Department of Neurology, Fujita Health University School of Medicine, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake, Aichi 470-1192, Japan.
FASEB J. 2012 Jul;26(7):3065-74. doi: 10.1096/fj.11-198630. Epub 2012 Apr 16.
Mutations in the presenilin 1 (PS1) gene are associated with early onset familial Alzheimer's disease (FAD). In this study, we found that the expression of mutant-PS1 in stable transfectants of SH-SY5Y neuroblastoma cells results in a reduction of the biosynthesis and steady-state levels of glucosylceramide. As an in vivo corroboration of these data, there was a significant reduction of brain glucosylceramide and gangliosides in an animal model of FAD. In mutant-PS1-transfectants (I143T, G384A), immunocytochemistry disclosed a remarkable reduction of glucosylceramide synthase (GlcT-1)-like immunoreactivity in the cells when compared with those of mock- and wild-PS1 transfectants. Immunoprecipitation of GlcT-1 protein from mutant-PS1 transfectants demonstrated a marked reduction in GlcT-1 protein, but there was no reduction in the levels of GlcT-1 mRNA. Both coprecipitation and γ-secretase inhibition experiments suggest that mutant-PS1 seems to form a complex with GlcT-1 protein and to be involved in GlcT-1 degradation, which was never found in other cell types. Thus, mutations in the PS1 gene result in profound glycosphingolipids abnormalities by abnormal molecular interaction with GlcT-1.
早发型家族性阿尔茨海默病(FAD)与早老素 1(PS1)基因突变有关。在这项研究中,我们发现,突变型 PS1 在 SH-SY5Y 神经母细胞瘤细胞的稳定转染子中的表达导致葡糖神经酰胺的生物合成和稳态水平降低。作为这些数据的体内佐证,在 FAD 的动物模型中,脑葡糖神经酰胺和神经节苷脂显著减少。在突变型 PS1 转染子(I143T、G384A)中,与 mock 和野生型 PS1 转染子相比,免疫细胞化学显示细胞中的葡糖神经酰胺合酶(GlcT-1)样免疫反应性显著降低。从突变型 PS1 转染子中免疫沉淀 GlcT-1 蛋白表明 GlcT-1 蛋白显著减少,但 GlcT-1 mRNA 水平没有减少。共沉淀和γ-分泌酶抑制实验表明,突变型 PS1 似乎与 GlcT-1 蛋白形成复合物,并参与 GlcT-1 降解,这在其他细胞类型中从未发现过。因此,PS1 基因突变通过与 GlcT-1 蛋白的异常分子相互作用导致糖脂异常。