Effects of nanosecond pulsed electric field on the embryonic development of medaka fish egg (Oryzias latipes)

Abstract

The paper describes the effects of 50 to 300 nanosecond pulsed electric field on the embryonic development of the medaka fish egg (Oryzias latipes). In the recent years it has been reported that applying short (less than 100 ns) pulses increased the possibility of electric field interactions with subcellular structures, which leaded to secondary cellular events, such as temporally increase in cell membrane permeability and induction of apoptosis. The goal of the current study was to find the effects of short pulsed electric field in-vivo during embryo development. A pulsed power modulator using a magnetic compression circuit was employed to generat 0.5 to 20 kV pulses with 50 to 300 nanosecond pulse durations. Input voltage and current were measured by using an oscilloscope and a current monitor. Fertilized eggs of d-rR medaka were used. The age of the experimental eggs were 6 hours, 1 day and 2 days post fertilization. In each experiment, a single medaka egg (about 1.2 mm diameter) was set at the middle of a 2 mm or 4 mm cuvette and a single electric pulse was applied. After the experiments the eggs were observed under a microscope until they hatched or died. A fluorescent plasma membrane integrity indicator, propidium iodide (PI), was used to study electroporative uptake kinetics of the embryo cells after the electric pulse exposure. By applying 300 ns electric pulses, extensive damage of eggs were observed immediately after pulse application. For shorter 50 ns width pulses and low electric field, delayed hatching consistent with electric field intracellular interaction was observed, whereas stronger electric field affected the eggs immediately after the pulse and those eggs could not survive and died a few days later.