The nuclear pore complex: three-dimensional surface structure revealed by field emission, in-lens scanning electron microscopy, with underlying structure uncovered by proteolysis.
AffiliationCRC Department of Structural Cell Biology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.
MetadataShow full item record
AbstractThe structure of the nuclear pore complex (NPC) has been previously studied by many different electron microscopic techniques. Recently, scanning electron microscopes have been developed that can visualise biologically relevant structural detail at the same level of resolution as transmission electron microscopes and have been used to study NPC structure. We have used such an instrument to visualise directly the structure of both cytoplasmic and nucleoplasmic surfaces of the NPC of manually isolated amphibian oocyte nuclear envelopes that have been spread, fixed, critical point dried and coated with a thin fine-grained film of chromium or tantalum. We present images that directly show features of the NPC that are visible at each surface, including coaxial rings, cytoplasmic particles, plug/spoke complexes and the nucleoplasmic basket or fishtrap. Some cytoplasmic particles are rod-shaped or possibly "T"-shaped, can be quite long structures extending into the cytoplasm and may be joined to the coaxial ring at a position between each subunit. Both coaxial rings, which are proud of the membranes, can be exposed by light proteolytic digestion, revealing eight equal subunits each of which may be bipartite. We have determined that the nucleoplasmic filaments that make up the baskets are attached to the outer periphery of the coaxial ring at a position between each of its subunits. These filaments extend into the nucleoplasm and insert at the distal end to the smaller basket ring. The space left between adjacent basket filaments would exclude particles bigger than about 25 nm, which is consistent with the exclusion limit previously found for NPC-transported molecules.
CitationThe nuclear pore complex: three-dimensional surface structure revealed by field emission, in-lens scanning electron microscopy, with underlying structure uncovered by proteolysis. 1993, 106 ( Pt 1):261-74 J. Cell. Sci.
JournalJournal of Cell Science
- The nuclear pore complex and lamina: three-dimensional structures and interactions determined by field emission in-lens scanning electron microscopy.
- Authors: Goldberg MW, Allen TD
- Issue date: 1996 Apr 12
- 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.
- Authors: Goldberg MW, Allen TD
- Issue date: 1992 Dec
- Nuclear pore complex structure in birds.
- Authors: Goldberg MW, Solovei I, Allen TD
- Issue date: 1997 Aug
- Active nuclear pore complexes in Chironomus: visualization of transporter configurations related to mRNP export.
- Authors: Kiseleva E, Goldberg MW, Allen TD, Akey CW
- Issue date: 1998 Jan
- Accessing nuclear structure for field emission, in lens, scanning electron microscopy (FEISEM).
- Authors: Allen TD, Bennion GR, Rutherford SA, Reipert S, Ramalho A, Kiseleva E, Goldberg MW
- Issue date: 1996