Abstract
Application of nanosecond pulsed electric fields (nsPEFs) has attracted rising attention in various scientific fields including medical, pharmacological, and biological sciences, although its effects and molecular mechanisms leading to the effects remain poorly understood. Here, we show that a single, high-intensity (10–30 kV/cm), 60-ns PEF exposure affects gene expression and impairs development of eyes and germ cells in medaka (Oryzias latipes). Exposure of early blastula stage embryos to nsPEF down-regulated the expression of several transcription factors which are essential for eye development, causing abnormal eye formation. Moreover, the majority of the exposed genetic female embryos showed a fewer number of germ cells similar to that of the control (unexposed) genetic male at 9 days post-fertilization (dpf). However, all-trans retinoic acid (atRA) treatment following the exposure rescued proliferation of germ cells and resumption of normal eye development, suggesting that the phenotypes induced by nsPEF are caused by a decrease of retinoic acid levels. These results confirm that nsPEFs induce novel effects during embryogenesis in medaka.
Highlights
Electroporation is routinely used in the field of molecular biology for the transfer of DNA to cell and tissues [1,2]
Our results show that the nanosecond pulsed electric fields (nsPEFs) affects the development of eyes and germ cells in medaka
The exposed embryos showed several defective eye phenotypes such as small, single, or the complete absence of eyes during early embryogenesis (Figs. 1C–G and Figs. 4D–F). pax6 gene is known as a key regulator of vertebrate eye development [24], while in medaka rx2 gene’s role in eye development has not yet been determined
Summary
Electroporation is routinely used in the field of molecular biology for the transfer of DNA to cell and tissues [1,2]. Effects of nsPEFs on cells differ from millisecond or microsecond duration electric fields. Recent discoveries have suggested new possibilities of nsPEFs use for cancer treatment [9,10], while nsPEF application to cells and tissue has demonstrated potential to induce further unique modifications [11,12]. This background illustrates the requirement for insight on underlying molecular mechanisms
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