scholarly journals Tracer Studies on Chick Embryos in vitro

Development ◽  
1957 ◽  
Vol 5 (1) ◽  
pp. 51-59
Author(s):  
E. M. Pantelouris ◽  
L. Mulherkar

In the experiments to be reported here isotopic tracer techniques were combined with the techniques of chick embryo culture in vitro and of organizer transplantations. The distribution of methionine taken up under these conditions was first tested by autoradiography; methionine was also used to label organizer grafts and observations were made on the derivatives from such grafts; finally, the transfer of labelled molecules from the grafts to the induced structures was to some extent investigated. The first use of radioactive tracers to investigate the transfer of substances in induction was by Waddington (1950), employing P32 on Amphibian embryos. Our experiments can be viewed as an extension of these investigations and those of Abercrombie & Causey (1950) and Islam (1953) who first used radioactive tracers to label embryonic grafts in chick embryos and explants respectively, and of Feldman & Waddington's (1955) work on the uptake of labelled methionine by the chick 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.


1951 ◽  
Vol 94 (5) ◽  
pp. 401-413 ◽  
Author(s):  
M. Michael Sigel ◽  
Anthony J. Girardi ◽  
Emma G. Allen

Because of the peculiar properties of the psittacosis-lymphogranuloma group of viruses, the pattern of multiplication in the allantois of the chick embryo of one of their number, meningopneumonitis virus, was studied. This was done by determination of the changes in its infectivity for mice and chick embryos. Titration of infectivity in embryos proved to be a more sensitive procedure than titration in mice; the latter procedure however, had the advantage of greater simplicity and gave more clear-cut results. The mouse titration method was used in most of the experiments. Following inoculation of virus into the allantois, there was a slow decrease in infectivity in the allantoic fluids followed by an increase due to appearance of new virus between 24 and 48 hours. The slope of declining infectivity in the allantoic fluids in ovo was similar if not identical with the slope of decreasing infectivity in allantoic fluids in vitro caused by thermal degradation of virus. Multiplication of the virus in allantoic membranes was characterized by the following pattern: (a) Increase in infectivity in the first few hours (exact duration of increase depended on concentration of virus in inoculum) due to adsorption of virus. (b) Decrease in infectivity up to about 20 to 24 hours. (c) Increase in infectivity due to appearance of the new generation of virus. The growth curve of meningopneumonitis is analyzed and the pattern of growth is discussed in the light of the present concepts of viral multiplication.


1977 ◽  
Vol 164 (3) ◽  
pp. 533-539 ◽  
Author(s):  
A Oikarinen

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.


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.


Development ◽  
1980 ◽  
Vol 59 (1) ◽  
pp. 217-222
Author(s):  
Yal C. Sheffield ◽  
Robert E. Seegmiller

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.


1972 ◽  
Vol 51 (2) ◽  
pp. 707-709 ◽  
Author(s):  
J.B. Ramsey ◽  
M.A. Boone
Keyword(s):  

1937 ◽  
Vol 14 (2) ◽  
pp. 229-231 ◽  
Author(s):  
C. H. WADDINGTON

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.


Development ◽  
1973 ◽  
Vol 30 (2) ◽  
pp. 449-458
Author(s):  
A. Jurand

Teratogenic activity of methadone HCl (Physeptone, Burroughs Wellcome and Co.) was tested on inbred JBT/Jd and outbred Q strain mouse embryos and on chick embryos. 22–24 mg/kg injected subcutaneously on the 9th day of pregnancy caused by the 13th day exencephaly in 56 out of 479 JBT/Jd embryos but after 32 mg/kg only in 1 out of 220 of the Q strain. Some affected JBT/Jd embryos showed also rachischisis in the cervical area. The second abnormality shown by the embryos of both strains is Z-shaped kinkage of the spinal cord. In explanted chick embryos cultured in vitro as well as in embryos treated in ovo methadone causes non-closure of the neural tube with extensive necrosis of the neural plate cells in the cephalic region. The results of this study indicate that methadone, which is a neutropic drug, has an embryotoxic activity directed against the developing central nervous system.


1984 ◽  
Vol 224 (3) ◽  
pp. 769-777 ◽  
Author(s):  
A Ferioli ◽  
C Harvey ◽  
F De Matteis

The ability of drugs to cause uroporphyria in hepatocytes from 17-day-old chick embryos has been investigated and the response of the cells in culture compared with that of the intact liver of the embryos in ovo. In this chick-embryo system, drugs that cause accumulation of uroporphyrin within 19-24 h can only do so in culture; in contrast, 2-allyl-2-isopropylacetamide and 3,5-diethoxycarbonyl-1,4-dihydrocollidine, which stimulate production of protoporphyrin, are effective both in culture and in ovo. A role of exogenous iron in worsening drug-induced uroporphyria was demonstrated in cultures of hepatocytes; iron also caused preferential accumulation of uroporphyrin from added 5-aminolaevulinate in the absence of a porphyrogenic chemical. Uroporphyria was induced in cultures of hepatocytes by drugs of widely different structures, suggesting that the primary molecular target with which they interact may be relatively aspecific in its binding characteristics. These results are briefly discussed, and two alternative hypotheses for the drug-induced effect in uroporphyrinogen metabolism are considered.


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