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ABSTRACT: The ultrastructure of the day 8.5 mouse embryo has been studied by transmission electron microscopy, with special emphasis on the primary mesenchymal cells and their interaction with cells of the embryonic ectoderm and the proximal endoderm. The organization of the two polar epithelial cell layers (embryonic ectoderm and proximal endoderm), the isolated cells of the distal endoderm and the primary mesenchymal cells is described. Primary mesenchymal cells are different from embryonic ectoderm cells, from which they are derived, not only by the absence of desmosomes and intermediate-sized filaments of the cytokeratin type but also by their variable morphology not exhibiting stable polar architecture, and their numerous cytoplasmic processes which make contacts with the basal lamina of the ectoderm, the basal cell surface of the proximal endoderm, and other mesenchymal cells. Over most of the embryo the embryonic ectoderm is covered by a typical basal lamina, except for certain regions that are frequently characterized by cytoplasmic projections ("blebs') from the basal cell surface membrane. In contrast, the basal surface of the proximal endoderm is not covered by a continuous basal lamina and reveals mushroom-like protrusions of the cortical cytoplasm. Junctions between primary mesenchymal cells are numerous and include adhaerens-type formations of various sizes as well as gap junctions. Occasionally, a special type of junction between mesenchymal cells and embryonic ectoderm has been found, resulting in local interruptions of the basal lamina. The observations are discussed in relation to possible mechanisms of mesoderm formation and the drastic changes of cell character that accompany this process, including cytoskeletal changes such as the disappearance of cytokeratin filaments and the expression of vimentin.
Differentiation 02/1983; 25(2):121-41. · 2.81 Impact Factor
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ABSTRACT: The cytoskeletal composition of the 'primitive streak' stage of mouse embryos, i.e. at late day 8 ('day 8.5') of gestation, has been examined by electron microscopy, using thin sections of fixed and embedded embryos, and by immunofluorescence microscopy, using cryostat sections of frozen embryos. At this stage primary mesenchymal cells are observed in the posterior part of the embryo which seem to migrate toward the anterior region. For most of the embryo, these mesenchymal cells are separated from the embryonic ectoderm by a continuous basal lamina. Frequently mesenchymal cells form cytoplasmic projections many of which make contact with this basal lamina, with surfaces of proximal endoderm cells, or with other mesenchymal cells. Primary mesenchymal cells contain sparse individual intermediate-sized filaments (IF), but closely packed IF bundles as they occur as tonofibrils in both embryonic epithelia, ectoderm and proximal endoderm, have not been found. Mesenchymal cells also can form junctions of the fascia adhaerens-type but appear to be devoid of desmosomes. Antibodies to cytokeratins reveal strong fibrillar fluorescence in cells of the proximal endoderm and weak, predominantly subapical staining in embryonic ectoderm. Correspondingly, antibodies to desmoplakins, the major proteins of the desmosomal plaque, show punctate fluorescence in both embryonic epithelia. These epithelial cells are not significantly stained with antibodies to other IF proteins such as vimentin and desmin. However, antibodies to vimentin show positive fluorescence, often in fibrillar tangles, in primary mesenchymal cells which in turn are negative with cytokeratin and desmin antibodies. This first detection of expression of vimentin in embryogenesis has been confirmed by two-dimensional gel electrophoresis of cytoskeletal proteins from 35S-methionine-labelled embryos. The observations indicate that during embryogenesis synthesis of vimentin occurs, for the first time, in the primitive streak stage and is restricted to the primary mesenchymal cells. Concomitantly, these cells cease to produce cytokeratins and desmoplakin. Possible mechanisms effective in this rapid change from epithelial to mesenchymal character, i.e. from cytokeratin IF to vimentin IF, are discussed.
Differentiation 02/1982; 23(1):43-59. · 2.81 Impact Factor
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ABSTRACT: Following our study on the expression of cytokeratin filaments in preimplantation mouse embryos [30], we have examined the organization of cytoskeletal elements in early postimplantation embryos up to day 8 of gestation, employing electron microscopy, immunofluorescence microscopy an two-dimensional gel electrophoresis of cytoskeletal proteins labelled by incorporation of 35S-nethionine. The characteristic epithelia formed by the embryonic ectoderm and proximal (visceral) endoderm present well-developed junctional complexes and various differentiated membrane structures. Several apical differentiations of the proximal endodermal cells, such as brush border-like microvilli, the endocytotic labyrinthum, and the supranuclear vacuoles resemble the organization of epithelial cells of the ileum of neonatal mammals. Both embryonic epithelia show typical desmosomes and attached intermediate sized filaments of the cytokeratin type. Other types of intermediate-sized filaments, such as vimentin and desmin filaments, have not been detected in any of the cells of embryos of days 6 and 7, but filaments of the vimentin type can be seen, by immunofluorescence microscopy, late in day 8 in certain cells located in the forming mesoderm. Gel electrophoresis has further revealed that the major cytoskeletal proteins synthesized during days 6-8 in both extraembryonic and embryonic tissue are similar to those characteristic of preimplantation blastocysts and include a major polypeptide corresponding to cytokeratin A described in some internal organs of adult rodents. By the same techniques, synthesis of another cytoskeletal proteins vimentin, has first been found late in day 8. It is concluded that early postimplantation embryonic development, up to mesoderm formation, is characterized by the exclusive presence, in both embryonic ectoderm and proximal endoderm, of differentiated epithelial cells containing desmosome-cytokeratin filament complexes and that other types of intermediate-sized filaments are not yet expressed.
Differentiation 02/1981; 20(3):203-16. · 2.81 Impact Factor
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Differentiation 02/1980; 17(3):161-79. · 2.81 Impact Factor
