Monitoring the permeability of the nuclear envelope during the cell cycle.
In animal organisms the nuclear envelope (NE) dis-assembles during cell division resulting in complete intermixing of cytoplasmic and nuclear compartments. This leads to the activation of many mitotic enzymes, which were kept away from their substrates or regulators by nuclear or cytoplasmic sequestration in interphase. Nuclear envelope breakdown (NEBD) is thus an essential step of mitotic entry and commits a cell to M-phase. NEBD begins with the partial disassembly of nuclear pore complexes, leading to a limited permeabilization of the NE for molecules up to approximately 40 nm diameter. This is followed by the complete disruption of nuclear pores, which causes local fenestration of the double nuclear membrane and subsequently breakdown of the entire NE structure. Here, we describe the use of different sized inert fluorescent tracer molecules to directly visualize these different steps of NEBD in live cells by fluorescence microscopy.
Lénárt
Péter
P
Gene Expression and Cell Biology/Biophysics Programmes, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Ellenberg
Jan
J
D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D003911:Dextrans / Q000737:chemistry; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D009685:Nuclear Envelope / Q000378:metabolism*; D010539:Permeability; D045663:Quantum Dots; D011993:Recombinant Fusion Proteins / Q000032:analysis
D003911:Dextrans; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins