Ultrastructural immunocytochemical localization of fibronectin in the early chick embryo

Development ◽  
1982 ◽  
Vol 71 (1) ◽  
pp. 155-170
Author(s):  
E. J. Sanders
Keyword(s):  
Chick Embryo ◽  
Basal Lamina ◽  
Regional Differences ◽  
Developmental Stages ◽  
Primitive Streak ◽  
Extracellular Material ◽  
Cell Surfaces ◽  
Mesoderm Cell ◽  
Stage Of Development ◽  
Primary Location

Fibronectin was localized using peroxidase and biotin-avidin-ferritin techniques in developmental stages of the chick embryo from laying to primitive streak formation. The primary location of fibronectin at all stages examined was the basal lamina of the epiblast and its associated extracellular material. The remaining tissues showed little or no immunochemical deposit. At the primitive streak stage of development there were some regional differences in the density of the deposit on the basal lamina. Closely adjacent to the primitive streak, where the basal lamina is fragmentary, deposit was sparse or absent. In this, and other regions, mesoderm cell surfaces did not stain except where they closely approached the stained basal lamina or interstitial bodies. Staining was variable in the basal lamina anterior to the primitive streak, but in a number of cases particularly heavy deposit was noted overlying the crescent of late hypoblast. This area seemed to correspond with the anterior fibronectin- rich band reported by others using immunofluorescence localization.

Labelling of basement membrane constituents in the living chick embryo during gastrulation

Development ◽  
1984 ◽  
Vol 79 (1) ◽  
pp. 113-123
Author(s):  
Esmond J. Sanders
Keyword(s):  
Chick Embryo ◽  
Basement Membrane ◽  
Concanavalin A ◽  
Primitive Streak ◽  
Basal Surface ◽  
Stage Of Development ◽  
Pass Through ◽  
The Relationship

The basement membrane of the living chick embryo epiblast has been labelled with ultrastructural markers in order to study the movement and turnover of this structure during gastrulation. Two problems were addressed in these experiments. Firstly, to what extent does the basement membrane move medially with the epiblast during morphogenesis? Secondly, what is the relationship to the basement membrane of the so-called interstitial bodies? The ultrastructural markers used were concanavalin A conjugated to ferritin and fibronectin antibodies conjugated to peroxidase. Embryos were cultured using the technique of New, and the label was applied to the periphery of the basal surface of the epiblast through a hole in the endoblast at the early primitive streak stage of development. The embryos were then allowed to develop to the full primitive streak stage in the presence of the label. When the position of the label was determined after incubation, it was found to have accumulated in large amounts at the edge of the primitive streak at the point where the basement membrane is disrupted. This indicates that constituents of the basement membrane are transported medially with the epiblast cells and are sloughed off as the latter pass through the primitive streak. This movement of basement membrane constituents is counter to the direction of migration of the underlying mesoderm cells. When embryos are exposed to label for only 1 h, then washed and incubated for a further three hours, the marker was found in the interstitial bodies and not distributed throughout the basement membrane itself. This suggests that the interstitial bodies, which have been implicated in influencing the migration of the mesoderm cells, are turnover products of the basement membrane to which they are attached.

scholarly journals AUTORADIOGRAPHIC STUDIES OF THE UTILIZATION OF S35-SULFATE BY THE CHICK EMBRYO

1957 ◽  
Vol 3 (2) ◽  
pp. 231-238 ◽  
Author(s):  
Perry M. Johnston ◽  
Cyril L. Comar
Keyword(s):  
Chick Embryo ◽  
Basement Membrane ◽  
Chondroitin Sulfate ◽  
Developmental Stages ◽  
Primitive Streak ◽  
Intracellular Accumulation ◽  
Histological Differentiation ◽  
Membrane Area ◽  
Thin Ring ◽  
Inorganic Sulfate

From studies of autoradiograms of various developmental stages of the chick embryo containing S35 given us sulfate it was determined that as early as Stages 3+ and 4 there is a selective utilization or accumulation of sulfate by the various parts. The earliest accumulation site is the axial portion of the primitive streak and the floor of the groove. Later S35 was found in the head process, Hensen's node, notochord, amniocardiac vesicle, wall of the omphalomesenteric vein, endocardium, subendocardial jelly, mesenchyme destined to become cartilage, basement membrane area of the gut, and a mucopolysaccharide layer formed on the free surface of the stomach. The early notochordal localizations of S35 coincide with the region in which a thin ring of chondroitin sulfate is subsequently laid down. However, it is apparent that there is an intracellular accumulation of inorganic sulfate by the chondroitin-forming cells prior to the time they produce sufficient chondroitin sulfate to be demonstrable histochemically. It was interesting to note that the endocardium appears to concentrate sulfate that later apparently finds its way into the subendocardial jelly. The fact that those mesenchymal cells which later form chondroblasts begin to utilize sulfate selectively before histological differentiation is apparent was determined. In addition, the presence of sulfate-containing substances in the forming basement membrane of the gut would seem to indicate that sulfate is important in the histological differentiation of this membrane.

Identification of a novel growth factor with transforming activity secreted by individual chick embryos

Development ◽  
1990 ◽  
Vol 109 (4) ◽  
pp. 905-910 ◽  
Author(s):  
J. Smith ◽  
J.C. McLachlan
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Chick Embryo ◽  
Primitive Streak ◽  
Soft Agar ◽  
Heat Stable ◽  
Stage Of Development ◽  
Transforming Activity ◽  
Embryo Tissue ◽  
Differential Responsiveness

Previously we have developed a microassay for anchorage independent growth (AIG) of fibroblasts in soft agar, which can detect very small quantities of transforming growth factors (TGFs). Using this assay, we have shown that small pieces of dissected chick embryo tissue will stimulate AIG of both NR6 and NRK 49f cells, and that this property can be used to map production of growth factors with transforming activity in individual early embryos. We now show that this activity can be transferred to conditioned medium (CM) prepared using chick embryo tissue. Using two cell lines with differential responsiveness to TGFs, and by coincubating normal and heat-treated CM with trypsin, Con-A and neutralising antibodies, we show that CM contains at least two different growth factors with transforming activity. One of these is heat-stable, and stimulates colony formation in NRK 49f cells in the presence of EGF, but not in its absence. This activity corresponds to a TGF beta-like molecule. The other component is a heat-labile glycoprotein, which has TGF alpha-like properties, but does not appear to behave like known TGFs with these properties. It therefore appears to be a novel growth factor. Both activities are present from the intermediate primitive streak stage of development.

scholarly journals Crayfish hemocytes develop along the granular cell lineage

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Fang Li ◽  
Zaichao Zheng ◽  
Hongyu Li ◽  
Rongrong Fu ◽  
Limei Xu ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cell Lineage ◽  
Granular Cell ◽  
Marker Genes ◽  
Hematopoietic Tissue ◽  
Differentiated Cells ◽  
Stage Of Development ◽  
Almost All ◽  
Relationship Of

AbstractDespite the central role of hemocytes in crustacean immunity, the process of hemocyte differentiation and maturation remains unclear. In some decapods, it has been proposed that the two main types of hemocytes, granular cells (GCs) and semigranular cells (SGCs), differentiate along separate lineages. However, our current findings challenge this model. By tracking newly produced hemocytes and transplanted cells, we demonstrate that almost all the circulating hemocytes of crayfish belong to the GC lineage. SGCs and GCs may represent hemocytes of different developmental stages rather than two types of fully differentiated cells. Hemocyte precursors produced by progenitor cells differentiate in the hematopoietic tissue (HPT) for 3 ~ 4 days. Immature hemocytes are released from HPT in the form of SGCs and take 1 ~ 3 months to mature in the circulation. GCs represent the terminal stage of development. They can survive for as long as 2 months. The changes in the expression pattern of marker genes during GC differentiation support our conclusions. Further analysis of hemocyte phagocytosis indicates the existence of functionally different subpopulations. These findings may reshape our understanding of crustacean hematopoiesis and may lead to reconsideration of the roles and relationship of circulating hemocytes.

Cell proliferation in the gastrulating chick embryo: a study using BrdU incorporation and PCNA localization

Development ◽  
1993 ◽  
Vol 118 (2) ◽  
pp. 389-399 ◽  
Author(s):  
E.J. Sanders ◽  
M. Varedi ◽  
A.S. French
Keyword(s):  
Cell Proliferation ◽  
Chick Embryo ◽  
Cell Density ◽  
Generation Time ◽  
Primitive Streak ◽  
S Phase ◽  
Nuclear Antigen ◽  
Brdu Incorporation ◽  
Similar Proportion ◽  
Differential Growth

Cell proliferation in the gastrulating chick embryo was assessed using two independent techniques which mark cells in S phase of the mitotic cycle: nuclear incorporation of bromodeoxyuridine (BrdU) detected immunocytochemically and immunolocalization of proliferating cell nuclear antigen (PCNA). Computer-reconstructed maps were produced showing the distribution of labelled nuclei in the primitive streak and the cell layers. These distributions were also normalized to take into account regional differences in cell density across the embryo. Results from a 2 hour pulse of BrdU indicated that although cells at caudal levels of the primitive streak showed the highest incorporation, this region showed a similar proportion of labelled cells to the surrounding caudal regions of the epiblast and mesoderm when normalized for cell density. The entire caudal third of the embryo showed the highest proportion of cells in S phase. Cells of Hensen's node showed a relatively low rate of incorporation and, although the chordamesoderm cells showed many labelled nuclei, this appeared to be a reflection of a high cell density in this region. Combining this result with results from a 4 hour pulse of BrdU permitted mapping of cell generation time across the entire embryo. Generation times ranged from a low value of approximately 2 hours at caudal levels of both the epiblast and mesoderm, to an upper value of approximately 10 hours in the rostral regions of the primitive streak, in the mid-lateral levels of the epiblast and in the chordamesoderm rostral to Hensen's node. Cells at caudal regions of the primitive streak showed a generation time of approximately 5 hours. Taking into account that cells are generally considered to be continuously moving through the primitive streak, we conclude that cell division, as judged by generation time, is greatly reduced during transit through this region, despite the presence there of cells in S phase and M phase. Immunocytochemical localization of PCNA-positive nuclei gave generally similar distributions to those obtained with BrdU incorporation, confirming that this endogenous molecule is a useful S-phase marker during early embryogenesis. Mid-levels and caudal levels of the primitive streak showed the highest numbers of positive nuclei, and the highest proportion of labelling after cell density was accounted for. As with BrdU incorporation, the highest proportions of PCNA-positive nuclei were found towards the caudal regions of the epiblast and mesoderm. These results suggest that the differential growth of the caudal region of the embryo at this time is a direct consequence of elevated levels of cell proliferation in this region.(ABSTRACT TRUNCATED AT 400 WORDS)

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