Construction Of Sh-Ep1-Î±4Î²2-Happ695 Cell Line And Effects Of Nicotinic Agonists On Î²-Amyloid In The Cells
(1) Nicotinic acetylcholine receptors in central nervous system are thought to be new targets for Alzheimer's disease. However, the most involved nicotinic receptor subtype in Alzheimer's disease is unclear. Î±4Î²2 receptor is the most widely spread subtype in brain, involving in several important aspects of cognitive and other functions. We constructed cell line by transfecting human amyloid precursor protein (695) gene into SH-EP1 cells which have been transfected with human nicotinic receptor Î±4 subunit and Î²2 subunit gene, to observe effects of Î±4Î²2 receptors activation on Î²-amyloid, expecting to provide a new cell line for drug screening and research purpose. (2) Liposome transfection was used to express human amyloid precursor protein (695) gene in SH-EP1-Î±4Î²2 cells. Function of the transfected Î±4Î²2 receptors was tested by patch clamp. Effects of nicotine and epibatidine (selective Î±4Î²2 nicotinic receptor agonist) on Î²-amyloid were detected by Western blot and ELISA. Effects of nicotine and epibatidine on amyloid precursor protein (695) mRNA level were measured using real-time PCR. (3) Human amyloid precursor protein (695) gene was stably expressed in SH-EP1-Î±4Î²2 cells; Nicotine (1 Î¼M) and epibatidine (0.1 Î¼M) decreased intracellular and secreted Î²-amyloid in the cells; and activation of Î±4Î²2 receptors did not affect amyloid precursor protein (695) mRNA level. (4) These results suggest that the constructed cell line, expressing both amyloid precursor protein (695) gene and human nicotinic receptor Î±4 subunit and Î²2 subunit gene, might be useful for screening specific nicotinic receptor agonists against Alzheimer's disease. Alteration of AÎ² level induced by activation of Î±4Î²2 nAChR in our study might occur at a post-translational level. Â© 2007 Springer Science+Business Media, LLC.
Cellular and Molecular Neurobiology
Digital Object Identifier (DOI)
Nie, Huizhen; Li, Zuoqing; Lukas, Ronald J.; Shen, Yinghua; Song, Li; Wang, Xin; and Yin, Ming, "Construction Of Sh-Ep1-Î±4Î²2-Happ695 Cell Line And Effects Of Nicotinic Agonists On Î²-Amyloid In The Cells" (2008). Neurobiology. 198.