Fluorescence Microscope Systems to Investigate the α-synuclein Oligomerization Behavior in Living Cells

• Main Authors: Wen-Ling Chen, 陳雯齡
• Other Authors: Hsiu-Fang Fan
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/mvb6su

碩士 === 國立陽明大學 === 生命科學系暨基因體科學研究所 === 107 === Alpha-synuclein (α-Syn) is an abundant neuronal protein in the central nervous system. Moreover, it is an important factor linked to many neurodegenerative disease, including Parkinson’s disease (PD) and Lewy body dementia (LBD). α-Syn has been believed to be an unfolded monomer for a long time. However recent studies suggested that it can occur in α-helix-rich tetramers in living cells. The mutant α-Syn (A30P, E46K, H50Q, G51D, A53T) and α-Syn N103 fragment (N103) that found in the brain of PD patients can accelerate the α-Syn aggregation in vitro and in vivo. Here, we would like to develop fluorescence microscope systems to investigate the oligomerization state of interested proteins in living cells. α-Syn, mutant α-Syn and N103 fused with EGFP were transfected into HEK293T cells. The mobilities of interested protein were acquired in confocal system and analyzed with fluorescence correlation spectroscopy (FCS) to obtain their diffusion coefficients. Our preliminary data indicated that oligomerization of α-Syn is time-dependent and concentration-dependent process in HEK293T cells. Moreover, the mobility of N103 is slower than that of α-Syn and mutant α-Syn in HEK293T cells.