Nuclear transport by laser-induced pressure transients.
OBJECTIVE
Control of the transport of molecules into the nucleus represents a key regulatory mechanism for differentiation, transformation, and signal transduction. Permeabilization of the nuclear envelope by physical methods can have applications in gene therapy. Laser-induced pressure transients can produce temporary aqueous pores analogous to those produced by electroporation and that the cells can survive this procedure. In this study, we examine the role of the pressure transients in creating similar pores in the nuclear envelope.


METHODS
The target human peripheral blood mononuclear cells in a 62 microM 72 kDa fluoresceinated dextran solution were exposed to the pressure transients generated by laser ablation. An in vitro fluorescence confocal microscope was used to visualize and quantify the fluoresceinated dextran in the cytoplasmic and nuclear compartments.


RESULTS
In contrast to electroporation, the pressure transients could deliver 72 kDa fluoresceinated dextrans, which are normally excluded by the nucleus, across the nuclear envelope into the nucleus. In addition to creating pores in the plasma membrane, temporary pores were also created in the nuclear envelope following exposure to pressure transients.


CONCLUSIONS
The production of temporary nuclear pores could provide a unique resource for drug-delivery and gene therapy.
Lin
Tai-Yuan David
TY
Wellman Laboratories of Photomedicine, Department of Photopathology and Dermatology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
McAuliffe
Daniel J
DJ
Michaud
Norm
N
Zhang
Hong
H
Lee
Shun
S
Doukas
Apostolos G
AG
Flotte
Thomas J
TJ
D002467:Cell Nucleus / Q000378:metabolism* / Q000528:radiation effects; D004396:Coloring Agents; D003593:Cytoplasm / Q000528:radiation effects / Q000648:ultrastructure; D004058:Diffusion; D018274:Electroporation; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D018613:Microscopy, Confocal; D009685:Nuclear Envelope / Q000378:metabolism / Q000528:radiation effects / Q000648:ultrastructure; D010539:Permeability / Q000528:radiation effects; D011312:Pressure; D011419:Propidium; D014162:Transfection
D004396:Coloring Agents; D011419:Propidium