Nuclear envelope barrier leak induced by dexamethasone.
Nuclear pore complexes (NPCs) are multiprotein channels that span the nuclear envelope. They strongly limit the efficiency of gene transfection by restriction of nuclear delivery of exogenously applied therapeutic macromolecules. NPC dilation could significantly increase this efficiency. Recently, it was shown in oocytes of Xenopus laevis that NPCs dilate from about 82 to 110 nm within min after injection of the glucocorticoid analog dexamethasone (dex). In the present paper we analyzed by means of atomic force microscopy the structural details of NPC dilation and correlated them with functional changes in nuclear envelope permeability. 5-11 min after Dex injection NPC dilation was found at its maximum (approximately 140 nm). In addition, a yet unknown configuration, so-called giant pore, up to 300 nm in diameter, was visualized. Giant pore formation was paralleled by an increase in nuclear envelope permeability tested by electrophysiology and confocal fluorescence microscopy. Even large macromolecules lacking any nuclear localization signal (77 kDa FITC-dextran, molecule diameter up to 36 nm) could gain access to the nucleus. We conclude that dex transiently opens unspecific pathways for large macromolecules. Dex treatment could be potentially useful for improving the efficiency of nuclear gene transfection.
Kastrup
Lilian
L
Institute of Physiology II, University of Muenster, Muenster, Germany.
Oberleithner
Hans
H
Ludwig
Yvonne
Y
Schafer
Claudia
C
Shahin
Victor
V
D000818:Animals; D003907:Dexamethasone / Q000494:pharmacology*; D003911:Dextrans / Q000493:pharmacokinetics* / Q000494:pharmacology; D005260:Female; D018625:Microscopy, Atomic Force; D009685:Nuclear Envelope / Q000737:chemistry* / Q000378:metabolism*; D022022:Nuclear Pore / Q000187:drug effects*; D009865:Oocytes / Q000187:drug effects; D010539:Permeability; D014981:Xenopus
D003911:Dextrans; D003907:Dexamethasone