Glycogen synthase kinase-3 beta overexpression enhances tau protein phosphorylation and attenuates tubulin acetylation in SH-SY5Y cells
- VernacularTitle:SH-SY5Y细胞高表达糖原合成激酶3?对tau蛋白磷酸化和微管稳定性的影响
- Author:
Jie SHEN
;
Xiaochun CHEN
;
Xu LIN
;
Jing ZHANG
- Publication Type:Journal Article
- Keywords:
Glycogen synthase kinase-3?;
tau proteins;
Phosphorylation;
Tubulin
- From:
Chinese Journal of Pathophysiology
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To construct GSK3?-overexpressed SH-SY5Y cells and to observe the effects of GSK3?-overexpression on tau protein phosphorylation and tubulin acetylation in SH-SY5Y engineered cells. METHODS: The cDNA of GSK3? construct was subcloned into mammalian expression vector pBudCE4.1. The integrity of the GSK3? construct was confirmed by sequence analysis. GSK3? was transiently transfected into SH-SY5Y cells using Lipofectamine2000. Western blotting was used to measured protein levels of GSK3? and phosphorylating GSK3?, as well as, the total tau and phosphorylated tau protein and acetylated tubulin. RESULTS: 36 h after transfection, the levels of GSK3? and phosphorylating GSK3? in SH-SY5Y cells were significantly increased compared with non-transfection group and vector group. After 48 h, the levels of phosphorylated tau protein (Ser199/202, Thr231 and Thr205 residues) but not total tau protein were markedly increased in GSK3?-overexpressed SH-SY5Y cells. In addition, the level of acetylated tubulin was lower than that in non-transfection group and vector group. CONCLUSION: The over-expression of GSK3? in SH-SY5Y cells results in robust increases in tau protein phosphorylation at Ser199/202, Thr231 and Thr205 residues, and decreases in tubulin acetylation.
