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

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.

Development ◽  
1962 ◽  
Vol 10 (3) ◽  
pp. 373-382
Author(s):  
M. S. Lakshmi

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.).


Development ◽  
1979 ◽  
Vol 51 (1) ◽  
pp. 121-135
Author(s):  
Nadine Milos ◽  
Sara E. Zalik ◽  
Robert Phillips

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.


Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 127-140
Author(s):  
Kazuko Uno

Seven different embryoid body (EB) lines of mouse teratocarcinoma were isolated from a single EB. With regard to each of the lines, a comparison was made of the following developmental properties, including potentiality: (1) cavity formation in a short term intraperitoneal passage, (2) growth in vivo, (3) cardiac muscle differentiation in vitro following intraperitoneal passage and (4) differentiation of solid tumours in vivo. These lines could be divided into three distinct groups withrespect to their capacity for cardiac muscle differentiation. It has been shown that a high capacity for celldifferentiation in vitro correlates well with the capacity for cavity formation of an EB during the in vivo period. This cavity formation was followed by the appearance of primitive-streak-like structures, from which mesodermal cells were subsequently formed.


In a series of tissue cultures of fragments taken from different parts of young chick embryos, at various stages, it was found that differentiation frequently occurred, the characteristic cells of highly specialised tissues appearing in cultures in which they could not possibly have been present at explantation. The most striking differentiations were the development of red blood corpuscles, capillary vessels, nerve cells with axons and of rhythmically contracting cardiac muscle. This paper describes a simple method by which may be obtained regularly and quickly the development of very large numbers of red cells, and an account is given of the histology of the cultures. It is intended to be introductory to a physiological study, now in progress, of the conditions of hæmatopoiesis Literature. Erythropoiesis in vitro has been reported by several authors, but it has not been thoroughly investigated and, with the exceptions of the works of Slonimski (1930, a , 1931) and Shipley (1915-16), the earlier papers have con­cerned the somewhat sporadic appearance of small numbers of erythrocytes. In the earlier works, also, the explants were derived from hæmatopoietic organs, or from that part of the embryo in which blood would normally have developed at latest quite soon after the time at which the experiment was made. The present paper, on the other hand, is based principally upon cultures of fragments of the primitive streak—that is, upon explants of presumptively hæmatopoietic cells isolated before they had arrived at the normal hæmatopoietic region of the embryo. The literature may be very briefly summarised as follows : Shipley (1915-16) made plasma cultures from the area opaca of chick embryos at a time prior to the formation of the blood islands, and obtained the differen­tiation of erythrocytes from amœboid cells. Erythropoiesis is reported by N. G. and A. L. Ghlopin (1925) in cultures of Axolotl spleen; by Erdmann, Eisner, and Laser (1925-26), in cultures of embryonal rat spleen; by Freifeld and Ginsburg (1927) in cultures of rabbit adrenals; by de Haan (1928-29) in cultures of blood cells of the horse; by Timofejewsky and Benewolenskaja (1929) in cultures of blood from a case of acute myeloid leukaemia, and by Benewolenskaja (1930) in cultures of embryonal human liver. Slonimski (1930 a , 1931), using Rana fusca and Axolotl embryos, excised the blood island zone at early stages, and kept it as a culture enclosed in a sheath of epiblastic epithelium. The little cyst became full of red blood in an abundant plasma, and there were vessels with endothelial walls.


1913 ◽  
Vol 17 (2) ◽  
pp. 182-191 ◽  
Author(s):  
Ragnvald Ingebrigtsen

1. The brains of chick embryos, of cats six weeks old, of rabbits two months old, and of dogs three weeks old, when cultivated in vitro, develop long filaments which, according to their growth and their anatomical and tinctorial characters, must be considered as true axis cylinders. 2. Similar structures develop from spinal ganglia of rabbits seven months old, and from the spinal cord of cats six weeks old, and of rabbits two months old. 3. When severed from their origin by section these threads undergo degenerative changes which do not appear after nine hours, but which are seen after twenty hours, and continue until in the course of the following two days the thread degenerates completely. 4. After twenty hours the development of new axis cylinders from the central part of the cut fibers is observed.


Reports ◽  
2021 ◽  
Vol 4 (1) ◽  
pp. 7
Author(s):  
Lea Mallo ◽  
Valentin Do Sacramento ◽  
Christian Gachet ◽  
Susan Chan ◽  
Philippe Kastner ◽  
...  

In vitro, the differentiation of megakaryocytes (MKs) is improved by aryl-hydrocarbon receptor (AHR) antagonists such as StemRegenin 1 (SR1), an effect physiologically recapitulated by the presence of stromal mesenchymal cells (MSC). This inhibition promotes the amplification of a CD34+CD41low population able to mature as MKs with a high capacity for platelet production. In this short report, we showed that the emergence of the thrombocytogenic precursors and the enhancement of platelet production triggered by SR1 involved IKAROS. The downregulation/inhibition of IKAROS (shRNA or lenalidomide) significantly reduced the emergence of SR1-induced thrombocytogenic population, suggesting a crosstalk between AHR and IKAROS. Interestingly, using a proximity ligation assay, we could demonstrate a physical interaction between AHR and IKAROS. This interaction was also observed in the megakaryocytic cells differentiated in the presence of MSCs. In conclusion, our study revealed a previously unknown AHR/ IKAROS -dependent pathway which prompted the expansion of the thrombocytogenic precursors. This AHR- IKAROS dependent checkpoint controlling MK maturation opens new perspectives to platelet production engineering.


Development ◽  
1992 ◽  
Vol 116 (Supplement) ◽  
pp. 157-165 ◽  
Author(s):  
R. S. P. Beddington ◽  
P. Rashbass ◽  
V. Wilson

Mouse embryos that are homozygous for the Brachyury (T) deletion die at mid-gestation. They have prominent defects in the notochord, the allantois and the primitive streak. Expression of the T gene commences at the onset of gastrulation and is restricted to the primitive streak, mesoderm emerging from the streak, the head process and the notochord. Genetic evidence has suggested that there may be an increasing demand for T gene function along the rostrocaudal axis. Experiments reported here indicate that this may not be the case. Instead, the gradient in severity of the T defect may be caused by defective mesoderm cell movements, which result in a progressive accumulation of mesoderm cells near the primitive streak. Embryonic stem (ES) cells which are homozygous for the T deletion have been isolated and their differentiation in vitro and in vivo compared with that of heterozygous and wild-type ES cell lines. In +/+ ↔ T/T ES cell chimeras the Brachyury phenotype is not rescued by the presence of wild-type cells and high level chimeras show most of the features characteristic of intact T/T mutants. A few offspring from blastocysts injected with T/T ES cells have been born, several of which had greatly reduced or abnormal tails. However, little or no ES cell contribution was detectable in these animals, either as coat colour pigmentation or by isozyme analysis. Inspection of potential +/+ ↔ T/T ES cell chimeras on the 11th or 12th day of gestation, stages later than that at which intact T/T mutants die, revealed the presence of chimeras with caudal defects. These chimeras displayed a gradient of ES cell colonisation along the rostrocaudal axis with increased colonisation of caudal regions. In addition, the extent of chimerism in ectodermal tissues (which do not invaginate during gastrulation) tended to be higher than that in mesodermal tissues (which are derived from cells invaginating through the primitive streak). These results suggest that nascent mesoderm cells lacking the T gene are compromised in their ability to move away from the primitive streak. This indicates that one function of the T genemay be to regulate cell adhesion or cell motility properties in mesoderm cells. Wild-type cells in +/+ ↔ T/T chimeras appear to move normally to populate trunk and head mesoderm, suggesting that the reduced motility in T/T cells is a cell autonomous defect


Development ◽  
1997 ◽  
Vol 124 (21) ◽  
pp. 4243-4252 ◽  
Author(s):  
S.K. Kim ◽  
M. Hebrok ◽  
D.A. Melton

The role of the notochord in inducing and patterning adjacent neural and mesodermal tissues is well established. We provide evidence that the notochord is also required for one of the earliest known steps in the development of the pancreas, an endodermally derived organ. At a developmental stage in chick embryos when the notochord touches the endoderm, removal of notochord eliminates subsequent expression of several markers of dorsal pancreas bud development, including insulin, glucagon and carboxypeptidase A. Pancreatic gene expression can be initiated and maintained in prepancreatic chick endoderm grown in vitro with notochord. Non-pancreatic endoderm, however, does not express pancreatic genes when recombined with the same notochord. The results suggest that the notochord provides a permissive signal to endoderm to specify pancreatic fate in a stepwise manner.


Development ◽  
1964 ◽  
Vol 12 (4) ◽  
pp. 609-619
Author(s):  
Anna Hell

Enormous progress has been made in the last few years towards the elucidation of the mechanism of protein synthesis, and great interest is centred on the steps leading to cellular differentiation and specific protein synthesis. We know that genetic information is passed on from one generation of cells to the next by deoxyribonucleic acid (DNA), and that this material directs all protein synthesis by the intermediary of the different types of ribonucleic acid (RNA). A simple in vitro system described by O'Brien (1959) seemed to offer an excellent tool for the study of the differentiation of the blood islands, and the initial formation of a well-known protein, haemoglobin (Hb), in chick embryonic tissues. After de-embryonation, chick blastoderms, from the stage of primitive streak onwards, can be cultured in vitro on a saline agar medium supplemented with glucose.


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