Analysis of 伪3 GlyR single particle tracking in the cell membrane

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• 作者：Kristof Notelaers ; Susana Rocha ; Rik Paesen ; Nick Smisdom ; Ben De Clercq ; Jochen C. Meier ; Jean-Michel Rigo ; Johan Hofkens ; Marcel Ameloot
• 关键词：SPT ; Single particle tracking ; GlyR ; Glycine receptor ; LGIC ; Ligand gated ion channel ; TM ; Transmembrane segment ; HEK 293 ; Human embryonic kidney 293 ; MSD ; Mean square displacement ; TIRF ; Total internal reflection fluorescence
• 刊名：Biochimica et Biophysica Acta (BBA)/Molecular Cell Research
• 出版年：March, 2014
• 年：2014
• 卷：1843
• 期：3
• 页码：544-553
• 全文大小：927 K

文摘

Single particle tracking (SPT) of transmembrane receptors in the plasma membrane often reveals heterogeneous diffusion. A thorough interpretation of the displacements requires an extensive analysis suited for discrimination of different motion types present in the data. Here the diffusion pattern of the homomeric 伪3-containing glycine receptor (GlyR) is analyzed in the membrane of HEK 293 cells. More specifically, the influence of the 伪3 RNA splice variants 伪3K and 伪3L on lateral membrane diffusion of the receptor is revealed in detail. Using a combination of ensemble and local SPT analysis, free and anomalous diffusion parameters are determined. The GlyR 伪3 free diffusion coefficient is found to be 0.13 卤 0.01 渭m²/s and both receptor variants display confined motion. The confinement probability level and residence time are significantly elevated for the 伪3L variant compared to the 伪3K variant. Furthermore, for the 伪3L GlyR, the presence of directed motion was also established, with a velocity matching that of saltatory vesicular transport. These findings reveal that 伪3 GlyRs are prone to different types of anomalous diffusion and reinforce the role of RNA splicing in determining lateral membrane trafficking.