Abstract
Electron microscopy allows direct visualization of the underlying organization of cell surface components on a nano-scale. Immuno-gold labelling of isolated plasma membranes generates point patterns that enable mapping of protein and lipid distributions. 2D spatial statistics reveals the extent to which these distributions are clustered or dispersed and allows the extent of co-localization between different cell surface components to be precisely determined. This approach has been successfully applied to the study of signalling network organization and the consequences of physiological changes in modulating cell surface function.
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References
Kusumi A, Fujiwara TK, Chadda R, Xie M, Tsunoyama TA et al (2012) Dynamic organizing principles of the plasma membrane that regulate signal transduction: commemorating the fortieth anniversary of Singer and Nicolson's fluid-mosaic model. Annu Rev Cell Dev Biol 28:215–250
Parton RG, del Pozo MA (2013) Caveolae as plasma membrane sensors, protectors and organizers. Nat Rev Mol Cell Biol 14:98–112
Soboloff J, Rothberg BS, Madesh M, Gill DL (2012) STIM proteins: dynamic calcium signal transducers. Nat Rev Mol Cell Biol 13:549–565
Insall RH, Machesky LM (2009) Actin dynamics at the leading edge: from simple machinery to complex networks. Dev Cell 17:310–322
Laude AJ, Prior IA (2004) Plasma membrane microdomains: organization, function and trafficking. Mol Membr Biol 21:193–205
Simons K, Sampaio JL (2011) Membrane organization and lipid rafts. Cold Spring Harb Perspect Biol 3:a004697
Munro S (2003) Lipid rafts: elusive or illusive? Cell 115:377–388
Prior IA, Muncke C, Parton RG, Hancock JF (2003) Direct visualization of Ras proteins in spatially distinct cell surface microdomains. J Cell Biol 160:165–170
Prior IA, Parton RG, Hancock JF (2003) Observing cell surface signaling domains using electron microscopy. Sci STKE 2003:PL9
Zhang J, Leiderman K, Pfeiffer JR, Wilson BS, Oliver JM et al (2006) Characterizing the topography of membrane receptors and signaling molecules from spatial patterns obtained using nanometer-scale electron-dense probes and electron microscopy. Micron 37:14–34
Wilson BS, Steinberg SL, Liederman K, Pfeiffer JR, Surviladze Z et al (2004) Markers for detergent-resistant lipid rafts occupy distinct and dynamic domains in native membranes. Mol Biol Cell 15:2580–2592
Cho KJ, Kasai RS, Park JH, Chigurupati S, Heidorn SJ et al (2012) Raf inhibitors target ras spatiotemporal dynamics. Curr Biol 22:945–955
Chapkin RS, Wang N, Fan YY, Lupton JR, Prior IA (2008) Docosahexaenoic acid alters the size and distribution of cell surface microdomains. Biochim Biophys Acta 1778: 466–471
Slot JW, Geuze HJ (1985) A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol 38:87–93
Besag JE (1977) Contribution to the discussion of Dr Ripley’s paper. J R Stat Soc B39:193–195
Diggle PJ (1986) Displaced amacrine cells in the retina of a rabbit: analysis of a bivariate spatial point pattern. J Neurosci Methods 18:115–125
Ripley BD (1979) Tests of randomness for spatial point patterns J. J R Stat Soc B41: 368–374
Ripley RD (1977) Modelling spatial patterns. J R Stat Soc B39:172–192
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Beckett, A.J., Prior, I.A. (2015). Electron Microscopy Methods for Studying Plasma Membranes. In: Owen, D. (eds) Methods in Membrane Lipids. Methods in Molecular Biology, vol 1232. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1752-5_12
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DOI: https://doi.org/10.1007/978-1-4939-1752-5_12
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