Permeabilization of the nuclear envelope following nanosecond pulsed electric field exposure

• Published in: Biochemical and biophysical research communications 2016-01, Vol.470 (1), p.35-40
• Main Authors: Thompson, Gary L., Roth, Caleb C., Kuipers, Marjorie A., Tolstykh, Gleb P., Beier, Hope T., Ibey, Bennett L.
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; ABSORBED RADIATION DOSES; Animals; ANTIGENS; APOPTOSIS; Apoptosis - physiology; Apoptosis - radiation effects; CELL CULTURES; Cell Membrane Permeability - physiology; Cell Membrane Permeability - radiation effects; CELL MEMBRANES; CELL NUCLEI; Cell Survival - physiology; Cell Survival - radiation effects; CHINA; CHO CELLS; CHROMATIN; Cricetinae; Cricetulus; DNA; Dose-Response Relationship, Radiation; Electromagnetic Fields; Electroporation; Electroporation - methods; FLUORESCENCE; H2B; HAMSTERS; HISTONES; nsPEF; NUCLEAR DAMAGE; Nuclear Envelope - physiology; Nuclear Envelope - radiation effects; OVARIES; PCNA; Radiation Dosage
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2015.12.092

Permeabilization of cell membranes occurs upon exposure to a threshold absorbed dose (AD) of nanosecond pulsed electric fields (nsPEF). The ultimate, physiological bioeffect of this exposure depends on the type of cultured cell and environment, indicating that cell-specific pathways and structures are stimulated. Here we investigate 10 and 600 ns duration PEF effects on Chinese hamster ovary (CHO) cell nuclei, where our hypothesis is that pulse disruption of the nuclear envelope membrane leads to observed cell death and decreased viability 24 h post-exposure. To observe short-term responses to nsPEF exposure, CHO cells have been stably transfected with two fluorescently-labeled proteins known to be sequestered for cellular chromosomal function within the nucleus – histone-2b (H2B) and proliferating cell nuclear antigen (PCNA). H2B remains associated with chromatin after nsPEF exposure, whereas PCNA leaks out of nuclei permeabilized by a threshold AD of 10 and 600 ns PEF. A downturn in 24 h viability, measured by MTT assay, is observed at the number of pulses required to induce permeabilization of the nucleus. •The ability of nsPEF to damage nuclear structures within cells is investigated.•Leakage of proliferating nuclear antigen from nuclei is induced by nsPEF.•High doses of nsPEF disrupt cortical lamin and cause chromatin decompaction.•Histone H2B remains attached to chromatin following nsPEF exposure.•DNA does not leak out of nsPEF-permeabilized nuclei.