title: Permeabilization of the nuclear envelope following nanosecond pulsed electric field exposure.
abstract: 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.
authors.affiliation: Oak Ridge Institute for Science & Education, Joint Base San Antonio Fort Sam Houston, TX, 78234, USA. Electronic address: gary.l.thompson.3@gmail.com.
authors.forename: Gary L
authors.initials: GL
authors.lastname: Thompson
authors.affiliation: Department of Radiological Sciences, University of Texas Health Science Center at San Antonio, TX, 78234, USA.
authors.forename: Caleb C
authors.initials: CC
authors.lastname: Roth
authors.affiliation: Radio Frequency Radiation Branch, Bioeffects Division, Human Effectiveness Directorate, 711th Human Performance Wing, Air Force Research Laboratory, Joint Base San Antonio Fort Sam Houston, TX, 78234, USA.
authors.forename: Marjorie A
authors.initials: MA
authors.lastname: Kuipers
authors.affiliation: General Dynamics IT, Joint Base San Antonio Fort Sam Houston, TX, 78234, USA.
authors.forename: Gleb P
authors.initials: GP
authors.lastname: Tolstykh
authors.affiliation: Optical Radiation Branch, Bioeffects Division, Human Effectiveness Directorate, 711th Human Performance Wing, Air Force Research Laboratory, Joint Base San Antonio Fort Sam Houston, TX, 78234, USA.
authors.forename: Hope T
authors.initials: HT
authors.lastname: Beier
authors.affiliation: Radio Frequency Radiation Branch, Bioeffects Division, Human Effectiveness Directorate, 711th Human Performance Wing, Air Force Research Laboratory, Joint Base San Antonio Fort Sam Houston, TX, 78234, USA.
authors.forename: Bennett L
authors.initials: BL
authors.lastname: Ibey
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology / Q000528:radiation effects*; D016466:CHO Cells; D002463:Cell Membrane Permeability / Q000502:physiology* / Q000528:radiation effects*; D002470:Cell Survival / Q000502:physiology / Q000528:radiation effects; D006224:Cricetinae; D003412:Cricetulus; D004307:Dose-Response Relationship, Radiation; D004574:Electromagnetic Fields; D018274:Electroporation / Q000379:methods*; D009685:Nuclear Envelope / Q000502:physiology* / Q000528:radiation effects*; D011829:Radiation Dosage
keywords: Chromatin; Electroporation; H2B; PCNA; nsPEF