Search PubMed⌕ Search

Biomedical subjects

R Bellairs

Publications and source records attributed to R Bellairs.

At least 37 records · Page 2Linked to original sources
PubMed1985

Isolation, characterization and localization of a lectin within the vitelline membrane of the hen's egg.

A lectin with an affinity for certain sulphated polysaccharides, such as fucoidin and dextran sulphate, has been isolated from the vitelline membrane of hens' eggs and purified to homogeneity as assessed by two-dimensional gel electrophoresis. Polyclonal and monoclonal antibodies have been raised to the lectin and used in indirect immunofluorescence microscopy to localize the agglutinin in the outer layer of the vitelline membrane, where the lectin persists prior to the breakdown of the vitelline membrane. The quantity of lectin extracted from the two layers of the membrane, which have been separated by the method of Bellairs, Harkness & Harkness (1963), correlated well with the results of immunofluorescence microscopy. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of the two layers of the membrane indicates that each layer has a distinctive polypeptide composition, the outer layer containing in particular lysozyme and avidin. The evidence obtained in this study indicates that the lectin is not involved in adhesion of the blastoderm to the vitelline membrane; neither is it involved in the expression of the blastoderm nor in maintaining the strength of the membrane. The possible roles in promoting transport of solutes across the membrane as well as providing bactericidal properties to the egg are discussed.

Animals↗
PubMed1985

Protein synthetic patterns of tissues in the early chick embryo.

Tissues dissected from early chick embryos were labelled in vitro with [35S]methionine, and their patterns of polypeptide synthesis investigated using the technique of two-dimensional (2-D) polyacrylamide gel electrophoresis. Apart from providing a preliminary description of the molecular changes associated with the processes of gastrulation and segmentation in the chick embryo, this study has revealed a number of polypeptides that may be useful as markers of cell type or function. The protein synthetic patterns of hypoblast from early and late gastrulae (stages 2 and 4, respectively: Hamburger & Hamilton, 1951) and of definitive endoblast and junctional endoblast from late gastrulae all resemble one another closely, but differ markedly from that of the epiblast at either stage. The lower layer tissues are characterized by the presence of eleven polypeptides that are largely absent from the epiblast. These findings are discussed with reference to current theories on the origins of the lower layer tissues. Comparisons between the 2-D patterns for tissues dissected from gastrulae and from embryos undergoing segmentation (stage 12) have revealed ten polypeptides showing stage-specific rather than tissue-specific expression. Apart from these ten polypeptides, the 2-D patterns for epiblast and ectoderm were practically identical, and distinguishable from those of other tissues by a lack of any unique polypeptides. On the other hand, stage-4 endoblast and stage-12 endoderm differed in the expression of many polypeptides. One polypeptide was found that may be considered as a marker of mesodermal cell type, as it was present in lateral plate, segmental plate and somitic mesoderm, but not in tissues of the other germ layers. Lateral plate could be distinguished from the other mesodermal tissues in the expression of a number of polypeptides, but the similarity in the 2-D patterns for segmental plate and somites suggest that the separation of somites from the anterior end of the segmental plate is not accompanied by the synthesis of new polypeptides.

Animals↗
PubMed1984

Mitotic activity during somite segmentation in the early chick embryo.

The mitotic activity of the somites, segmental plate and posterior mesoderm were investigated in colchicine-treated and untreated chick embryos at st. 7-14. The mitotic figures in the somites are restricted to the proximity of the lumen and have their spindles orientated predominantly tangentially to the cavity. In the segmental plate there is no pattern in terms of the position or orientation of the mitotic spindles, but there is a single region, often found close to the cranial end of the segmental plate, with an elevated mitotic index. This may indicate a certain degree of synchrony among groups of segmental plate cells. These results are discussed in relation to the process of somite segmentation.

Animals↗
PubMed1984

Effect of microtubule inhibitors on the expansion of hypoblast and margin of overgrowth of chick blastoderms.

The effect of the microtubule inhibitors, Nocodazole and taxol, was studied on the expansion of fragments of chick hypoblast (8 to 10 h incubation) and of margin of overgrowth (24 h incubation) cultured on artificial substrata and on the epiboly in intact blastoderms (24 h incubation). Immunocytochemical staining of these cells with antiserum against tubulin showed that 1 micrograms Nocodazole/ml caused disassembly of microtubules, and that 1 microgram taxol/ml led to increased but unordered assembly. The solvent dimethylsulphoxide had no effect. At these concentrations both microtubule inhibitors led to rapid arrest of the expansion of fragments of hypoblast and of margin of overgrowth in culture, and of the epiboly in intact blastoderms. Time-lapse films showed that inhibition of expansion in both situations was reversible within 2 h after removal of the drugs. Phase-contrast microscopy showed remarkably little difference between the morphology of treated as compared to untreated cultures. Measurements of the height of the cells on sectioned fragments of margin of overgrowth showed no differences between treated and untreated cultures. These results suggest that the cytoplasmic microtubule complex is important both for epiboly and for the migration of hypoblast cells in the chick blastoderm. The mechanisms of this microtubule-related migration are not understood.

Alkaloids↗
PubMed1984

The roles of node regression and elongation of the area pellucida in the formation of somites in avian embryos.

Experiments have been carried out on explanted chick embryos to test certain widely accepted concepts about the role of Hensen's node in somite formation. The relationship between elongation of the area pellucida and regression of Hensen's node has also been investigated. We conclude from these experiments that: (a) The timing of somite formation is not controlled by the regression of Hensen's node, nor by the shearing of the mesoderm into right and left halves. (b) Somite size and shape are probably controlled by local conditions in the chick embryo. (c) Elongation and regression are two different events. (d) The position of the probably depends on mechanical tensions in the area pellucida. (e) The notochord is not required for the stability of somites in vivo.

Agar↗
PubMed1984

Experimental analysis of control mechanisms in somite segmentation in avian embryos. II. Reduction of material in the gastrula stages of the chick.

A new theory of control of somite segmentation in chick embryos is proposed. This supposses that tiny clusters of already programmed cells are present throughout the presumptive somite area at stage 4, but that in order to fulfill their destiny they probably depend on the addition of further cells from the primitive streak. Evidence is based on the two groups of experiments: a) Experiments involving transection across the primitive streak at various stages, (which results in a 'tail' which possesses mesodermal derivatives) and across the segmental plate (which results in a 'tail' lacking mesodermal derivatives). b) Experiments in which parts of embryos have been explanted with or without their primitive streak. It is suggested that the initial clusters of pre-programmed cells move further and further posteriorly, developing into somitomeres (the precursors of true somites) only as they receive re-inforcements from the primitive streak or, ultimately, from the tail bud.

Animals↗
PubMed1983

Interaction of three human malignant cell lines with chick hypoblast in culture.

The hypoblast (lower layer) was dissected from young chick blastoderms and explanted in vitro, where it formed an epitheloid sheet. Cells from the following malignant lines were explanted on top of the sheet both as aggregates and as cell suspensions: Hu456 human bladder carcinoma, SAOS-2 human osteosarcoma, LICR(LOND)-HN-4 laryngeal carcinoma. The interaction of the malignant cells with the hypoblast was studied by time lapse cinephotography, light microscopy, and transmission electron microscopy. All malignant cells penetrated through the hypoblast, so that a gradually enlarging hole formed in it. Apart from this common pattern of behaviour, the three types of malignant cells differed in their interactions with the hypoblast in the following ways. 1) Both the Hu456 and to a lesser extent the SAOS-2 cells brought about an initial retraction of the hypoblast so that a temporary cell-free space was formed. No such retraction occurred in response to the LICR-(LOND)-HN-4 cells. 2) Each of the three types of malignant cells migrated for some distance beneath the hypoblast, and in this area of underlap, there were differences in the amount and disposition of extracellular material. Thus, there was more extracellular material between the hypoblast and underlying SAOS-2 cells than between the hypoblast and underlying Hu456 cells, whilst there was no extracellular material between the hypoblast and underlying LICR(LOND)-HN-4 cells. Indeed, the hypoblast and LICR(LOND)-HN-4 cells often shared desmosomes. 3) When explanted as aggregates on hypoblast Hu456 and SAOS-2 cells left the corona and migrated as solitary cells underneath the hypoblast in contrast with control aggregates explanted on plastic. These cells which had migrated beneath the hypoblast were flatter than their corresponding control cells which had spread on the plastic substrate. The flatter cells appeared to have been using the extracellular materials as a substrate, rather than the plastic. Such differences in the migratory behaviour between experimental and control cultures were not observed with LICR(LOND)-HN-4 cells.

Animals↗
PubMed1983

Experimental analysis of control mechanisms in somite segmentation in avian embryos. I. Reduction of material at the blastula stage in Coturnix coturnix japonica.

The blastulae of unincubated eggs of the quail, Coturnix coturnix japonica, have been bisected in ovo, using the technique of Lutz (1949). Some embryos were harvested after 24 h and found to possess two primitive streaks. Most were fixed at 48 h or 72 h. Some were found to have regulated to form almost normal single axes, whilst others had developed into duplicitas anterior embryos, separate twins or collided axes. All three types of twinned embryos were smaller than the control embryos. The number of somites was not however reduced in the shorter embryos. This finding corresponds with a similar result obtained by Cooke (1975) who reported that if a Xenopus blastula is reduced in size, it nevertheless develops the correct number of somites. The quail however adjusts the shape of the individual somites so that they fit into the reduced body length, whereas Xenopus reduces the size of somites. No miniaturized somites were ever seen in these quail embryos. As a result of the present experiments, it was concluded that the length of incubation time does not directly control the rate of somite formation, because different numbers of somites were found in twins which possessed identical genomes and had developed in almost the same environment for identical periods. In addition, the size of the area pellucida does not appear to control somite formation. Probably, the most important influence is the regression of the node.

Animals↗
PubMed1982

An electron-microscopical analysis of embryonic chick tissues explanted in culture.

Three types of tissue (hypoblast, germ wall and epiblast) were dissected from early chick embryos and explanted on Falcon plastic dishes. After they had settled and spread, the explants were fixed, usually within 18-24 h after explantation, and sections were cut through the tissue and the Falcon dish. The closeness of the cells to the substrate varied even within the same explant, but the epiblast tended to be closer to the substrate than did the hypoblast or germ wall. Plaques were present in all three tissues in regions where the cell processes contracted the substrate. Extensive desmosomes were visible in the epiblast explants, small desmosomes were present in the germ wall explants, but desmosomes were never seen in hypoblast explants. These differences in cell/substrate and cell/cell morphology are discussed in relation to the different behavioural characteristics of the three tissues. Some mixed cultures were also examined by electron microscopy. When the epiblast was confronted with either hypoblast or germ wall, it underlapped them at the region of contact.

Animals↗
PubMed1981

Cell junctions in explanted tissues from early chick embryos.

Hypoblast and definitive endoblast derived from young chick embryos were explanted and grown for 24 h in culture. The junctional complexes which characterise these tissues were studied on freeze-fracture replicas and thin sections. Cell membranes of the hypoblast displayed tight junctions only, disposed in randomly arranged strands or narrow belts which included many discontinuous strands. The definitive endoblast showed tight and gap junctions as well as desmosomes in close association with the tight junctions. It is suggested that the differences between the two types of tissue may be related to cell cohesiveness, which appears to be relatively low in the hypoblast and high in the definitive endoblast.

Animals↗
PubMed1981

The behaviour of embryonic chick and quail tissues in culture.

Pieces of tissue were dissected from early chick and quail embryos (Stages XIII and XIV of Eyal-Giladi & Kochav, 1976; and stages 3-5 of Hamburger & Hamilton, 1951). These tissues were taken from three different regions of the early embryos, and from eight different regions of the older ones, and were derived mainly from the lower layer. Epiblast tissues were also used. The experiments were designed to test the ability of one tissue to penetrate another. A single tissue was grown in culture in a Falcon dish for 18-24 h until it had formed a coherent sheet of cells (Explant I). A second tissue was then combined with it in one of two ways: (a) A small piece of tissue (Explant II) was explanted on top of Explant I. In most cases Explant II penetrated through Explant I and spread on the Falcon dish. (b) Another small piece of tissue (Explant III) was explanted beside (in confrontation with) Explant I. Usually, Explant III penetrated into Explant I rather than vice versa. The results were analysed to see if there were any variations in behaviour of the different tissues. The main result was that important differences were found to exist between certain types of chick and quail cells when grown in culture; the implications of this finding for the widely used technique of xenoplastic grafting are mentioned. Another result was that Explant I was more likely to be penetrated when the second tissue was placed on top of it (Explant II) than when it was confronted with it (Explant III). The significance of these results is discussed.

Age Factors↗
PubMed1980

An experimental analysis of somite segmentation in the chick embryo.

In a previous paper it was suggested that collagen fibrils play an important role in the process of somite segmentation. This paper was designed mainly to test that concept. In one series of experiments, embryos were treated with either alpha, alpha'-dipyridyl or L-azetidine-2-carboxylic acid, which are analogues that interfere with the formation of normal collagen. The reagents led to a reduction in the numbers of somites that formed, as well as to the production of other anomalies such as overall diminution in size and retardation. The older the embryo at the time of treatment, the further posteriorly were the major anomalies located. It is concluded that these results lend some support to the concept. In a second series of experiments an incision was made along one side of the neural tube and notochord to separate it from the segmental plate on one side. The result was that many more somites formed on the unoperated (control) side of the embryo than on the operated side. It is concluded that these results also lend support to the concept; but that they are of interest also in relation to the mechanisms involved in the control of somite numbers. In a third group of experiments, attempts were made to obtain somites in the absence of endoderm. Although this was not possible using surgery, it was achieved by treating the young embryos with U.V. irradiation. It was concluded that the presence of endoderm is not essential for the segmentation of mesoderm.

2,2'-Dipyridyl↗
PubMed1980

Behavioural properties of chick somitic mesoderm and lateral plate when explanted in vitro.

Tissue culture, time-lapse cinematographic and electron microscopic techniques have been used to study the properties of chick mesoderm at several stages of differentiation. Lateral plate, unsegmented mesoderm (segmental plate), and newly formed somites were dissected from stage-12 embryos, whilst dermo-myotomes and sclerotomes were dissected from stage-18 embryos. Each type of mesoderm was found to exhibit a characteristic pattern of behaviour. The explants from the unsegmented mesoderm from the newly formed somites and from the older embryos could be placed in a developmental sequence; with increasing differentiation they settled and spread on the substrate more readily, whether explanted as pieces of tissue or as individual cells, and it was concluded that this implied an increased adhesion to the substrate. Similarly, with increasing differentiation, the cells segmented at a faster rate. No significant differences could be discerned in the internal structure of the different types of cells, although differences in the general shape were apparent. The lateral plate mesoderm cells, which bear some resemblances to the unsegmented mesoderm cells in the embryo, also show some morphological resemblances to them in vitro. However, the lateral plate cells had a much greater success in attaching to glass or platic substrates. They were also found to have the highest speed of locomotion of all the tissues studied, whereas the unsegmented had the lowest. It is concluded therefore, that although cells may look similar to one another morphologically, their behaviour may differ greatly, probably because they are already partially determined.

Animals↗
PubMed1979

The mechanism of somite segmentation in the chick embryo.

The segmentation of somites in the chick embryo has been studied by transmission and scanning electron microscopy (stages 8-14). The segmental plate mesoderm consists of loosely arranged mesenchymal cells, whereas the newly formed somites are composed of elongated, spindle-shaped cells arranged radially around a lumen, the myocoele. The diamter of each somite is thus two cells plus the myocoele. Two major factors appear to be responsible for the change in cell shape at segmentation: (1) Each prospective somite cell becomes anchored at one end to the adjacent epithelia (i.e. the neural tube, the notochord, the ectoderm, the endoderm or the aorta) by means of collagen fibrils. These fibrils are already present in the segmental plate before the somites begin to form. (2) A change in cell-to-cell adhesiveness causes the free ends of these cells to adhere to one another. (Bellairs, Curtis & Sanders, 1978). This adhesion is then supplemented by the development of tight junctions proximally in the somite. Because it is anchored at both ends, each somite cell is under tension in much the same way as a fibroblast cell in tissue culture is under tension. Each somite cell therefore becomes elongated and the somite as a whole accommodates its general shape to that of the space available between the adjacent tissues. The arrangement of the cells in the more differentiated somites (stages 17-18) has also been examined and it has been found that the chick resembles Xenopus in that the myotome cells undergo rotation and become orientated in an anteroposterior direction.

Animals↗
PubMed1978

Effect of fixation on cell membrane of early embryonic material as observed on freeze-fracture replicas.

Some cell membranes of chemically fixed, cryoprotected and freeze-fractured chick embryonic material are characterized by 150-250 nm diameter circular smooth blebs. No such blebs are observed, however, on cryoprotected, rapidly frozen but not chemically fixed samples. It is concluded, therefore, that fixation with buffered glutaraldehyde and not cryoprotection with glycerol is mainly responsible for the appearance of the blebs. The suggestion that blebs represent a budding virus is not tenable, since blebs are present in material obtained from virus free embryos.

Animals↗
PubMed1978

Cleavage in the chick embryo.

Chick embryos ranging from the stage of first cleavage to that of about 700 cells were removed from the oviduct and examined by transmission electron microscopy. Beneath the cell membrane is yolk-free cortical region containing microfilaments. Beneath this lies cytoplasm which contains yolk spheres which are graded in size, the dorsal ones being smaller than the ventral ones. The subgerminal periblast possesses a greater proportion of yolk to cytoplasm than do the cells proper, but it merges with the cytoplasm at the incomplete borders of the 'open' cells. Specialized accumulations of membranes lie in the marginal periblast, and it is suggested that they play a role in cell membrane formation.

Animals↗