ Cer), GD1b (Galβ1-3GalNAcβ1-4(NeuAcα2-8NeuAcα2-3) Galβ1-4Glcβ1-1′Cer), and GT1b (NeuAcα2-3Galβ1-3GalNAcβ1-4 (NeuAcα2-8NeuAcα2-3)Galβ1-4Glcβ1-1′Cer) (2,3). Because of the spatiotemporal expression patterns, gangliosides abundant in embryonic brain, such as GD3 and the c-series ganglioside antigen marker A2B5, have been considered to be useful stage-specific markers of early brain development (4,5). Mounting evidence supports the notion that gangliosides serve regulatory roles in neurogenesis through modulating processes, such as intercellular recognition, interaction, adhesion, reception, and/ or signaling (6)(7)(8)(9).…”

“…In primary NSCs prepared from mice, the expression of GD3 was found to coexist with that of stage-specific embryonic antigen 1, another mouse NSC marker, in cultured neurospheres and neuroepithelial cells (NECs), but not in differentiated cells. For that reason, GD3 can be considered a useful NSC surface marker (4). The specific expression of GD3 in NSCs and its dynamic change during neural development prompted us to investigate the functional role of GD3 in regulating NSC fate determination.…”

Lewis X-carrying N-Glycans Regulate the Proliferation of Mouse Embryonic Neural Stem Cells via the Notch Signaling Pathway

“…NSC Culture-NSCs were prepared in the form of neurospheres according to methods described previously (22,24). In brief, single cell suspensions prepared from the striata of ICR mouse embryos (embryonic day 14.5) were cultured in Neurobasal-A medium (Invitrogen) containing B27 serum-free supplement (Invitrogen), L-glutamine (Invitrogen), 20 ng/ml basic FGF (PeproTech, Rocky Hill, NJ), and 20 ng/ml of epidermal growth factor (EGF; PeproTech).…”