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Biomedical subjects

R Bellairs

Publications and source records attributed to R Bellairs.

51 records · Page 3Linked to original sources

In vivo and in vitro studies on the hypoblast and definitive endoblast of avian embryos.

An unusual example of the invasion of one tissue by another occurs during gastrulation in the chick embryo when the definitive endoblast becomes inserted into the hypoblast. The two tissues were examined morphologically by SEM and TEM. They resemble each other in being of an epithelial type, though neither possesses a basal lamina. The definitive endoblast cells are flatter than the hypoblast cells and more closely attached to one another. When they were explanted in hanging drop cultures, the two tissues were found to exhibit differences in their behaviour. In comparison with the definitive endoblast, the hypoblast cells attached more readily to the glass, produced larger ruffle membranes, moved more rapidly, showed poorer contact-inhibition of locomotion and showed a greater tendency to break away from the main explant. When a hypoblast explant was confronted with a definitive endoblast explant, the hypoblast cells became displaced by the definitive endoblast. The hypoblast explant tended to fragment into smaller groups of cells, many of which migrated around the definitive endoblast, thus mimicking the situation in vivo. Control experiments comprised confronting hypoblast with hypoblast, hypoblast with somites, definitive endoblast with definitive endoblast, and definitive endoblast with somites. The hypoblast explants behaved in a consistent manner, always fragmenting when coming into contact with cells from a confronting explant. The definitive endoblast explants showed more contact inhibition of locomotion when confronted with definitive endoblast or with somites than when confronted with hypoblast. It is suggested therefore that the ability of the hypoblast cells to separate from one another may play an important role in the penetration of the hypoblast by the definitive endoblast both in vitro and in vivo.

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Cell adhesiveness and embryonic differentiation.

The aim of the investigation was to decide whether changes in cell to cell adhesiveness took place during embryonic differentiation. The technique of Curtis (1969) was used to measure the adhesive behaviour of several types of ectodermal, neural and mesodermal cells of the chick embryo at stages 7 and 12 of differentiation. Cells dissected from segmented mesoderm were found to be more adhesive than cells from unsegmented mesoderm. Cells from the ectoderm were more adhesive than those from the neural tissue, at both stages 7 and 12. Cells from both ectoderm and neural tissue became more adhesive between stages 7 and 12. It is concluded that an increase in adhesiveness may play a role in somite segmentation, but not in neural tube formation.

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Placodes of the chick embryo studied by SEM.

The otic, the lens and the nasal placodes have been examined in chick embryos between stages 10 and 18 of Hamburger and Hamilton. At the stage when each placode first becomes visible conspicuous differences have been seen in the surface morphology between those cells which will invaginate and form the placode and those which will remain on the surface of the head, forming the epidermis. The differences become more pronounced with increasing development. The placode cells possess many surface projections whilst the epidermal cells do not. These differences in surface morphology are related to other differences which are visible in TEM sections, the placode cells being highly columnar and extending the full depth of the placode, whilst the epidermal cells are cuboidal or even squamous. This modification in cell shape of the placode cells is correlated with the presence of longitudinally orientated microtubules. The mechanism of invagination is discussed and evidence is presented which supports the idea that there is a migration of cells into the placode from one side. Such a phenomenon would help to explain the asymmetrical structure of the placode, including the presence of the overhanging lip.

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The development of the notochord in the chick embryo, studied by scanning and transmission electron microscopy.

The notochord of the chick embryo between stages 5 and 23 inclusive has been studied by scanning electron microscopy, supplemented by transmission electron microscopy. Three main phases of development are described, and these have been designated: bilaminar; rod-like, unvacuolated; rod-like and vacuolated. The changes in shape of the organ from bilaminar to rod-like is accompanied by changes in the shape, orientation and position of the cells, an increase in the complexity of the cell contacts, and the laying down of a basal lamina. The change from the unvacuolated to the vacuolated phase is accompanied by increasing complexity within the cytoplasm. Most of the vacuoles are intracellular and appear empty though some contain a granular material. The notochordal sheath appears to be secreted by the notochordal cells and fine fibrillar material has been seen in the intercellular spaces. By stage 23, most of the notochordal cells have become so highly vacuolated that the cytoplasm has become closely packed around the nucleus.

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Differentiation of the neural plate and neural tube in the young chick embryo. A study by scanning and transmission electron microscopy.

The differentiation of the presumptive neural plate, the neural plate and the neural tube have been investigated in the chick embryo by SEM, TEM and histochemical techniques. The relationship of these tissues to neighbouring structures, including extracellular materials, has also been studied. When SEM micrographs of primitive streak stage embryos were examined in stereo, it was found that cells which had been invaginating at the time of fixation were similar in shape to fibroblasts migrating in vitro. It was concluded that SEM stereo pairs could provide evidence about the mode and direction of cell migration. Many more mid-bodies have been found associated with the developing neural tissue than with the lateral ectoderm. It was found possible to recognise mid-bodies not only by TEM but also by SEM. It is therefore proposed that SEM montages may be used for assessing which regions of a tissue have recently undergone extensive mitosis. The beads on the specialised threads seen in the early stages of development are now considered to be formed from mid-bodies. Similar, but unbeaded threads have been described which span the gap between the neural folds just prior to the dorsal closure of the neural tube and it seems probably that these threads help to close the neural tube. It is suggested that the beaded threads arise by incomplete separation of two daughter cells at mitosis, whereas the unbeaded threads form by outgrowth of cell processes.

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Freeze-fracture replication of junctionsl complexes in unincubated and incubated chick embryos.

Junctional complexes have been investigated in the epiblast of young chick embryos by examination of freeze-fracture replicas and of sections of comparable specimens stained with lanthanum nitrate. By means of freeze-fracture, tight junctions were shown to be present in the unincubated embryo (stage 1 of Hamburger and Hamilton). The number of ridges or grooves was found to vary between 2 and 10 near the dorsal border, whereas isolated ridges were found more ventrally. Lanthanum was unable to penetrate between the cells in the region of the dorsally situated tight junctions. Similar tight junctions were found in incubated embryos (stage 3) examined by both techniques. Tight junctions were also seen in cleavage (pre-laying) embryos examined in section. Gap junctions were extremely uncommon in unincubated embryos, though occasional aggregates of gap junction particles were seen on the lateral cell membranes close to the dorsal surface. In only one instance were associated pits visible. By contrast, gap junctions were more frequently encountered by stage 3, and these junctions possessed both pits and particles. Desmosomes were never seen in the freeze-fracture replicas at either stages 1 or 3, though structures which might be developing desmosomes were visible in sections. The functions of both the tight and gap junctions in the young chick embryo are discussed. The results are also considered in relation to recent theories about the way in which gap junctions are formed.

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Midbodies and beaded threads.

When the dorsal surface of the young chick embryo is examined by SEM, long threads are visible, each of which appears to connect pairs of cells; these cells may be separated from each other by several intervening cells. Many of the threads possess a bead-like structure about half way along their length. When sections of the beads are examined by TEM they are found to resemble midbodies. Furthermore, the threads possess longitudinally arranged structures within them, which are probably the remnants of the microtubules which were part of a mitotic spindle. It is concluded that each bead is a midbody and that each beaded thread is the remains of a telophase bridge connecting two daughter cells which were incompletely separated after mitosis had taken place. The possible function of the beaded threads is considered.

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The structure of the tapetum of the eye of the sheep.

The structure of the tapetum of the sheep's eye has been examined by transmission electron microscopy. It consists of regularly arranged layers of collagen fibrils, estimated from the hydroxyproline content to make up about 60 percent of the dry weight of the tissue. There are several hundred of these layers in the whole depth, a majority being oriented in the same direction. The collagen is associated with strands of microfibrillar material as well as with cells of both fibrocyte and smooth muscle types.

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