An analysis of the aggregation and morphogenesis of area opaca endoderm cells from the primitive-streak chick embryo

Development ◽  
1979 ◽  
Vol 51 (1) ◽  
pp. 121-135
Author(s):  
Nadine Milos ◽  
Sara E. Zalik ◽  
Robert Phillips
Keyword(s):  
Cell Adhesion ◽  
Chick Embryo ◽  
Intrinsic Property ◽  
Primitive Streak ◽  
Chick Embryos ◽  
Embryonic Cells ◽  
Thin Walls ◽  
Cell Layers ◽  
Aggregative Behaviour

The aggregative behaviour and subsequent morphogenesis of extra-embryonic endoderm cells from primitive-streak chick embryos have been investigated. A relatively pure population of area opaca endoderm cells was obtained by differential dissociation, which involves partial separation of epiblast and endoderm cell clumps by sieving through Nitex mesh. For aggregation studies cells were cultured in rotating flasks in Leibovitz (L-15) medium, in saline or in saline supplemented with glucose (1 mg/ml). Aggregation was monitored using the Coulter Counter. In these three media aggregation is rapid; by 10 min an average of 61% of the population had aggregated, to reach a plateau at 30 min when an average percent adhesion value of 83 % was obtained. The aggregates in L-15 medium were large and compact. After several days in culture, they cavitated and formed smooth hollow vesicles with thin walls composed of one or a few cell layers. Aggregates formed in PCS were smaller and looser in appearance; the addition of glucose resulted in a certain degree of compaction. Some morphogenesis occurred under these conditions with the aggregates developing numerous irregular cavities. These experiments suggest that some of the factors that affect cell adhesion in early embryonic cells can be studied in vitro. The results also indicate that the ability to cavitate is an intrinsic property of the endoderm cells of the area opaca since this occurs in the absence of epiblast or mesoderm.

scholarly journals Effect of cortisol acetate on collagen biosynthesis and on the activities of prolyl hydroxylase, lysyl hydroxylase, collagen galactosyltransferase and collagen glucosyltransferase in chick-embryo tendon cells

1977 ◽  
Vol 164 (3) ◽  
pp. 533-539 ◽  
Author(s):  
A Oikarinen
Keyword(s):  
Chick Embryo ◽  
Enzyme Activities ◽  
Collagen Synthesis ◽  
Enzyme Synthesis ◽  
Prolyl Hydroxylase ◽  
Chick Embryos ◽  
Isolated Cells ◽  
Lysyl Hydroxylase ◽  
Tendon Cells

Collagen synthesis and the activities of prolyl hydroxylase, lysyl hydroxylase, collagen galactosyltransferase and collagen glucosyltransferase were studied in isolated chick-embryo tendon cells after the administration of cortisol acetate to the chick embryos. When the steroid was injected 1 day before isolation of the tendon cells, collagen synthesis was decreased, even though the enzyme activities were not changed. When cortisol acetate was given as repeated injections over a period of 4 days, both collagen synthesis and the enzyme activities decreased. The hydroxylase activities decreased even more than the two collagen glycosyltransferase activities, both in isolated cells and in whole chick embryos. The amount of prolyl hydroxylase protein diminished to the same extent as the enzyme activity, indicating that cortisol acetate inhibits enzyme synthesis. The inhibitory effect of cortisol acetate on collagen synthesis and on the enzyme activities was partially reversible in 3 days. Total protein synthesis was completely restored within this time. Only massive doses of cortisol acetate inhibited collagen synthesis in vitro. Additional experiments indicated that cortisol acetate did not decrease the rate of the enzyme reactions when added directly to the enzyme incubation mixtures. The results suggest that cortisol acetate decreases collagen synthesis both by its direct effect on collagen polypeptide-chain synthesis and by decreasing the activities of enzymes involved in post-translational modifications.

The Effect of Chloroacetophenone on the Inducing Capacity of Hensen's Node

Development ◽  
1962 ◽  
Vol 10 (3) ◽  
pp. 383-388
Author(s):  
M. S. Lakshmi
Keyword(s):  
Organizer Region ◽  
High Capacity ◽  
Primitive Streak ◽  
Chick Embryos

In a previous paper (Lakshmi, 1962) the effects of ω-chloroacetophenone (CAP), which is an irreversible —SH inhibitor, on the morphogenesis of chick embryos cultured in vitro were reported. Brachet (1950) suggested that the —SH-containing proteins might be active in induction. Rapkine & Brachet (1951) studied the effect of monoiodoacetate on the amphibian organizer and observed that the organizer region retained a high capacity for induction despite treatment with the inhibitor. The action of monoiodoacetate is reversible, hence it was felt desirable to investigate the action of CAP on the living organiser of chick, namely Hensen's node. Chick embryos at the primitive-streak stage were explanted in vitro by New's (1955) technique. These were treated with 0·0005 M CAP for 15 and 30 minutes, 0·001 M CAP for 15 minutes, and 0·0015 M CAP for 15 minutes. 0·1 ml. of the solution was added to the endodermal surface of the explanted embryos.

The Effect of Chloroacetophenone on Chick Embryos Cultured in vitro

Development ◽  
1962 ◽  
Vol 10 (3) ◽  
pp. 373-382
Author(s):  
M. S. Lakshmi
Keyword(s):  
Xenopus Laevis ◽  
Average Length ◽  
Primitive Streak ◽  
Poultry Farm ◽  
Chick Embryos ◽  
Normal Process ◽  
Strong Emphasis ◽  
The Brain

Brachet's (1950) strong emphasis on the role of —SH-containing proteins in the process of induction has stimulated a study of the interference in the normal process of morphogenesis of chick embryos by chloroacetophenone, which has been described by Beatty (1951) as a specific and irreversible —SH inhibitor. He studied the effect of chloroacetophenone on the development of embryos of Rana and Triturus employing different concentrations. Deuchar (1957) also studied the action of the same chemical on the embryos of Xenopus laevis and has recorded abnormalities mainly in the brain and the eye. In the present work ω-chloroacetophenone (CAP) commercially known as phenacyl chloride (ω—C6H5.CO.CH2Cl) was employed. The sample used was a B.D.H. product. Fresh fertilized hens' eggs brought from a local poultry farm were incubated at 37·5° C. for 16 to 18 hours to obtain definitive primitive-streak stages (range of length from 1·75 mm. to 2 mm.) or for about 22 hours to obtain head-process stages (average length of the head process alone 0·56 mm.).

Interactions between mesoderm cells and the extracellular matrix following gastrulation in the chick embryo

1991 ◽  
Vol 99 (2) ◽  
pp. 431-441
Author(s):  
A.J. Brown ◽  
E.J. Sanders
Keyword(s):  
Extracellular Matrix ◽  
Chick Embryo ◽  
Basement Membrane ◽  
Primitive Streak ◽  
Cell Binding ◽  
Fab Fragment ◽  
Fibronectin Receptor ◽  
In Vitro Methods

In the gastrulating chick embryo, the mesoderm cells arise from the epiblast layer by ingression through the linear accumulation of cells called the primitive streak. The mesoderm cells emerge from the streak with a fibroblastic morphology and proceed to move away from the mid-line of the embryo using, as a substratum, the basement membrane of the overlying epiblast and the extracellular matrix. We have investigated the roles of fibronectin and laminin as putative substrata for mesoderm cells using complementary in vivo and in vitro methods. We have microinjected agents into the tissue space adjacent to the primitive streak of living embryos and, after further incubation, we have examined the embryos for perturbation of the mesoderm tissue. These agents were: cell-binding regions from fibronectin (RGDS) and laminin (YIGSR), antibodies to these glycoproteins, and a Fab' fragment of the antibody to fibronectin. We find that RGDS, antibody to fibronectin, and the Fab' fragment cause a decrease in the number of mesoderm cells spread on the basement membrane, and a perturbation of cell shape suggesting locomotory impairment. No such influence was seen with YIGSR or antibodies to laminin. These results were extended using in vitro methods in which mesoderm cells were cultured in fibronectin-free medium on fibronectin or laminin in the presence of various agents. These agents were: RGDS; YIGSR; antibodies to fibronectin, fibronectin receptor, laminin and vitronectin; and a Fab' fragment of the fibronectin antiserum. We find that cell attachment and spreading on fibronectin is impaired by RGDS, antiserum to fibronectin, the Fab' fragment of fibronectin antiserum, and antiserum to fibronectin receptor. The results suggest that although the RGDS site in fibronectin is important, it is probably not the only fibronectin cell-binding site involved in mediating the behaviour of the mesoderm cells. Cells growing on laminin were perturbed by YIGSR, RGDS and antibodies to laminin, suggesting that mesoderm cells are able to recognise at least two sites in the laminin molecule. We conclude that the in vivo dependence of mesoderm cells on fibronectin is confirmed, but that although these cells have the ability to recognise sites in laminin as mediators of attachment and spreading, the in vivo role of this molecule in mesoderm morphogenesis is not yet certain.

Impaired energy metabolism as an initial step in the mechanism for 6-aminonicotinamide-induced limb malformation

Development ◽  
1980 ◽  
Vol 59 (1) ◽  
pp. 217-222
Author(s):  
Yal C. Sheffield ◽  
Robert E. Seegmiller
Keyword(s):  
Amino Acid ◽  
Energy Metabolism ◽  
Chick Embryo ◽  
Chondroitin Sulfate ◽  
Atp Synthesis ◽  
Initial Step ◽  
Chick Embryos ◽  
Impaired Energy Metabolism ◽  
All Treatment

The analogue and antagonist of nicotinamide, 6-aminonicotinamide (6-AN), impairs cartilage formation and results in shortening of the limbs when administered to chick embryos. Studies have shown that 6-AN forms an abnormal NAD analogue which inhibits the activity of NAD-dependent enzymes associated with production of ATP. To determine if an effect on ATP synthesis might be associated with the mechanism of teratogenesis in the chick embryo, ATP levels of cartilage from day-8 chick embryos treated in vitro were assayed in relation to biosynthesis of protein, DNA and chondroitin sulfate. Incorporation of 35SO4− was inhibited by 6 h of treatment with 10 µg/ml of 6-AN, whereas incorporation of [3H]thymidine and [3H]amino acid was not inhibited until 12 h. Incorporation of [3H]- glucosamine was increased at all treatment times. A decrease in the level of ATP preceded any detectable inhibition of precursor incorporation. These results are consistent with the hypothesis that 6-AN inhibits chondroitin sulfate synthesis through a reduction in the level of ATP in chondrocytes.

In Vitro Evaluation of Plasticizer Activity on the Growth and Metabolism of Chick Embryo Fibroblasts and on the Development of Chick Embryo Lungs

1992 ◽  
Vol 20 (6) ◽  
pp. 475-482 ◽  
Author(s):  
G Stabellini ◽  
O Fiocchi ◽  
A Pellati ◽  
A Caruso
Keyword(s):  
Cell Proliferation ◽  
Protein Synthesis ◽  
Chick Embryo ◽  
Primary Cultures ◽  
Embryonic Cells ◽  
Cytostatic Effect ◽  
Dna And Protein Synthesis ◽  
Embryo Fibroblasts ◽  
Ethylhexyl Phthalate

Administration of di(2-ethylhexyl) phthalate (DEHP) to primary cultures of chick embryo fibroblasts brought about a decrease in cell proliferation rate after 48 h and an inhibition of both DNA and protein synthesis measured by [3H]thymidine and [3H]leucine, respectively, after 48h. The growth of chick embryo lung rudiments in vitro was also depressed by DEHP treatment. Lung rudiment were smaller in DEHP-treated embryos after 6 days' treatment. These results indicate that DEHP has a cytostatic effect on embryonic cells and tissues. L'administration de di(2-éthylhexyl) phthalate (DEHP) à des cultures primaires de fibroblastes d'embryon de poulet a provoqué une diminution du taux de prolifération cellulaire après 48 heures et une inhibition de la synthèse à la fois d'ADN et de protéines mesurée, respectivement, par [3H]thymidine et [3H]leucine après 48 h. La croissance des ébauches pulmonaires de l'embryon de poulet in vitro a également été abaissée par le traitement DEHP. Les ébauches pulmonaires étaient plus petites dans les embryons traités au DEHP après un traitement de 6 jours. Ces résultats indiquent que le DEHP a un effet cytostatique sur les cellules et tissus embryonnaires.

scholarly journals THE DEPENDENCE OF HEAD CURVATURE ON THE DEVELOPMENT OF THE HEART IN THE CHICK EMBRYO

1937 ◽  
Vol 14 (2) ◽  
pp. 229-231 ◽  
Author(s):  
C. H. WADDINGTON
Keyword(s):  
Chick Embryo ◽  
Blood Vessels ◽  
Brain Region ◽  
Normal Development ◽  
Anterior Part ◽  
Blood System ◽  
Chick Embryos ◽  
Aortic Arches ◽  
The Brain

1. The heart was removed from chick embryos of seven to twelve somites, and the embryos cultivated in vitro. The operation abolished the normal twisting of the anterior part of the embryo on to its left side and the general bending of the brain region into an arc. These two processes therefore seem to be dependent on the normal development of the heart. 2. The embryos showed a bending of the forebrain relative to the midbrain, which is therefore independent of the development of the heart. 3. The embryonic blood system, including the aortic arches, developed normally in many cases, but the blood vessels became enormously dilated. 4. The lateral evaginations of the foregut and the visceral arch mesenchyme underwent the first stages of differentiation in atypical positions, seemingly independently of each other or of any other structures.

The Factor Affecting the Spreading of Chondrocytes upon Inorganic Substrate

1973 ◽  
Vol 13 (1) ◽  
pp. 193-204
Author(s):  
M. TAKEICHI
Keyword(s):  
Conditioned Medium ◽  
Calf Serum ◽  
Fresh Medium ◽  
Foetal Calf Serum ◽  
Chick Embryos ◽  
Embryonic Cells ◽  
Mass Cultures ◽  
Rounded Form ◽  
Inorganic Substrate

The effect of conditioned medium (CM) prepared from mass-cultures of chick embryonic cells was studied on the spreading behaviour of chondrocyte derived from sterna of 16-day-old chick embryos. Freshly dissociated chondrocytes exhibited a quite rounded form, and this shape did not change when they were cultured with fresh medium (Eagle's MEM + 6% foetal calf serum) in vitro for several days. Non-dialysable material(s) in CM added into the fresh medium stimulated the formation of pseudopods of chondrocytes, without primarily affecting the synthesis of chondroitin sulphates. Such an activity of CM was not lost after boiling, but it was lost following treatment with proteases. The chondrocytes covered with newly deposited acidmucopoly-saccharides were insensitive to the effect of CM, but they became sensitive to form pseudopods after treatment of the cells with chondroitinase. These results suggest that CM contains a macromolecular material(s) to enhance the motility or adhesiveness of chondrocytes.

Simvastatin Reduces the in Vitro Adhesion of Sickle Cell Disease Neutrophils to Endothelial Layers

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2489-2489 ◽  
Author(s):  
Andreia A Canalli ◽  
Renata P. Ferreira ◽  
Sara T.O. Saad ◽  
Nicola Conran ◽  
Fernando F. Costa
Keyword(s):  
Cell Adhesion ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Leukocyte Adhesion ◽  
Cell Disease ◽  
Tnf Α ◽  
Cell Layers ◽  
Anti Inflammatory ◽  
Vitro Effect

Abstract Leukocytes may have a propagating and, possibly, initiating role in sickle cell disease (SCD) vaso-occlusion. Endothelial dysfunction contributes to the vaso-occlusion process and leads to inflammation, leukocyte and red cell adhesion. Markers of neutrophil activation are also increased in SCD, in association with increased levels of circulating cytokines and increased leukocyte adhesion. In animal models, vaso-occlusion causes hypoxia/reperfusion, leading to vascular endothelium damage and an inflammatory response. We postulate that anti-inflammatory agents may reduce the participation of activated endothelium in the vaso-occlusive process. Statins are commonly used to treat arteriosclerosis and have anti-inflammatory effects that include a regulatory action on endothelial function, reduced oxidative stress and inflammation. The objective of this study was to investigate the in vitro effect of simvastatin on the adhesion of sickle neutrophils to activated endothelial cell layers (HUVEC). Neutrophils (Neu) were isolated from the peripheral blood of healthy controls (ConNeu) and SCD (SCDNeu) individuals in steady state over ficoll-paque gradients. Cell adhesion (2×106 cell/ml in Ham’s F12 K) to cultured human umbilical vein endothelial cells (HUVEC) grown to confluence was assessed using static adhesion assays. HUVEC cells were treated with or without 1 μg/ml simvastatin for 6 hours in the absence or presence of a 10nM TNF-α activating stimulus (3 hours) before allowing adhesion of Neu to the cell layers (30 min, 37°C, 5%CO2). Neu from SCD patients demonstrated a significantly greater adhesion to HUVEC than ConNeu (20.5 ± 1.9% compared to 13.8 ± 1.7 %; n=15; p<0.02; Mann Whitney test). Subsequently, Neu from patients and controls were allowed to adhere to endothelial layers previously treated with simvastatin; adhesion was not significantly different to the adhesion of Neu to nonsimvastatin treated HUVEC (16.7 ± 3.2% for ConNeu; n=8, p>0.05 and 19.8 ±2.7% for SCDNeu; n=11, p>0.05, paired t test). Pre-treatment of HUVEC with the cytokine TNF-α increased the adhesion of SCD and Con Neu to HUVEC (40.9 ± 5.4%; 28.9 ± 5.0%, respect, N>8, P<0.01 compared to adhesion to non-activated HUVEC). Interestingly, when the endothelium layer was protected with simvastatin and then stimulated with TNF-α, SCDNeu adhesion was significantly diminished (reduced to 31.3% ± 3.6%; n=11, p<0.005 comp. to adhesion to non-simvastatin-treated HUVEC); in contrast, no difference in the adhesion of ConNeu to HUVEC treated with TNF-α and simvastatin was observed (31.9 ± 5.8%, n=8, p>0.05 for ConNeu). In conclusion, data indicate that under in vitro inflammatory conditions, simvastatin appears to protect endothelium layers and reduces SCD leukocyte adhesion. We speculate that statins may have anti-inflammatory properties and, as such, may be useful for diminishing endothelial activation and, in turn, preventing the adhesion of leukocytes adhesion to the vascular wall in SCD, a mechanism that is essential to the vaso-occlusive process.

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