Abstract

The purpose of this study was to verify whether the nanosecond pulsed electric field, not eliciting thermal effects, permanently changes the molecular processes and gene expression of Leydig TM3 cells. The cells were exposed to a moderate electric field (80 quasi-rectangular shape pulses, 60 ns pulse width, and an electric field of 14 kV/cm). The putative disturbances were recorded over 24 h. After exposure to the nanosecond pulsed electric field, a 19% increase in cell diameter, a loss of microvilli, and a 70% reduction in cell adhesion were observed. Some cells showed the nonapoptotic externalization of phosphatidylserine through the pores in the plasma membrane. The cell proportion in the subG1 phase increased by 8% at the expense of the S and G2/M phases, and the DNA was fragmented in a small proportion of the cells. The membrane mitochondrial potential and superoxide content decreased by 37% and 23%, respectively. Microarray’s transcriptome analysis demonstrated a negative transient effect on the expression of genes involved in oxidative phosphorylation, DNA repair, cell proliferation, and the overexpression of plasma membrane proteins. We conclude that nanosecond pulsed electric field affected the physiology and gene expression of TM3 cells transiently, with a noticeable heterogeneity of cellular responses.

Highlights

  • Biomedical Engineering Centre, Institute of Optoelectronics, Military University of Technology, Faculty of Electronics, Military University of Technology, 00-908 Warsaw, Poland; Present address: Ecole Militaire Polytechnique, UER-ELT, Bordj El-Bahri, Algiers 16111, Algeria

  • This study aimed to evaluate the utility of nanosecond pulsed electric field

  • Our results point to a rapid sealing of the pores, indicating the transient effect of nanosecond pulsed electric field (nsPEF) on TM3 membrane permeability

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Summary

Introduction

The purpose of this study was to verify whether the nanosecond pulsed electric field, not eliciting thermal effects, permanently changes the molecular processes and gene expression of Leydig. After exposure to the nanosecond pulsed electric field, a 19% increase in cell diameter, a loss of microvilli, and a 70% reduction in cell adhesion were observed. Microarray’s transcriptome analysis demonstrated a negative transient effect on the expression of genes involved in oxidative phosphorylation, DNA repair, cell proliferation, and the overexpression of plasma membrane proteins. We conclude that nanosecond pulsed electric field affected the physiology and gene expression of TM3 cells transiently, with a noticeable heterogeneity of cellular responses. The relatively smaller pore size in the plasma membrane, due to nsPEF exposure, might result in low efficiency in the delivery of large DNA molecules (i.e., plasmids) into cells [5]. A nanosecond pulsed electric field may be more suitable for published maps and institutional affiliations

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