Expression of XNOA 36 in the mitochondrial cloud of Xenopus laevis oocytes

SummaryIn Xenopus laevis oocytes a mitochondrial cloud (MC) is found between the nucleus and the plasma membrane at stages I–II of oogenesis. The MC contains RNAs that are transported to the future vegetal pole at stage II of oogenesis. In particular, germinal plasm mRNAs are found in the Message Transport Organiser (METRO) region, the MC region opposite to the nucleus. At stages II–III, a second pathway transports Vg1 and VegT mRNAs to the area where the MC content merges with the vegetal cortex. Microtubules become polarized at the sites of migration of Vg1 and VegT mRNAs through an unknown signalling mechanism. In early meiotic stages, the centrioles are almost completely lost with their remnants being dispersed into the cytoplasm and the MC, which may contain a MTOC to be used in the later localization pathway of the mRNAs. In mammals, XNOA 36 encodes a member of a highly conserved protein family and localises to the nucleolus or in the centromeres. In the Xenopus late stage I oocyte, XNOA 36 mRNA is transiently segregated in one half of the oocyte, anchored by a cytoskeletal network that contains spectrin. Here we found that XNOA 36 transcript also localises to the nucleoli and in the METRO region. XNOA 36 protein immunolocalization, using an antibody employed for the library immunoscreening that depicted XNOA 36 expression colonies, labels the migrating MC, the cytoplasm of stage I oocytes and in particular the vegetal cortex facing the MC. The possible role of XNOA 36 in mRNA anchoring to the vegetal cortex or in participating in early microtubule reorganization is discussed.

The migration of presumptive primordial germ cells through the endodermal cell mass in Xenopus laevis: A light and electron microscopic study

Presumptive primordial germ cells (pPGCs) were examined during migration from their deep endodermal position to the endodermal crest in Xenopus laevis, using light and electron microscopy with Epon sections, and several morphological characteristics of pPGCs, associated with their migration, were revealed. pPGCs displayed polymorphism, with smooth contours. The intercellular space around the pPGCs was large and variable in width and cytoplasmic processes from pPGCs were occasionally observed in it. It was shown quantitatively that pPGCs at the migratory stage had a tendency to move with the leading end, towards which the nucleus was localized, dragging the germinal plasm behind. These polarized pPGCs were frequently associated with large intercellular spaces, both at their leading and trailing ends. Cytoplasmic processes of polarizing pPGCs found in the large intercellular space at the leading end were conspicuous. Ultrastructurally, the nuclei of pPGCs were euchromatic, and the nucleolus was prominent. The germinal plasm at the light microscope level corresponded to the cytoplasmic area near the nucleus where a large number of mitochondria with well-developed cristae and most of the other organelles were aggregated. Centrioles and centriole-associated microtubules observed in the aggregate were thought to be important structures responsible for the cell polarization mentioned above. It was demonstrated quantitatively that the size of mitochondria in pPGCs was larger on average than that of mitochondria in neighbouring somatic endodermal cells. Numerous irregularly shaped small yolk platelets characterized pPGCs. These ultrastructural features suggested that pPGCs were in an activated metabolic state. It was concluded that the migration of pPGCs was attributable to active movement with high cell metabolism, causing the formation of cell processes and intracellular polarization.

Relationship between the amount of the ‘germinal plasm’ and the number of primordial germ cells in Xenopus laevis

Tadpoles of Xenopus laevis completely lacking primordial germ cells were obtained by irradiating the vegetal hemisphere of early 2-cell eggs with u.v. (wavelength, 253·7 nm; dose, ca. 6000 ergs/mm2). An increasing number of primordial germ cells were observed as the stage at irradiation advanced from early 2-cell to early 4-cell stages. Furthermore, early 2-cell eggs irradiated with doses ranging from 750 to 6000 ergs/mm2 grew into tadpoles carrying a decreasing number of primordial germ cells in accord with the increase of the dose. On the other hand, tadpoles developed from eggs irradiated immediately after being centrifuged at 150 g for 1 min at early 2-cell stage to displace the ‘germinal plasm’ deeper into the cytoplasm, carried a considerable number of primordial germ cells. These facts were interpreted to suggest the presence of u.v.-sensitive germ cell determinant in the ‘germinal plasm’. It was revealed by varying the area of irradiation that the number of primordial germ cells decreased in direct proportion to the increase of the area irradiated. It was concluded that the amount of the u.v.-sensitive material(s) contained in the ‘germinal plasm’ determined the number of primordial germ cells in tadpoles.