Fate of ciliated epidermal cells during early development of Xenopus laevis using whole-mount immunostaining with an antibody against chondroitin 6-sulfate proteoglycan and anti-tubulin: transdifferentiation or metaplasia of amphibian epidermis

Xenopus embryonic epidermis changes its cellular composition during development: the appearance of ciliated epidermal cells before hatching is a remarkable characteristic. In this study, the functional change of ciliated cells to mucus-secreting cells was examined with immunocytochemistry using anti-tubulin and anti-chondroitin 6-sulfate (C6S). Before hatching, most epidermal cells were labeled with anti-C6S in a granular fashion. Immunoelectron microscopy revealed that the anti-C6S-positive structure was the mucus granule. Ciliated epidermal cells lacked anti-C6S staining, but were strongly labeled with anti-tubulin. After hatching, most ciliated cells in the surface of the embryo disappeared. During their disappearance, some ciliated cells exhibited anti-C6S-positive granular labeling. This strongly suggests that the disappearance of ciliated cells is a functional conversion to mucus-secreting cells instead of shedding through cell death.     

Ontogeny and tissue distribution of leukocyte-common antigen bearing cells during early development of Xenopus laevis

To analyze the ontogenic emergence of leukocytes during early development, a mouse monoclonal antibody (IgG1), designated as XL-1, was produced against the peritoneal macrophages of adult Xenopus laevis. The XL-1 determinant was expressed on all types of leukocytes, including lymphocytes, granulocytes, thrombocytes and macrophages, but not on erythrocytes of either larvae or adults. Immunohistochemical observations of the hemopoietic organs revealed that the XL-1+ cells with granulocyte and/or macrophage morphology appeared at st.36-37 in the liver, at st.44-45 in the mesonephric and the thymus rudiments, and at st.47 in the spleen. The XL-1 determinant was expressed on the precursor cells of T lymphocytes in the thymus rudiments at st.46-47, on the pre-B cells in the liver rudiments at st.47, and on lymphocytes in the spleen at st.48-49. A few XL-1+ cells were present in the ventral blood island of the st.35/36 embryos, where differentiating erythrocytes had predominated since st.28. XL-1+ cells with a macrophage-like morphology were found in several locations of the mesenchyme in the st.32 embryos, before the establishment of vascularization at st.33/34 and far earlier than the emergence of lymphocytes.     

Multiple forms of DNA-dependent DNA polymerase during early development and in somatic cells of Xenopus laevis.

Four distinct DNA-dependent DNA polymerase activities (DNA polymerases I, II, III and IV according to the order of elution from a DEAE column) have been separated from extracts of unfertilized Xenopus laevis eggs. The same activities, on the basis of their chromatographic properties, template specificities and sedimentation coefficients, have been found in embryos at least until the gastrula stage. On the other hand, Xenopus kidney cells grown in culture, as well as full grown oocytes lack DNA polymerase I. These data suggest the DNA polymerase I might be a special DNA polymerase activity involved in the extremely rapid DNA synthesis which takes place during early development of X. laevis.     

Distribution of XTdrd6/Xtr protein during oogenesis and early development in Xenopus laevis: Zygotic translation begins only in germ cells that have entered the genital ridge

We previously identified Xenopus tudor domain containing 6/Xenopus tudor repeat (Xtdrd6/Xtr), which was exclusively expressed in the germ cells of adult Xenopus laevis. Western blot analysis showed that the XTdrd6/Xtr protein was translated in St. I/II oocytes and persisted as a maternal factor until the tailbud stage. XTdrd6/Xtr has been reported to be essential for the translation of maternal mRNA involved in oocyte meiosis. In the present study, we examined the distribution of the XTdrd6/Xtr protein during oogenesis and early development, to predict the time point of its action during development. First, we showed that XTdrd6/Xtr is localized to germinal granules in the germplasm by electron microscopy. XTdrd6/Xtr was found to be localized to the origin of the germplasm, the mitochondrial cloud of St. I oocytes, during oogenesis. Notably, XTdrd6/Xtr was also found to be localized around the nuclear membrane of St. I oocytes. This suggests that XTdrd6/Xtr may immediately interact with some mRNAs that emerge from the nucleus and translocate to the mitochondrial cloud. XTdrd6/Xtr was also detected in primordial germ cells and germ cells throughout development. Using transgenic Xenopus expressing XTdrd6/Xtr with a C-terminal FLAG tag produced by homology-directed repair, we found that the zygotic translation of the XTdrd6/Xtr protein began at St. 47/48. As germ cells are surrounded by gonadal somatic cells and are considered to enter a new differentiation stage at this phase, the newly synthesized XTdrd6/Xtr protein may regulate the translation of mRNAs involved in the new steps of germ cell differentiation.