To prepare live gonadal ridges and ovaries at the time of staining (n=10), we kept them in the Dulbecco Minimum Essential Medium (DMEM, Gibco, 11995), which contained 1.5 [micro]M/mL PI at 37[degrees]C and 5% C[O.sub.2] in a humidified incubator for 30 minutes in the dark.
Morphological Features of the Embryonic Gonadal Ridges and the Neonatal Ovaries of the Mice
Somatic and germ cells dispersed throughout the gonadal ridges, and follicular arrangement was not observed (figure 1).
In the embryonic gonadal ridges, TUNEL-positive germ cells were detected (figure 3A).
Autophagic vacuoles were shown as green fluorescent dot-like structures spreading within the cytoplasm of the germ cells in the embryonic gonadal ridges (figure 4).
In the urogenital ridges of the embryos, necrotic cell death was seen in both mesonephric regions and gonadal ridges. Necrosis was observed in both germ and somatic cells (figure 6).
Percentage of Programmed Cell Death in the Embryonic Gonadal Ridges and the Neonatal Ovaries of the Mice