Insights into Cell Membrane Microdomain Organization from Live Cell Single Particle Tracking of the IgE High Affinity Receptor FcϵRI of Mast Cells
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Current models propose that the plasma membrane of animal cells is composed of heterogeneous and dynamic microdomains known variously as cytoskeletal corrals, lipid rafts and protein islands. Much of the experimental evidence for these membrane compartments is indirect. Recently, live cell single particle tracking studies using quantum dot-labeled IgE bound to its high affinity receptor FcϵRI, provided direct evidence for the confinement of receptors within micrometer-scale cytoskeletal corrals.
In this study, we show that an innovative time-series analysis of single particle tracking data for the high affinity IgE receptor, FcϵRI, on mast cells provides substantial quantitative information about the submicrometer organization of the membrane. The analysis focuses on the probability distribution function of the lengths of the jumps in the positions of the quantum dots labeling individual IgE FcϵRI complexes between frames in movies of their motion. Our results demonstrate the presence, within the micrometer-scale cytoskeletal corrals, of smaller subdomains that provide an additional level of receptor confinement. There is no characteristic size for these subdomains; their size varies smoothly from a few tens of nanometers to a over a hundred nanometers.
In QD-IGE labeled unstimulated cells, jumps of less than 70 nm predominate over longer jumps. Addition of multivalent antigen to crosslink the QD-IgE-FcϵRI complexes causes a rapid slowing of receptor motion followed by a long tail of mostly jumps less than 70 nm. The reduced receptor mobility likely reflects both the membrane heterogeneity revealed by the confined motion of the monomeric receptor complexes and the antigen-induced cross linking of these complexes into dimers and higher oligomers. In both cases, the probability distribution of the jump lengths is well fit, from 10 nm to over 100 nm, by a novel power law. The fit for short jumps suggests that the motion of the quantum dots can be modeled as diffusion in a fractal space of dimension less than two.
KeywordsLive cell IgE-FcϵRI Microdomains Cytoskeletal corrals Single particle tracking Quantum dots Time series Jump sizes Standard deviation of jumps Time-dependent diffusion coefficient
- Andrews, N. L. (2011). The role of diffusion and membrane topography in the initiation of high affinity IgE receptor signaling. PhD thesis, University of New Mexico, Albuquerque, New Mexico, USA. Google Scholar
- Bachir, A. L. (2006). Characterization of blinking dynamics in quantum dots ensembles using image correlation spectroscopy. J. Appl. Phys., 99(6). doi:10.1063/1.2175470.
- Kubitscheck, U. (2009). Single molecule fluorescence monitoring in eukaryotic cells: intranuclear dynamics of splicing factors (pp. 1–17). Wiley-VCH Verlag GmbH & Co. KGaA. Google Scholar
- Kusumi, A., Nakada, C., Ritchie, K., Murase, K., Suzuki, K., Murakoshi, H., Kasai, R. S., Kondo, J., & Fujiwara, T. (2005). Paradigm shift of the plasma membrane concept from the two-dimensional continuum fluid to the partitioned fluid: high-speed single-molecule tracking of membrane molecules. Annu. Rev. Biophys. Biomol. Struct., 34(1), 351–378. CrossRefGoogle Scholar
- Lidke, D. S., Andrews, N. L., Pfeiffer, J. R., Jones, H. D. T., Sinclair, M. B., Haaland, D. M., Burns, A. R., Wilson, B. S., Oliver, J. M., & Lidke, K. A. (2007). Exploring membrane protein dynamics by multicolor single quantum dot imaging using wide field, TIRF, and hyperspectral microscopy. Proc. SPIE, 6448, 6448. Google Scholar
- Pike, L. J. (2006). A report on the keystone symposium on lipid rafts and cell function. PubMed, 47(7), 1597–1598. Google Scholar
- Wilson, B. S., Pfeiffer, J. R., & Oliver, J. M. (2002). FcϵRI signaling observed from the inside of the mast cell membrane. Mol. Immunol., 1144, 1–10. Google Scholar