Nucleologenesis in the cleaving bovine embryo: immunocytochemical aspects

In vivo nucleologenesis was studied in bovine embryos by electron microscopic immunogold labelling of DNA, RNA, protein C23 and protein B23. We have used the classification of Kopecný et al. (1989b) and Kopecný (1990) dividing nucleologenesis in four steps: compact nucleolar precursor body (NPB), monovacuolated NPB, NPB containing secondary vacuoles and fully reticulated nucleolus. These different features of early bovine embryo nucleologenesis were mainly observed during the eight-cell stage. In the first step of nucleolar development, the association of compact NPB with DNA structures was observed. DNA was also labelled in some small secondary vacuoles appearing during the third developmental step. From the second step onward, the labelling of protein C23 was observed in the compact fibrillar network of the NPB. Protein B23 started to be labelled in the compact fibrillar mass at the third step. RNA labelling was also observed for the first time in NPB containing secondary vacuoles. Labelled RNA was located in the peripheral region of compact fibrillar mass as well as along the border of the vacuoles. In the reticulated nucleolus, the dense fibrillar component was found to contain both proteins and RNA.