Synthesis of globin chains in the erythropoietic sites of the early chick embryo

Development ◽  
1983 ◽  
Vol 77 (1) ◽  
pp. 153-165
Author(s):  
L. Fucci ◽  
C. Cirotto ◽  
L. Tomei ◽  
G. Geraci

The synthesis of globins in the chick embryo before the onset of circulation has been studied in situ by specific immunofluorescence labelling of embryonic sections and by labelling newly synthesized proteins in ovo and in vitro in embryonic explants with [3H]leucine. The presence of major primitive haemoglobins is observed by 28 h of incubation. The minor primitive haemoglobins become detectable by immunofluorescence after 40 h of development, shortly before the onset of circulation. 3H-labelling shows that one definitive α chain is synthesized, though in low concentration, from the initial globin detection. The other definitive α chain is observed in embryos of at least 40 h of development. The relative concentration of the two definitive α chains changes rapidly with development indicating a specific mechanism of regulation. An erythropoietic site is observed in the wall of the dorsal aorta in embryos of about 45–50 h of development. From the initial detection, those cells contain all four primitive embryonic haemoglobins, in contrast to what is observed for the cells of the blood islands.

Development ◽  
1959 ◽  
Vol 7 (1) ◽  
pp. 66-72
Author(s):  
L. Gwen Britt ◽  
Heinz Herrmann

The recent development of techniques originally devised by Waddington (1932) for the maintenance of the explanted chick embryo (Spratt, 1947; New, 1955; Wolff & Simon, 1955) has opened the possibility of determining quantitatively some parameters of the developmental processes occurring in embryonic tissues under these conditions. As a result of such measurements, protein accumulation in explanted embryos was found to be much smaller than in embryos developing in the egg. On the other hand, the progress of somite formation was found to take place at similar rates in embryos developing as explants or in situ (Herrmann & Schultz, 1958). The slow rate of protein accumulation in the explanted embryos made it seem desirable to investigate whether under some other conditions of explantation protein accumulation would approach more closely the rate of protein formation observed in the naturally developing embryo.


Development ◽  
1960 ◽  
Vol 8 (4) ◽  
pp. 369-375
Author(s):  
P. H. S. Silver

It seems to be generally accepted that experimenting in ovo on the chick during the early stages of development (up to about 48 hours) is fraught with the greatest difficulty. After about this time no serious technical problems arise and a high proportion of successful results can be expected. It is natural to ask why there should be this change-over from extreme difficulty to reasonable simplicity. New (1955) attributed to this ‘inaccessibility of the chick embryo in the egg’ the invention of his own and many other in vitro methods during the last 30 years. There is no doubt that, when short-term experiments only are required, in vitro methods will probably always be preferred. But all in vitro methods suffer from the disadvantage that the embryo cannot be expected to survive for more than 48 hours or so after explantation.


2012 ◽  
Vol 302 (1) ◽  
pp. F29-F37 ◽  
Author(s):  
Silvia Medrano ◽  
Maria C. Monteagudo ◽  
Maria Luisa S. Sequeira-Lopez ◽  
Ellen S. Pentz ◽  
R. Ariel Gomez

We have shown that microRNAs (miRNAs) are necessary for renin cell specification and kidney vascular development. Here, we used a screening strategy involving microarray and in silico analyses, along with in situ hybridization and in vitro functional assays to identify miRNAs important for renin cell identity. Microarray studies using vascular smooth muscle cells (SMCs) of the renin lineage and kidney cortex under normal conditions and after reacquisition of the renin phenotype revealed that of 599 miRNAs, 192 were expressed in SMCs and 234 in kidney cortex. In silico analysis showed that the highly conserved miR-330 and miR-125b-5p have potential binding sites in smoothelin ( Smtn), calbindin 1, smooth muscle myosin heavy chain, α-smooth muscle actin, and renin genes important for the myoepithelioid phenotype of the renin cell. RT-PCR studies confirmed miR-330 and miR-125b-5p expression in kidney and SMCs. In situ hybridization revealed that under normal conditions, miR-125b-5p was expressed in arteriolar SMCs and in juxtaglomerular (JG) cells. Under conditions that induce reacquisition of the renin phenotype, miR-125b-5p was downregulated in arteriolar SMCs but remained expressed in JG cells. miR-330, normally absent, was expressed exclusively in JG cells of treated mice. In vitro functional studies showed that overexpression of miR-330 inhibited Smtn expression in SMCs. On the other hand, miR-125b-5p increased Smtn expression, whereas its inhibition reduced Smtn expression. Our results demonstrate that miR-330 and miR-125b-5p are markers of JG cells and have opposite effects on renin lineage cells: one inhibiting and the other favoring their smooth muscle phenotype.


1994 ◽  
Vol 190 (1) ◽  
pp. 141-153
Author(s):  
M J Packard

Two explants of shell were removed from each of several fertile eggs of domestic fowl at different times during incubation. The chorioallantoic membrane (CAM) was removed from one of the explants (SHELL ONLY) and was left in situ on the other (SHELL+CAM). Explants were cultured for 24, 48 or 96 h at 37 degrees C and 5% CO2 in air in individual Petri dishes containing Dulbecco's modified Eagle's medium, bovine serum albumin, penicillin and streptomycin. Both SHELL+CAM and SHELL ONLY explants released calcium into the culture medium, but the former released considerably more calcium than the latter. More calcium was released by SHELL+CAM explants taken from older eggs than from younger ones, but the age of the donor eggs did not affect release of calcium by SHELL ONLY explants. In addition, release of calcium by SHELL+CAM explants exceeded that shown by SHELL ONLY explants for multiple 24 h intervals. However, the capacity for sustained release of calcium by SHELL+CAM explants declined with age and maturity of the CAM. Manipulations that lead to the death of the CAM abolish the capacity for SHELL+CAM explants to release more calcium than SHELL ONLY explants. Differential release of calcium by SHELL+CAM explants was not attributable to calcium present in the CAM at the onset of culture or to non-specific degradation of the shell by intracellular constituents released as a result of the death of the CAM. Taken in concert, these results indicate that the CAM mobilizes calcium from the eggshell during in vitro culture.


1989 ◽  
Vol 92 (3) ◽  
pp. 497-504 ◽  
Author(s):  
E.J. Sanders ◽  
S. Prasad

At the time of gastrulation in the chick embryo the upper epiblast layer penetrates its own basement membrane at the primitive streak so that its cells may invade the underlying tissue space. In so forming the primary mesoderm, the cells undergo a concomitant epithelial-to-mesenchymal transformation. In this study, epiblast tissue has been explanted onto a basement membrane gel in order to examine its invasive potential. Fully ingressed primary mesoderm cells were able to penetrate the gel as individual cells, during the course of which the texture of the gel was disrupted. By contrast, epiblast tissue taken from the immediate vicinity of the primitive streak penetrated the gel, but only as a coherent tongue of cells and without gel disruption. These tongues of cells did not undergo the epithelial-to-mesenchymal transformation, and consequently spread as a epithelial sheet when replated on glass. Thus, the absence of gel disruption correlated with the failure of transformation, suggesting that these two events may be linked and that they may require in situ cell interactions for their manifestation. Tissue from the lateral epiblast failed to penetrate the gel. Instead, this tissue either spread on the gel surface or rounded up into a hollow sphere with the basal surface of the cells innermost. In the former case, despite the cell spreading, no lamina densa was organized beneath the sheet, but in the latter case polarity reversal occurred with the formation of a new lamina densa at the cell-gel interface.(ABSTRACT TRUNCATED AT 250 WORDS)


2019 ◽  
Vol 7 (18) ◽  
pp. 3106-3109 ◽  
Author(s):  
Massimo Fioranelli ◽  
Alireza Sepehri ◽  
Maria Grazia Roccia ◽  
Chiara Rossi ◽  
Petar Vojvodic ◽  
...  

BACKGROUND: Recently, some new methods for sexing of chicken eggs by fluorescence and Raman spectroscopy through the shell membrane have been proposed. On the other hand, in another investigation, a new virus medical imaging has been suggested. AIM: In this research, summing over these considerations, a new technique for sexing of chicken eggs by virus spectroscopy through the shell membrane is proposed. METHODS: (Please, add text …) RESULTS: (Please, add text …) CONCLUSION: It is shown that viruses outside the shell of egg can communicate with materials inside it and determine the gender of chick embryo and its evolutions.


1986 ◽  
Vol 82 (1) ◽  
pp. 73-84
Author(s):  
R.S. Tuan ◽  
M.J. Carson ◽  
J.A. Jozefiak ◽  
K.A. Knowles ◽  
B.A. Shotwell

During chick embryonic development, the chorioallantoic membrane (CAM) is responsible for the mobilization of shell calcium into the embryonic circulation. The calcium-transport function of the CAM was studied here by measuring CAM calcium uptake in vivo and in vitro. The in vivo technique involved the use of an uptake chamber constructed on top of the CAM in situ. The in vitro methods included two systems: CAM tissue disks and cell-free microsomal membranes isolated from the CAM. Analyses using these three assays show that calcium uptake by the CAM exhibited characteristics indicative of active transport, such as temperature dependence, saturability, energetic requirement and ion specificity. The data also show that calcium-uptake activities of the CAM increase as a function of embryonic age in a manner coincident with the increased accumulation of calcium by the developing embryo in ovo.


Development ◽  
1981 ◽  
Vol 61 (1) ◽  
pp. 175-190
Author(s):  
K. Palén ◽  
L. Thörneby

Chick embryos were treated in ovo and in vitro with L-phenylalanine from the intermediate streak stage (Hamburger & Hamilton stage 3, 12–13 h of incubation) to the 7-somite stage (H & H stage 9, 29–33 h of incubation). Treatment in ovo resulted in a large number of embryos developing somite blocks, i.e. imperfectly segmented somites. In embryos treated at an early developmental stage (12–21 h of incubation), the blocks of unsegmented somite mesoderm occurred mostly in the somite pairs 1–5, whereas treatment that began at a later stage (24–30 h of incubation) caused blocks in the somite pairs 5–10, i.e. the appearance of blocks of unsegmented somite mesoderm is correlated in time with the onset of the treatment. No difference regarding mitotic indices could be distinguished between normally segmented somites and blocks of unsegmented somite mesoderm. Autoradiography based on tritiated L-phenylalanine showed no regional differences in labelling of the chick embryo body. Electronmicroscopical observations indicate a slightly suppressed formation of microvilli in the cells of the unsegmented mesoderm blocks compared with cells in normally segmented somites. The observed disturbances are probably caused by a suppressed yolk granule decomposition in the developing somite cells. The experiments in vitro support the findings in the in ovo material; at the same time, they reveal an unexpectedly slow diffusion of L-phenylalanine through the vitelline membrane.


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