REGULATION OF THE INITIATION OF HEMOGLOBIN SYNTHESIS IN THE BLOOD ISLAND CELLS OF CHICK EMBRYOS: I. QUALITATIVE STUDIES OF THE EFFECTS OF ACTINOMYCIN AND δ-AMINOLEVULINIC ACID

1966 ◽  
Vol 44 (11) ◽  
pp. 1543-1560 ◽  
Author(s):  
S. D. Wainwright ◽  
Lillian K. Wainwright
Keyword(s):  
Aminolevulinic Acid ◽  
Basal Medium ◽  
Primitive Streak ◽  
Chick Embryos ◽  
Messenger Rnas ◽  
Stages Of Development ◽  
Linear Rate ◽  
Hemoglobin Synthesis ◽  
Naked Eye ◽  
Somite Stage

Intact and de-embryonated blastodiscs of chick embryos from all stages of development between the definitive primitive streak and the 10-somite stage were incubated on simple solid synthetic media. On the basal medium, blastodiscs at all initial stages of development synthesized hemoglobin readily visible to the naked eye within 24 hours, incorporated leucine into protein at an approximately linear rate for 24 hours, and incorporated uridine into RNA at a roughly linear rate for at least 6 hours after a short lag.Blastodiscs taken before the 1-somite stage failed to synthesize any detectable hemoglobin on medium containing 2 μg/ml of actinomycin, whereas those token at later stages synthesized hemoglobin visible to the naked eye. This concentration of actinomycin totally inhibited the incorporation of uridine into high molecular weight RNA within 2–3 hours, but the incorporation of leucine into protein was not inhibited for 6–8 hours. The residual incorporation of uridine was entirely into the soluble RNA fraction.At 10 μg/ml, actinomycin markedly inhibited the synthesis of hemoglobin by blastodiscs taken at stages earlier than the 6-somite embryo, but did not markedly affect hemoglobin synthesis by the more advanced blastodiscs. This concentration of actinomycin caused only slightly greater inhibition of the incorporation of uridine into acid-precipitable material than the smaller concentration for all blastodiscs, and was not markedly more inhibitory for the incorporation of leucine into protein.The presence of δ-aminolevulinic acid overcame the inhibitions of synthesis of hemoglobin by actinomycin but did not prevent the inhibitions of incorporation of uridine into RNA and of leucine into protein.Regulation of the onset of rapid hemoglobin synthesis appears to be at the translation level, probably through the supply of δ-aminolevulinic acid. The latter is probably regulated through synthesis of RNAs formed at the head-fold stage. Messenger RNAs for globin synthesis are present at the stage of the definitive primitive streak.

A kinetic study of the effects of δ-aminolevulinic acid upon the synthesis of embryonic and fetal hemoglobins in the blood islands of the developing chick blastodisc

1970 ◽  
Vol 48 (4) ◽  
pp. 400-406 ◽  
Author(s):  
S. D. Wainwright ◽  
Lillian K. Wainwright
Keyword(s):  
Aminolevulinic Acid ◽  
Fetal Hemoglobin ◽  
Primitive Streak ◽  
Stages Of Development ◽  
In Ovo ◽  
Rapid Synthesis ◽  
Somite Stage ◽  
Stage Of Development ◽  
Solid Media

Chick blastodiscs contained and synthesized a functional (embryonic) hemoglobin as early as the stage of the definitive primitive streak. The rate of hemoglobin synthesis in ovo rose dramatically at about the stage of the 3-somite embryo.Embryos explanted onto solid media at the 3-somite stage of development continued to synthesize hemoglobin in vitro for at least 12 h. The rate of synthesis rose markedly at the 6-somite stage of development, coincident with the onset of rapid synthesis of fetal hemoglobin. A supplement of exogenous aminolevulinic acid markedly stimulated the synthesis of hemoglobin before the 6-somite stage of development, but had little or no effect thereafter. These responses were obtained on both minimal and rich media. Levels of hemoglobin formed were characteristic of the final stages of development attained, regardless of initial stage of development, medium used, or time of incubation required to attain the final level of development.A new regulatory process resulting in elimination of a requirement for continuous presence of egg homogenate for maximal rates of hemoglobin was revealed. This took place between the 6- and 8-somite stages of development at the time of onset of rapid synthesis of fetal hemoglobin.

Influence of diandric and digynic triploid genotypes on early mouse embryogenesis

Development ◽  
1989 ◽  
Vol 105 (1) ◽  
pp. 137-145
Author(s):  
M.H. Kaufman ◽  
K.K. Lee ◽  
S. Speirs
Keyword(s):  
Embryonic Development ◽  
Primitive Streak ◽  
Stages Of Development ◽  
Mouse Embryogenesis ◽  
Somite Stage ◽  
Wide Range ◽  
Extraembryonic Membranes ◽  
Early Mouse ◽  
Hydatidiform Moles

Standard micromanipulatory techniques were used to produce tripronucleate diandric and digynic triploid mouse conceptuses. When these were transferred to suitable recipients, most implanted. A wide range of embryonic stages from the primitive streak to the 15- to 25-somite stage were isolated in both triploid series in otherwise identical recipients. In the diandric triploid series, all of the embryos recovered appeared to be morphologically normal, but considerably smaller than fertilized embryos analysed at similar stages of development. This contrasts with the digynic triploid conceptuses which, though also ranging from the primitive-streak stage to about the 10- to 15-somite stage at the time of their isolation, generally showed poorer embryonic development than the diandric triploids, and were invariably morphologically abnormal. Unlike the situation observed in man, where the placentas of diandric triploid conceptuses commonly display widespread trophoblastic hyperplasia and form the characteristic ‘partial’ or ‘incomplete’ type of hydatidiform moles, the extraembryonic membranes of the diandric triploid mouse conceptuses (as well as the digynic triploids) did not appear to be grossly abnormal).

REGULATION OF THE INITIATION OF HEMOGLOBIN SYNTHESIS IN THE BLOOD ISLAND CELLS OF CHICK EMBRYOS: V. QUALITATIVE STUDIES OF THE EFFECTS OF PROFLAVINE

1967 ◽  
Vol 45 (10) ◽  
pp. 1483-1493 ◽  
Author(s):  
S. D. Wainwright ◽  
Lillian K. Wainwright
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Developmental Stages ◽  
Low Molecular Weight ◽  
Chick Embryos ◽  
Hemoglobin Synthesis ◽  
Blood Island ◽  
Naked Eye ◽  
Stage Of Development ◽  
Eye Formation

Most blastodiscs explanted before the 1-somite stage of development onto medium containing 20 μg of proflavine/ml failed to form any detectable hemoglobin, whereas the majority explanted at later stages formed hemoglobin visible to the naked eye. Formation of hemoglobin by 2-somite blastodiscs was suppressed by proflavine at 50 μg/ml, but 6-somite blastodiscs formed visible quantities of hemoglobin. Incorporation of leucine into protein was inhibited by proflavine, and almost the same amount of inhibition was produced at both concentrations of inhibitor for all developmental stages examined.Net incorporation of uridine into acid-precipitable material was moderately inhibited by 20 μg of proflavine/ml, and almost totally suppressed by 50 μg of proflavine/ml. Only polynucleotides of low molecular weight accumulated in blastodiscs treated with either concentration of proflavine, but preformed stable RNA's were not degraded in the presence of inhibitor. Proflavine appeared to inhibit synthesis of, or cause degradation of, new RNA of high molecular weight at both concentrations. Accumulation of RNA of low molecular weight continued at a low proflavine concentration, but little additional RNA accumulated at a high proflavine concentration.

The origin and movement of prelung cells in the chick embryo as determined by radioautographic mapping

Development ◽  
1970 ◽  
Vol 24 (3) ◽  
pp. 497-509
Author(s):  
Glenn C. Rosenquist
Keyword(s):  
Chick Embryo ◽  
Primitive Streak ◽  
Lateral Margin ◽  
Chick Embryos ◽  
Process Stage ◽  
Somite Stage ◽  
Dorsal Mesentery ◽  
Area Pellucida

The origin of the prelung cells was determined by tracing the movements of [3H]thymidinelabelled grafts excised from medium-streak to 4-somite stage chick embryos and transplanted to the epiblast, streak, and endoderm-mesoderm of similarly staged recipient embryos. At the medium-streak stage the prelung endoderm cells are in the anterior third of the primitive streak; they shortly begin to migrate anteriorly and laterally into the endoderm layer. They are folded into the gut beginning at about the 4-somite stage, and begin to reach their definitive position in the ventrolateral gut wall at the 10- to 16-somite stage. At the ± 22-somite stage the prelung endoderm begins to burrow into the overlying splanchnic layer of mesoderm, pushing the prelung mesoderm ahead of it. At the medium-streak stage the prelung mesoderm is in the epiblast (dorsal) layer about half-way to the lateral margin of the area pellucida on either side of the streak, at a level about half-way between the anterior and posterior ends of the streak. From this position the prelung mesoderm migrates medially to the streak and is invaginated into the mesoderm layer at a position about half-way between the anterior and posterior ends of the streak. As a section of the dorsal mesentery, it migrates anteriorly and laterally from the streak into the splanchnic mesoderm lateral to the somites. From the head process stage to the early somite stages, the prelung mesoderm is located posterior to the prelung endoderm. The prelung mesoderm continues to migrate with the splanchnic mesoderm into the mesentery dorsal to the heart, where it invests the prelung endoderm after the 16- to 19-somite stage. Beginning at about the 22-somite stage, the prelung endoderm penetrates the prelung mesoderm and the bilateral bronchi are formed.

The origin and movements of the hepatogenic cells in the chick embryo as determined by radioautographic mapping

Development ◽  
1971 ◽  
Vol 25 (1) ◽  
pp. 97-113
Author(s):  
Glenn C. Rosenquist
Keyword(s):  
Chick Embryo ◽  
Developmental Stage ◽  
Primitive Streak ◽  
Limb Bud ◽  
Chick Embryos ◽  
Lateral Plate Mesoderm ◽  
Lateral Plate ◽  
Somite Stage ◽  
Definition Of ◽  
Exact Definition

The origin of the prehepatic cells was determined by tracing the movements of [3H]thymidine-labelled grafts excised from medium-streak to 4-somite stage chick embryos and transplanted to the epiblast, streak and endoderm-mesoderm layer of similarly staged recipient embryos. Although exact definition of prehepatic areas was not possible because of the small number of grafts placed at each developmental stage, the study showed in general that at the medium-streak stage, the prehepatic endoderm cells are in the anterior third of the primitive streak; they shortly begin to migrate anteriorly and laterally into the endoderm layer ventral to the precardiac areas of mesoderm. They are in the yolk-sac endoderm at the 2–4-somite stage, and by the 15–17-somite stage are clustered at the anterior intestinal portal. At the 26-somite to early limb-bud stages, the anterior and posterior liver diverticula have formed from these endoderm cells, and some of the branches of the diverticula may have reached the prehepatic mesenchyme, where the two tissues have begun to form cords and sinuses. At the medium-streak stage, the prehepatic mesoderm is located slightly more than halfway from the anterior to the posterior end of the primitive streak. From this position it migrates anteriorly and laterally into the lateral plate mesoderm, and from the head-process to the 2–4-somite stage it is situated posterior to the prehepatic endoderm and posterior and lateral to the heart-forming portion of the splanchnic layer. By the 15–17-somite stage the prehepatic mesoderm has reached a position in the splanchnic layer of mesoderm which forms the dorsolateral wall of the sinus venosus. By the 26-somite to early limb-bud stage the hepatic diverticula have joined with the hepatic mesenchyme to form the rudimentary cords and sinuses of the liver.

scholarly journals Vasculogenesis in the early quail blastodisc as studied with a monoclonal antibody recognizing endothelial cells

Development ◽  
1987 ◽  
Vol 100 (2) ◽  
pp. 339-349 ◽  
Author(s):  
L. Pardanaud ◽  
C. Altmann ◽  
P. Kitos ◽  
F. Dieterlen-Lievre ◽  
C.A. Buck
Keyword(s):  
Monoclonal Antibody ◽  
Endothelial Cells ◽  
Single Cells ◽  
Primitive Streak ◽  
Vascular Network ◽  
Vascular Tree ◽  
Somite Stage ◽  
Area Vasculosa ◽  
Haemopoietic Cells ◽  
Area Pellucida

QH1, a monoclonal antibody that recognizes quail endothelial and haemopoietic cells, was applied to quail blastodiscs in toto, in order to analyse by immunofluorescence the emergence of the vascular tree. The first endothelial cells were detected in the area opaca at the headfold stage and in the area pellucida at the 1-somite stage. Single cells then interconnected progressively, especially in the anterior intestinal portal and along the somites building up the linings of the heart and dorsal aortas. This study demonstrates that endothelial cells differentiate as single entities 4 h earlier in development than hitherto detected and that the vascular network forms secondarily. The horseshoe shape of the extraembryonic area vasculosa is also a secondary acquisition. A nonvascularized area persists until later (at least the 14-somite stage) in the region of the regressing primitive streak.

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.).

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