Abstract
To study the ultrastructure of nuclear pore complexes (NPCs), a wide spectrum of different electron microscopy (EM) or atomic force microscopy (AFM) techniques can be employed. The combination of these methods can reveal new insights into the structural and functional organization of this important supramolecular machine through which nucleocytoplasmic transport occurs. Negative staining, quick freezing/freeze-drying/rotary metal shadowing, embedding and thin sectioning, cryoelectron microscopy and tomography, scanning electron microscopy, or combination with immunolabeling techniques are tools for collecting data and information about the three-dimensional structure and architecture of the NPCs. AFM enables investigation of the functional dynamics of native NPCs under physiological conditions.
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References
Unwin, P. N. and Milligan, R. A. (1982) A large particle associated with the perimeter of the nuclear pore complex. J. Cell Biol. 93, 63–75.
Hinshaw, J. E., Carragher, B. O., and Milligan, R. A. (1992) Architecture and design of the nuclear pore complex. Cell 69, 1133–1141.
Akey, C. W. and Radermacher, M. (1993) Architecture of the Xenopus nuclear pore complex revealed by 3-dimensional cryo-electron microscopy. J. Cell Biol. 122, 1–19.
Stoffler, D., Feja, B., Fahrenkrog, B., Walz, J., Typke, D., and Aebi, U. (2003) Cryo-electron tomography provides novel insights into nuclear pore architecture: implications for nucleocytoplasmic transport. J. Mol. Biol. 328, 119–130.
Reichelt, R., Holzenburg, A., Buhle, E. L., Jr., Jarnik, M., Engel, A., and Aebi, U. (1990) Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components. J. Cell Biol. 110, 883–894.
Fahrenkrog, B., Köser, J., and Aebi, U. (2004) The nuclear pore complex: a jack of all trades? Trends Cell Biol 29, 175–182.
Fahrenkrog, B. and Aebi, U. (2003) The nuclear pore complex: nucleocytoplasmic transport and beyond. Nat. Rev. Mol. Cell Biol. 4, 757–766.
Jarnik, M. and Aebi, U. (1991) Toward a more complete 3-D structure of the nuclear pore complex. J. Struct. Biol. 107, 291–308.
Hinshaw, J. E. and Milligan, R. A. (2003) Nuclear pore complexes exceeding eightfold rotational symmetry. J. Struct. Biol. 141, 259–268.
Harris, J. R., ed. (1997) Negative Staining and Cryoelectron Microscopy: The Thin Film Techniques, BIOS Scientific Publishers Limited, Oxford, UK.
Goldberg, M. W. and Allen, T. D. (1992) High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores. J. Cell Biol. 119, 1429–1440.
Goldberg, M. W. and Allen, T. D. (1993) The nuclear pore complex: three-dimensional surface structure revealed by field emission, in-lens scanning electron microscopy, with underlying structure uncovered by proteolysis. J. Cell Sci. 106, 261–274.
Goldberg, M. W., Wiese, C, Allen, T. D., and Wilson, K. L. (1997) Dimples, pores, starrings, and thin rings on growing nuclear envelopes: evidence for structural intermediates in nuclear pore complex assembly. J. Cell Sci. 110, 409–420.
Panté, N., Bastos, R., McMorrow, I., Burke, B., and Aebi, U. (1994) Interactions and three-dimensional localization of a group of nuclear pore complex proteins. J. Cell Biol. 126,603–617.
Fahrenkrog, B., Maco, B., Fager, A. M., Köser, J., Sauder, U., Ullman, K.S., and Aebi, U. (2002) Domain-specific antibodies reveal multiple-site topology of Nup153 within the nuclear pore complex. J. Struct. Biol. 140, 254–267.
Slot, J. W. and Geuze, H. J. (1985) A new method of preparing gold probes for multiple-labeling cytochemistry. Eur. J. Cell Biol. 38, 87–93.
Baschong, W. and Wrigley, N. G. (1990) Small colloidal gold conjugated to Fab fragments or to immunoglobulin G as high-resolution labels for electron microscopy: a technical overview. J. Electron Microsc. Tech. 14, 313–323.
Stoffler, D., Goldie, K. N., Feja, B., and Aebi, U. (1999) Calcium-mediated structural changes of native nuclear pore complexes monitored by time-lapse atomic force microscopy. J. Mol. Biol. 287, 741–752.
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© 2006 Humana Press Inc., Totowa, NJ
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Maco, B., Fahrenkrog, B., Huang, NP., Aebi, U. (2006). Nuclear Pore Complex Structure and Plasticity Revealed by Electron and Atomic Force Microscopy. In: Liu, X.J. (eds) Xenopus Protocols. Methods in Molecular Biology™, vol 322. Humana Press. https://doi.org/10.1007/978-1-59745-000-3_19
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DOI: https://doi.org/10.1007/978-1-59745-000-3_19
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