Association between calbindin-D28K and oogenesis in ovaries of chicken embryos in vitro.

The left ovary of chicken embryos was removed and incubated in culture medium with a thymidine analogue, bromodeoxyuridine (BrdU), in vitro. In addition, fertile chicken eggs were injected with BrdU via the extraembryonic vessels and incubated for 24 hours. The ovaries were then processed for immunohistochemical localization of calbindin-D28K (a 28-kd vitamin D-dependent calcium-binding protein) and BrdU. Calbindin-D28K was detected in the germinal epithelium and in cells surrounding the oogonia and oocytes (future granulosa cells) of the embryonic chicken ovary. However, BrdU was observed in the nucleus of the oogonia and oocytes of the chicken embryonic ovaries. Comparison of the 2 adjacent sections, immunostained for calbindin-D28K and BrdU consecutively, indicated that BrdU, the marker for cell proliferation was not detected in calbindin-D28K-containing cells, namely, germinal epithelium and future granulosa cells, in the ovary of chicken embryos. These results suggested that calbindin-D28K-containing cells in the ovary were not in the process of cell division during the 24-hour incubation of chicken embryos.