Abstract
Gangliosides, sialic acid‐containing glycosphingolipids, are ubiquitously expressed in all eukaryotic cells and are localized primarily in the plasma membrane. For a rat pheochromocytoma cell line, PC12, which has been used frequently as a model for investigating events leading to neuronal differentiation, it is generally thought that GM1 is a major ganglioside, based on reactivity with the probe cholera toxin B subunit (Ctxb). From a series of biochemical studies, however, it has been reported that no GM1 is expressed in PC12 cells. In this study, we have reevaluated GM1 expression and Ctxb reactivity in PC12 cells and a subcloned line, PC12D cells. Flow cytometric analysis with Ctxb revealed that about 30–50% of PC12 cells were reactive with Ctxb. However, a detailed biochemical analysis showed that PC12 cells express abundantly a different ganglioside, fucosyl‐GM1, instead of GM1, and the reactivity of Ctxb in the PC12 cells actually arose from its interaction with fucosyl‐GM1, which also interacts with this ligand. Because it has been claimed that amyloid‐β protein (Aβ) interacts with GM1 in PC12 cells to provide “seeding” for amyloid to accumulate, we further evaluated this possibility and found that Aβ is mostly likely interacting with fucosyl‐GM1 in this cell line. Our data thus suggest that a specific interaction may occur between Aβ and fucosyl‐GM1 for the accumulation of amyloid in PC12 cells. © 2006 Wiley‐Liss, Inc.