In vitro measurement of nuclear permeability changes in apoptosis

In the eukaryotic cell, exchange of biomolecules between nucleus and cytoplasm is a highly regulated process which responds sensitively to changes of the environment. One well-known cellular response to environmental challenges is cell death by apoptosis. In fact, apoptosis has been shown to affect...

Full description

Saved in:
Bibliographic Details
Published in:Analytical biochemistry 2003-07, Vol.318 (2), p.244-253
Main Authors: Roehrig, Sebastian, Tabbert, Anja, Ferrando-May, Elisa
Format: Article
Language:English
Subjects:
Active Transport, Cell Nucleus
Apoptosis
Confocal microscopy
Dextran
Digitonin
Fluorescence imaging
HeLa Cells
Humans
Jurkat Cells
Microscopy, Confocal
Nuclear Envelope - metabolism
Nuclear pore
Nuclear transport
Permeability
Software
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In the eukaryotic cell, exchange of biomolecules between nucleus and cytoplasm is a highly regulated process which responds sensitively to changes of the environment. One well-known cellular response to environmental challenges is cell death by apoptosis. In fact, apoptosis has been shown to affect the nucleocytoplasmic transport machinery, in particular the nuclear pore, by modulating its size exclusion limit for passive diffusion. The underlying molecular factors are still unknown, mainly because of the lack of a suitable system to detect and quantitate the apoptotic effects on the nuclear pore. Here we present an assay that was designed to measure alterations of the permeability of the nuclear envelope under apoptotic conditions. The assay is based on the well-established technique of selective permeabilization of the plasma membrane with digitonin and allows assessment of permeability changes in nonfixed samples. It comprises a computer program, called Nuclear Permeability Assay, for the quantitation of the nuclear fluorescence signal, which may be generally employed for the evaluation of in vitro transport systems using semipermeabilized cells, such as assays for nuclear import and export.
ISSN:0003-2697
1096-0309
DOI:10.1016/S0003-2697(03)00242-2