Effects of nsPEF on development of germ cells.

<p>(A) Photographs show DsRed- or GFP-positive germ cells in the individual medaka embryos at 9 dpf; control, 30 kV/cm nsPEF-exposed, and atRA-treated following 30 kV/cm exposure. Arrows and arrowheads indicate GFP-positive germ cells and autofluorescence of XY-specific leucophores that were ascertained under a different excitation wavelength (GFP2), respectively. (B) The number of DsRed-positive germ cells in control and nsPEF-exposed medaka at 9 dpf and (C) the number of DsRed-positive germ cells when the egg was placed in a different orientation between the two electrodes and exposed to 30 kV/cm nsPEF. L, R and U indicate that egg blastomeres directed towards the left electrode (the high voltage electrode), the right electrode (ground) and upward, respectively. (<i>n</i> ≥ 5; *<i>P</i><0.05, **<i>P</i><0.01). Black circles indicate embryos with GFP-positive germ cells and white circles refer to those without GFP-positive germ cells. (D-F) Embryos at 4 dpf; D: the left and right side photos show a control and a 30 kV/cm nsPEF exposed embryos, respectively, E: a 30 kV/cm nsPEF exposed embryo, and F: an embryo treated with atRA after 30 kV/cm exposure. Scale bar 100 µm.