Apoptosis is associated with formation and persistence of the embryonic fissure

The role of apoptosis in the transitory ocular embryonic structures has not been clarified yet, therefore, in the present study we focused on one of the transitory ocular structures, the embryonic fissure. To elucidate the developmental mechanism of the embryonic fissure, we observed cell death by apoptosis in the optic cup during early development in mice.

Pairs of C57BL6N/Jcl mice, each comprising an estrous female and a potent male, were caged together overnight. Females that had vaginal plugs the next morning were considered at day 0 of pregnancy. The embryos or fetuses were removed by laparotomy on days 9, 10, 11, 12, 13, 14, 16, and 18 of gestation. Tissue blocks of the eyes were fixed and embedded in paraffin wax. Serial frontal sections of the eye were cut and stained with the TUNEL method and then counterstained by hematoxylin or methyl green solution. We examined TUNEL-positive cells in the optic cup by light microscopy.

TUNEL-positive cells were seen at the lower nasal side of the optic cup, corresponding to the presumed embryonic fissure area, on day 9 of gestation before the formation of the embryonic fissure. Many TUNEL-positive cells were present at the lips of the embryonic fissure on days 10, 11, and 12. In contrast, TUNEL-positive cells were not detectable in the corresponding area on day 13 after the complete closure of the embryonic fissure.

Apoptosis is anatomically closely associated, and appears to be essential for the formation and persistence of the embryonic fissure.