Formation of cytoskeletal elements during mouse embryogenesis

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.     

Ultrastructure of the full-term shark yolk sac placenta. II. The smooth, proximal segment

The smooth, proximal portion of the yolk sac placenta of the sandbar shark, Carcharhinus plumbeus is comprised of: (1) An outermost epithelial ectoderm; (2) an intervening collagenous stroma; and (3) an inner mesothelium. The surface epithelium may be one to three cell layers thick. The surface epithelium comprises two cell types. A cuboidal cell that has a dome-like apical surface covered with microvilli and an ovoid nucleus predominate. These cells contain lipid inclusions, many cytoplasmic filaments, and are joined by desmosomes. The second cell type has a convoluted nucleus and a flattened cell apex with microvilli, cilia, and paddle cilia. Golgi complexes and elements of the endoplasmic reticulum are relatively uncommon in the cytoplasm of both cell types. Microplicae also occur on the surface of some cells. The smooth, proximal portion of the placenta is sparsely vascularized. The innermost cellular elements of the surface epithelium rest on a prominent basal lamina. A collagenous zone separates the epithelial basal lamina from the basal lamina of the mesothelium. The mesothelial cells are squamous with a fusiform nucleus, many pinocytotic pits and vesicles, and a large number of cytoplasmic filaments. The endoplasmic reticulum, except for occasional patches of the rough type, and the Golgi complex are poorly developed. Ultrastructural tracer studies show that this portion of the placenta does not absorb horseradish peroxidase (HRP) and trypan blue.     

The role of the basal lamina in mouth formation in the embryo of the starfish Pisaster ochraceus

Details of mouth formation in normal and exogastrulated Pisaster ochraceus larvae have been studied by light microscopy and transmission and scanning electron microscopy. As the archenteron begins to bend, the cells in the presumptive mouth region dissociate and migrate into the blastocoele where they become mesenchyme cells. This leaves a defect in the “blind” endodermal tube, which is covered by a basal lamina. Subsequently this exposed basal lamina bulges to form a blister which appears to extend across the blastocoele to make contact with spikelike projections from the future stomodeal region of the ectoderm. Mesenchyme cell processes are associated with both the basal lamina blister and the ectoderm in this region and may provide both motive power and guidance for contact. Shortly after contact is made the blister of basal lamina from the endoderm fuses with the basal lamina of the ectodermal cells and the ectoderm begins to invaginate. At this time the lateral walls of the presumptive oesophagus are largely formed of naked basal lamina with some loosely associated cells on the endodermal side. Eventually the lateral walls of the proximal part of the oesophagus become cellular, giving rise to an epithelium. A cell plug located between the stomodeum and oesophagus persists for some time before finally breaking down to complete the larval digestive tract. Experiments with exogastrulae suggest that many of these developmental patterns are determined before gastrulation.     

Intermediate filament protein expression and mesoderm formation in the rabbit embryo

Summary This study aims to describe the regulation of vimentin and cytokeratin expression during differentiation of primary mesenchymal cells in the 7 day old rabbit embryo; unusual intermediate filament protein expression patterns have already been found in this species at later embryonic stages. Double-labelling indirect immunofluorescence assays with a panel of monoclonal intermediate filament antibodies are performed on frozen sections and compared with aldehyde-fixed plastic-embedded tissues. The histological part of the study, serving as a basis for the topographical orientation in the immunostained frozen sections, emphasises many similarities between the primitive streak embryos of the rabbit and the chick. The immunohistochemical analysis reveals cytokeratin expression to varying degrees in all germ layers. Vimentin expression, always in combination with cytokeratin expression, is found in a few cells of the ectoderm, endoderm and lateral mesoderm, but not in the primary mesenchymal cells of either the primitive node or the primitive streak. The results are discussed in relation to recent experimental findings on differentiation and morphogenetic processes in the primitive streak embryo. While these complex expression patterns make it seem unlikely that intermediate filament protein subtypes are expressed independently of cellular function during development, no indication can be found for a relation between vimentin expression and the morphogenetic changes thought to be important during mesoderm formation.Supported by the Deutsche Forschungsgemeinschaft (Wa 359-9) and by the Netherlands Cancer Foundation Offprint requests to: C. Viebahn     

Three cell recognition changes accompany the ingression of sea urchin primary mesenchyme cells

At gastrulation the primary mesenchyme cells of sea urchin embryos lose contact with the extracellular hyaline layer and with neighboring blastomeres as they pass through the basal lamina and enter the blastocoel. This delamination process was examined using a cell-binding assay to follow changes in affinities between mesenchyme cells and their three substrates: hyalin, early gastrula cells, and basal lamina. Sixteen-cell-stage micromeres (the precursors of primary mesenchyme cells), and mesenchyme cells obtained from mesenchyme-blastula-stage embryos were used in conjunction with micromeres raised in culture to intermediate ages. The micromeres exhibited an affinity for hyalin, but the affinity was lost at the time of mesenchyme ingression in vivo. Similarly, micromeres had an affinity for monolayers of gastrula cells but the older mesenchyme cells lost much of their cell-to-cell affinity. Presumptive ectoderm and endoderm cells tested against the gastrula monolayers showed no decrease in binding over the same time interval. When micromeres and primary mesenchyme cells were tested against basal lamina preparations, there was an increase in affinity that was associated with developmental time. Presumptive ectoderm and endoderm cells showed no change in affinity over the same interval. Binding measurements using isolated basal laminar components identified fibronectin as one molecule for which the wandering primary mesenchyme cells acquired a specific affinity. The data indicate that as the presumptive mesenchyme cells leave the vegetal plate of the embryo they lose affinities for hyalin and for neighboring cells, and gain an affinity for fibronectin associated with the basal lamina and extracellular matrix that lines the blastocoel.     

Cytoskeleton and thyroglobulin expression change during transformation of thyroid epithelium to mesenchyme-like cells

In considering the mechanism of transformation of epithelium to mesenchyme in the embryo, it is generally assumed that the ability to give rise to fibroblast-like cells is lost as epithelia mature. We reported previously that a definitive embryonic epithelium, that of the anterior lens, gives rise to freely migrating mesenchyme-like cells when suspended in type I collagen matrices. Here, we show that a highly differentiated epithelium that expresses cytokeratin changes to a vimentin cytoskeleton and loses thyroglobulin during epithelial-mesenchymal transformation induced by suspension in collagen gel. Using dispase and collagenase, we isolated adult thyroid follicles devoid of basal lamina and mesenchyme, and we suspended the follicles in 3D collagen gels. Cells bordering the follicle lumen retain epithelial polarity and thyroid phenotype, but basal cell surface organization is soon modified as a result of tissue multilayering and elongation of basal cells into the collagenous matrix. Cytodifferentiation, determined by thyroglobulin immunoreactivity, is lost as the basal epithelial cells move into the matrix after 3-4 days in collagen. By TEM, it can be seen that the elongating cells acquire pseudopodia, filopodia and mesenchyme-like nuclei and RER. Immunofluorescence examination of intermediate filaments showed that freshly isolated follicles and follicles cultured on planar substrata react only with anticytokeratin. However, all of the mesenchyme-like cells express vimentin and they gradually lose cytokeratin. These results suggest that vimentin may be necessary for cell functions associated with migration within a 3D matrix. The mesenchymal cells do not revert to epithelium when grown on planar substrata and the transformation of epithelium to mesenchyme-like cells does not occur within basement membrane gels. The results are relevant to our understanding of the initiation of epithelial-mesenchymal transformation in the embryo and the genetic mechanisms controlling cell shape, polarity and cytoskeletal phenotype.     

Changing patterns of cytokeratins and vimentin in the early chick embryo

The distribution of cytokeratins and vimentin intermediate filaments in the first 48 h of chick development has been determined using immunofluorescent labelling. During formation of the germ layers, cytokeratin expression is associated with the appearance of an integral epithelium (ectoderm), whereas vimentin expression is associated with cells that detach and migrate from this epithelium to form endoderm and mesoderm. Subsequently, vimentin persists in the endoderm and mesoderm and the tissues derived therefrom, such as the somites and developing heart, throughout the period of study. The appearance of cytokeratins at later stages of development occurs in some epithelia such as the ectoderm, endoderm, lateral plate and epimyocardium but not others including the neural plate, neural tube and somites. Expression of cytokeratins in endoderm and mesenchymal tissues occurs in tandem with vimentin. In conclusion, vimentin expression is related to its distribution in the epiblast before germ layer formation. Its initial appearance may be related to the motile behaviour of cells about to ingress through the primitive streak. The appearance of cytokeratin filaments, however, does not reflect germ layer derivation but rather the need for an epithelial sheet.     

The structure and activities of echinonectin: a developmentally regulated cell adhesion glycoprotein with galactose-specific lectin activity.

The extracellular matrix of the sea urchin embryo contains a 230 kD homodimeric glycoprotein known as echinonectin (EN). EN contains a cell attachment domain as well as a galactose-specific lectin activity. Cell attachment to EN is differentially regulated in the three primary germ layers, endoderm, ectoderm and mesoderm. Prior to gastrulation all embryonic cells adhere equally to EN-coated substrates, but during gastrulation primary mesenchyme cells lose affinity for EN, ectoderm cells increase their binding to the molecule, and cells of the endoderm maintain a similar or slightly lowered level of binding. The mechanisms governing these adhesive changes and the specific functions they serve in development are not currently understood. They are timed to coincide with distinct morphogenetic events such as primary mesenchyme cell ingression and archenteron formation, suggesting that regulated adhesion to EN plays at least a permissive role in early morphogenesis.     

Expression of syndecan, a putative low affinity fibroblast growth factor receptor, in the early mouse embryo.

Syndecan is an integral membrane proteoglycan that binds cells to several interstitial extracellular matrix components and binds to basic fibroblast-growth factor (bFGF) thus promoting bFGF association with its high-affinity receptor. We find that syndecan expression undergoes striking spatial and temporal changes during the period from the early cleavage through the late gastrula stages in the mouse embryo. Syndecan is detected initially at the 4-cell stage. Between the 4-cell and late morula stages, syndecan is present intracellularly and on the external surfaces of the blastomeres but is absent from regions of cell-cell contact. At the blastocyst stage, syndecan is first detected at cell-cell boundaries throughout the embryo and then, at the time of endoderm segregation, becomes restricted to the first site of matrix accumulation within the embryo, the interface between the primitive ectoderm and primitive endoderm. During gastrulation, syndecan is distributed uniformly on the basolateral cell surfaces of the embryonic ectoderm and definitive embryonic endoderm, but is expressed with an anteroposterior asymmetry on the surface of embryonic mesoderm cells, suggesting that it contributes to the process of mesoderm specification. In the extraembryonic region, syndecan is not detectable on most cells of the central core of the ectoplacental cone, but is strongly expressed by cells undergoing trophoblast giant cell differentiation and remains prominent on differentiated giant cells, suggesting a role in placental development. Immunoprecipitation studies indicate that the size of the syndecan core protein, although larger than that found in adult tissues (75 versus 69 x 10(3) Mr), does not change during peri-implantation development. The size distribution of the intact proteoglycan does change, however, indicating developmental alterations in its glycosaminoglycan composition. These results indicate potential roles for syndecan in epithelial organization of the embryonic ectoderm, in differential axial patterning of the embryonic mesoderm and in trophoblast giant cell function.     

Endo16 is required for gastrulation in the sea urchin Lytechinus variegatus

The Endo16 gene encodes a large extracellular protein with several functional domains that provide some insight into the role of this protein during embryonic development. We isolated the full-length cDNA sequence from Lytechinus variegatus and utilized morpholinos to further investigate the role of Endo16 during embryonic development in this species. Endo16-deficient embryos failed to undergo gastrulation and the blastocoele became filled with dissociated cells after 24 h of incubation. Moreover, there was a delay in endoderm differentiation as assayed by staining with an antibody that recognizes Endo1. The differentiation of other cell types including oral ectoderm, primary mesenchymal cells (PMC) and secondary mesenchymal cells (SMC) appeared to be normal, although the patterns of protein expression did not resemble control embryos due to the gross morphological abnormalities elicited by the LvEndo16 morpholino. Microinjection of full-length EGFP mRNA with the LvEndo16 morpholino-targeted sequence confirmed that this phenotype can be attributed specifically to the loss of Endo16 protein. Taken together, our data suggest that Endo16 may be required for the cell-extracellular matrix (ECM) interactions that are required for endoderm differentiation in the sea urchin embryo.     

An ultrastructural analysis of cell and matrix differentiation during early limb development in Xenopus laevis

Early development of the hind limb of Xenopus (stages 44–48) has been analyzed at the level of ultrastructure with emphasis on differentiation of extracellular matrix components and intercellular contacts. By stages 44–45, mesenchyme is separated from prospective bud epithelium by numerous adepidermal granules in a subepithelial compartment (the lamina lucida), a continuous basal lamina and several layers of collagen (the basement lamella). Tricomplex stabilization of amphoteric phospholipid demonstrates that each adepidermal granule consists of several membranelike layers (electron-lucent band 25–30 Å; electron-dense band 20–40 Å), which are usually parallel to the basal surface of adjacent epithelial cells. Collagen fibrils are interconnected by filaments (35 Å in diameter) which stain with ruthenium red. Epithelial cells possess junctional complexes at their superficial borders, numerous desmosomes at apposing cell membranes and hemidesmosomes at their basal surface. Mesenchymal cells predominantly exhibit close contacts (100–150 Å separation) with few focal tight junctions at various areas of their surface. By stages 47–48, adepidermal granules are absent beneath bud epithelium and layers of collagen in the basement lamella lose filamentous cross-linking elements. Filopodia of mesenchymal cells penetrate the disorganized matrix and abut the basal lamina. Hemidesmosomes disappear at the basal surface of the epidermis and mesenchymal cells immediately subjacent to epithelium exhibit focal tight junctions and gap junctions at their lateral borders. These structural changes may be instrumental in the epitheliomesenchymal interactions of early limb development. Degradation of oriented collagenous lamellae permits direct association of mesenchymal cell surfaces (filopodia) with surface-associated products of epithelial cells (organized into the basal lamina). Development of structural pathways for intercellular ion and metabolite transport in mesenchyme may coordinate events specific to limb morphogenesis.     

Ultrastructure and synthesis of the extracellular matrix of Pisaster ochraceus embryos preserved by freeze substitution

When asteroid embryos cryoprotected with propylene glycol are rapidly frozen in liquid propane and freeze substituted with ethanol, preservation of the cells and extracellular matrix (ECM) is excellent. The basal lamina, although thicker and less well defined than in conventionally fixed embryos, demonstrates a region of decreased density just below the cells that corresponds to the lamina lucida and a lamina densa. The former region is often occupied by fibrous material. In addition, as was previously described in conventionally fixed tissues, the basal lamina of the ectoderm is generally thicker and more substantial than that of the endoderm, reinforcing an earlier suggestion that the structure of the basal lamina is different in different regions of the embryo. The ECM of the blastocoel consists of thin “twig-like” elements that form a loose meshwork evenly distributed throughout the blastocoel. Bundles of 20 nm fibers, located within the meshwork, are oriented parallel to the base of the cells of the stomodeum. In the long axis of the embryo, similar fibers are present in the dorsal aspect of the animal between the stomach and the ectoderm and radiate out from the esophagus crossing the region between it and the ectoderm. Immunocytochemical work with three different monoclonal antibodies shows that glycoprotein molecules, synthesized in the Golgi apparatus, are also secreted here and form part of the matrix structure. The results suggest that the blastocoel is filled with a gel-like material reinforced with bundles of 20-nm fibers. The manner in which the observed arrangement could contribute to the development and maintainence of the shape of the embryo is discussed. J Morphol 232:133–153, 1997. © 1997 Wiley-Liss, Inc.     

Development of the notochord in normal and malformed human embryos and fetuses.

In view of its possible involvement in early embryogenesis and teratogenesis, the developmental characteristics of the human notochord were studied by light and electron microscopy and immunohistochemistry on 20 human conceptuses (5th-22nd week). At the earliest embryonic stages examined, the notochord is closely related to both the pharyngeal endoderm and the neuroectoderm of the posterior (tail) end of the neural tube. In both regions the interspace is bridged by slender cytoplasmic processes, lined with basal lamina and filled with amorphous extracellular material containing collagen types III and IV and laminin. The notochordal cells express cytokeratin brightly and vimentin weakly. As embryonic age progresses, the notochord gradually separates from the epithelia, becomes the axis of developing spinal column and undergoes progressive cell degeneration and rearrangement within the vertebral bodies. This is associated with extensive production of extracellular material and the first appearance of fibronectin. Intracellularly, the expression of vimentin gradually increases, while that of cytokeratin slightly weakens. Changes in the notochord parallel other developmental events in axial organs. In anencephalic fetuses the course of the notochord is irregular and partly interrupted with segments outside the basichondrocranium in specimens with craniorachischisis.     

Morphological changes in the oral (buccopharyngeal) membrane in urodelan embryos: development of the mouth opening

The ultrastructure of the oral (buccopharyngeal) membrane in the embryo of the urodelan, Hynobius tokyoensis, was examined by transmission (TEM) and scanning electron microscopy (SEM). The oral membrane consists of the stomodeal ectoderm and foregut endoderm, and is three to five cell layers thick at stage 24. The oral membrane gradually thickens as development proceeds. The stomodeal collar, derived from the ectoderm, is folded inward along the foregut endoderm. Tooth germs are formed partly by cells of the stomodeal collar and partly by mesenchymal cells and calcification takes place before hatching. Secretory granules, which are markers of epithelial differentiation, appear in some cells of the foregut endoderm. Within the oral membrane, the cells of the stomodeal collar become the basal cells, and the endodermal cells of the foregut become the apical cells of the future oral epithelium. Gaps are formed by the epithelial differentiation of the endodermal cells of the foregut in the oral membrane. The gaps connect with each other, with the stomodeum, and with the foregut. As a result of these events, the mouth opens at stage 43, just after hatching.     

Fine structure of the rhombencephalic tela of the bullfrog,Rana catesbeiana

Summary The posterior rhombencephalic tela choroidea of the bullfrog was examined by electron microscopy. This membrane, the pia-ependymal roof of the caudal hindbrain, contains a large central region characterized by cuboidal ependymal cells which surround sizable microscopic apertures — the interependymal pores.Ultrastructurally ependymal cells of this area are characterized by infrequent apical microvilli and cilia. They contain irregularly shaped nuclei and few cytoplasmic organelles that are largely apical in position. The most striking feature is an abundance of cytoplasmic filaments forming an extensive cytoskeleton. Laterally these cells are joined by numerous elaborate desmosomes. The majority of the ependymal cells have a basal lamina consisting of single, double, or triple laminae lying parallel to the basal plasma membrane.Several unusual specializations are seen at the margins of the interependymal pores. The ependymal cells have lateral cytoplasmic processes that form the actual border of each pore. These processes originate from the apical surface of the cell and partially enclose an elaborate network of basal lamina associated with the interependymal pores.These findings demonstrate microscopic apertures in the roof of the fourth ventricle in the bullfrog that are associated with an unusual form of supportive ependyma.     

Palatogenesis in hamster. II. Ultrastructural observations on the closure of palate

Formation of secondary palate in hamster was studied with electron microscopy. Prior to assuming horizontal position, the palatal shelves were covered by a two to three cell layer thick epithelium which was separated from the underlying mesenchyme by an intact basal lamina. Epithelial cells were attached to each other by desmosomes. Early hemidesmosomes could be identified as thickenings of the cytoplasmic membrane opposing the basal lamina. Epithelial cells, like other embryonic cells, contained only few organelles but were rich in polyribosomes. As the horizontal shelves approached each other towards the midline, lysosomes and tonofilaments appeared in the superficial and basal cells of the epithelia. Superficial cells showed degeneration and eventual lysis. Fusion of the opposing epithelia occurred between the deeper cells by means of newly formed desmosomes. The epithelial seam resulting from fusion of the epithelia was limited on each side by a continuous basal lamina. Its subsequent thining and eventual fragmentation resulted from the loss of cells by autophagy. There was no evidence of mesenchymal invasion of the epithelial seam. Mesenchymal macrophages appeared in the later stage of palatogenesis and were responsible for phagocytosis of cellular debris. Formation of the soft palate was basically similar to that of the secondary hard palate and occurred by fusion of the opposing shelves. Similarly, anterior closure of the palate occurred by fusion of the lower end of the nasal septum to the primary and secondary palates. Hyperplasia of the opposing epithelia, prior to their fusion, was often seen. It is suggested that formation of the palate occurs in predictable and coordinated fashion and that timely appearance of lysosomes causing lysis of intervening epithelia is of great significance in normal palatogenesis.     

Ultrastructure of the pre-implantation shark yolk sac placenta

During ontogeny, the yolk sac of viviparous sharks differentiates into a yolk sac placenta which functions in gas exchange and hematrophic nutrient transport. The pre-implantation yolk sac functions in respiration and yolk absorption. In a 10.0 cm embryo, the yolk sac consists of six layers, viz. (1) somatic ectoderm; (2) somatic mesoderm; (3) extraembryonic coelom; (4) capillaries; (5) endoderm; and (6) yolk syncytium. The epithelial ectoderm is a simple cuboidal epithelium possessing the normal complement of cytoplasmic organelles. The endoplasmic cisternae are dilated and vesicular. The epithelium rests upon a basal lamina below which is a collagenous stroma that contains dense bodies of varying diameter. They have a dense marginal zone, a less dense core, and a dense center. The squamous mesoderm has many pinocytotic caveolae. The capillary endothelium is adjacent to the mesoderm and is delimited by a basal lamina. The endoderm contains yolk degradation vesicles whose contents range from pale to dense. The yolk syncytium contains many morphologically diverse yolk granules in all phases of degradation. Concentric membrane lamellae form around yolk bodies as the main yolk granules begin to be degraded. During degradation, yolk platelets exhibit a vesicular configuration.     

Ultrastructural Peculiarities of the Embryogenesis of the Brittle Star Amphipholis kochii (Lutken, 1972)

This paper addresses morphogenetic processes and cell differentiation during embryogenesis of the brittle star Amphipholis kochii at the ultrastructural level. The radial cleavage is not strictly determined. Embryos are covered with a thick hyaline envelope and contain numerous yolk granules and small lipid drops. Blastulae feature a thick blastoderm with extensive intercellular cavities, which are retained in the crest epithelium of late gastrulae. Embryonic cells have single cilia with long cross-striated rootlets associated with the Golgi apparatus. Depolarized cells of the primary mesenchyme with a well-developed rough endoplasmic reticulum differentiate into sclerenchyme syncytium. Gastrulation occurs by invagination. Secondary mesenchymal cells emigrate from the archenteron tip to differentiate into amebocytes, which contain a well-developed Golgi apparatus and numerous mitochondria. The endoderm is formed of cubic cells with numerous yolk granules and rare microvilli. Flattened cells of the dorsal and ventral ectoderm contain a small amount of yolk. Yolk utilization during embryogenesis occurs by intracellular lysosomal digestion with selective exocytosis of toposomes.Original Russian Text Copyright © 2005 by Biologiya Morya, Gliznutsa, Dautov.     

An electron microscopic analysis of notochordal and mesenchymal cell abnormalities in embryos of Danforth’s short-tail (Sd) mice

Abnormalities of notochordal cells and of mesenchymal cells in embryos of Danforth's short-tail (Sd) and C57BL mice were examined by means of electron microscopy and cytochemical staining at 11.0 and 11.5 days of gestation. In abnormal (Sd/+; Sd/Sd) embryos, the notochordal cells were markedly deficient in bundles of filaments and lacked surface protrusions, and the notochordal basal lamina was continuous; in contrast, notochordal cells of normal (+/+) littermates and of C57BL embryos contained numerous bundles of filaments and showed fingerlike surface protrusions and discontinuous basal laminae. The pathologic notochordal cells also lacked the accumulations of glycogen revealed in the normals by means of thiocarbohydrazide cytochemical staining at the electron microscopic level. The mesenchymal cells of abnormals also were deficient in filaments but did stain for glycogen, though less prominently than did normal mesenchymal cells.     

Parietal and visceral endoderm differ in their expression of intermediate filaments

Two layers of extra-embryonic endoderm, viz. the parietal endoderm (PE) and the visceral endoderm (VE), arise in the mouse embryo shortly after implantation. Both cell populations apparently originate from the primitive endoderm of the blastocyst. While the endoderm differentiation has been studied both in the embryo and in the embryonal carcinoma model system, the investigation has been hampered by the paucity of unequivocal markers of differentiation, especially in the case of the PE. Here we show that the PE and VE of mouse conceptuses differ in their expression of intermediate filaments: while both cell types contain cytokeratin, expression of vimentin was only revealed in the cells of the PE. The association between the differentiation of PE and the appearance of vimentin filaments is discussed.