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Secretion of the anti-müllerian hormone by the gonads of experimentally sex reversed female chick embryos.

The gonads of genetically female chick embryos experimentally transformed into testes under the influence of a embryonic testis graft are able to induce in vivo the regression of Müllerian ducts when they are grafted to female embryonic hosts. On the other hand female gonads only transformed into ovotestes are ineffective on the host MDs, as in the case for female gonads. These results show that totally sex reversed gonads have the same properties as a normal testis. In particular they produce the anti-Müllerian hormone, whereas partially reversed gonads do not or only do at a very low level.

Animals↗

Protein composition of cerebrospinal fluid in the developing chick embryo.

Proteins from cerebrospinal fluid of chick embryos at 4-19 days of development were qualitative and quantitatively analyzed. The total protein concentration and the relative concentration of each protein fraction were calculated for each day. The main proteins found along development were immunoglobulin G, transferrin, alpha-fetoprotein, serum albumin, ovalbumin, prealbumin, and an unidentified component. alpha-Fetoprotein was found to be the major protein at 4-16 days, and serum albumin at 17-19 days of development. The unidentified fraction was present from 4 to 11 days; at day 5 it represented 28% of the total cerebrospinal fluid protein concentration.

Albumins↗

Immunoferritin studies on the multiplication of Sindbis virus in chick embryo fibroblasts.

Replication of Sindbis virus chick embryo fibroblasts was studied by means of the immunoferritin technique. Virus specific antigens, diffusely distributed in the cytoplasm, were observed at first 3 hours p.i. At the same time in ohter cells ferritin particles in linear arrangement occured at the surface of vesicles. At these places heavily labelled nucleocapsids were observed later on. This means, that the surface of vesicles is a site of assembly of nucleocapsids. Maturation of virus takes place at the cell surface as well as at the vesicles by budding. In early stages of replication maturation at the membrane prevalis.

Animals↗

Further studies on the size and composition of the chick embryo fibroblast cytosolic DNA complex.

The chick embryo fibroblast cytosolic DNA complex shows anomalous elution behaviour on agarose gel column chromatography. The indicated molecular size varies between 5 X 10(5) dalton (higher exclusion limit gels) and 1.4 X 10(6) dalton (lower exclusion limit gels). Chromatography on lower exclusion limit gels shows the [3H]thymidine labelled (DNA) complex as a sharp peak, coincident with a peak of [3H]uridine and [3H]lysine labelling and similar pulse labelling patterns for the three precursors but with DNA labelling lagging behind RNA and protein. Both cultured and uncultured cell cytosols show an A260 peak coincident with the [3H]precursor labelling peaks.

Animals↗

Hydromineral balance in the chick embryo: effects of hypophysectomy.

The developing chick embryo exhibits a regular pattern of accumulation and reabsorption of fluid in its extraembryonic compartments. Fluid volume (measured by dye dilution) peaks at Day 14 in both the allantois (12.4 ml) and the amnion (5.1 ml). The standard experimental procedure of removing 2 ml of albumen at 33-36 hr of incubation does not alter this pattern, but leads to a maximum decrease in fluid volume of ca. 1 ml in both amniotic and allantoic compartments. Although [Na] and [Cl] remain essentially constant in the serum, they steadily decrease in the allantois (remaining substantially below serum levels). In the amnion, [Cl] is maintained above that of serum, whereas [Na] falls below serum [Na] by Day 14. In the Day 12 (stage 38) embryo, hypophysectomy (HX) causes a substantial increase in the allantoic fluid volume and [Na] and [Cl], thus resulting in a 166% increase in total Na and Cl over that of sham-operated embryos. By contrast, HX did not alter amniotic fluid volume, although it did decrease [Na] and to a lesser degree, [Cl]. Serum [Na], but no [Cl], was reduced by HX. HX abolished the normally steep ionic gradients maintained between the allantois and both serum and amnion. Thus, the most dramatic effects of HX at Day 12 are on the allantois, which accumulates both Na and Cl at the expense of the amnion and serum. This suggests that pituitary hormones may normally act by Day 12 to reduce urinary volume and electrolyte excretion and/or to increase fluid and electrolyte reabsorption from the allantois.

Allantoin↗

[Preservation of the bipolar shape and contact orientation by mitotically dividing chick embryo myoblasts in a monolayer culture].

Chick embryo skeletal myoblasts in monolayer cell culture may by spherical during mitosis, as is commonly the case with cells in vitro, or maintain bipolar spindle-form shape throughout all the phases of mitosis. The percentage of spindle-form metaphase myoblast increases from 32% (of al the metaphases), on the 2nd day of culturing, to 80%, on the 5th day. The long axis orientation of mitotic spindle in dividing elongated myoblasts is the same as that of the whole cell. The myoblasts contact orientation (along parallel grooves of artificial substrate or along myotubes) is maintained in bipolar mitotic cells throughout mitosis and persists in daughter cells.

Animals↗

Cell lineage restrictions in the chick embryo hindbrain.

During development of the chick embryo, early neuronal differentiation and axonogenesis in the hindbrain follow a segmented pattern in register with the segmented morphology of this region. Cell marking experiments have shown that the segments, or rhombomeres, are lineage-restriction units each constructing a defined piece of the hindbrain. This raises the interesting possibility that, as in the developing fly, metamerism is used to generate level-specific anatomical structures with great and reliable precision. In the hindbrain, as for many invertebrates, lineage ancestry may be important in the determination of cell fate. The segmentation seen in this body region could therefore reflect a similar condition once present in the ancestor common to vertebrates and invertebrates.

Animals↗

Carbohydrate moieties of procollagen: incorporation of isotopically labeled mannose and glucosamine into propeptides of procollagen secreted by matrix-free chick embryo tendon cells.

Cells obtained from chick embryo tendons incorporate isotopically labeled glucosamine and mannose into the pro-alpha1 and pro-alpha2 chains of procollagen as judged by sodium dodecyl sulfate-gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The label was further localized to the propeptides of pro-alpha1 and pro-alpha2 by its chromatographic behavior after digestion with bacterial collagenase or alpha-chymotrypsin. Carbohydrate analysis of isolated pro-alpha chains showed the presence of labeled galactosamine in addition to mannose and glucosamine. Resistance to mild alkaline hydrolysis suggested that greater than 90% of the oligosaccharide units are not linked to the propeptide backbone by either serine or threonine.

Animals↗

Stimulatory role of glial cells on the differentiation of dissociated chick embryo encephalon cells.

Dissociated cells from chick embryo encephalon of 36 h of incubation have been cultivated on fibroblast, meningeal cell, or glial layers. Within the first 24 h of cultivation, cells adhered to the preformed layers more firmly than to plastic or collagen surfaces. After 48 h of cultivation, the brain cells cultured on meningeal cell or fibroblast layers behaved very similarly to those cells cultured on plastic surfaces. But when cultured on glial layers for 3 days, the brain cells differentiated into characteristic neuroblasts, as demonstrated by specific neuronal stainings. These results support the hypothesis of a stimulatory role of glial cells in neuronal differentiation. The possible causes of this stimulatory action are discussed.

Animals↗

Development of heart rate irregularities in chick embryos.

Heart rate (HR) irregularities in chick embryos were defined as large fluctuations (>10 beats/min) comprising irregular, brief deceleration and/or acceleration of instantaneous HR (IHR). IHR was determined directly from the arterial blood pressure while adequate gas exchange was maintained through an eggshell and chorioallantoic membrane. Five embryos were examined on each day from day 11 to day 19 of incubation. Baseline HR was stable until day 12-13, and on around day 13-14 transient, rapid deceleration of HR (termed V pattern) began to appear, with a subsequent increase in its frequency and magnitude. The acceleration patterns (lambda, avian omega, and periodic patterns) appeared later, and the IHR became increasingly irregular, with additional, spontaneous deceleration and acceleration patterns toward hatching. Additional experiments with intravenous administration of autonomic drugs clearly showed that rapid deceleration of HR was mediated by parasympathetic nervous function but did not always show clear relations of sympathomimetic and sympathetic blocking agents to the acceleration patterns.

Allantois↗

Release of acetylcholine by chick embryo heart before innervation.

1. In chick embryo hearts, 3-day-old and not yet innervated, repetitive direct stimulation causes a transitory inhibition of the spontaneous rhythm.2. The degree of post-stimulation inhibition depends on the frequency and duration of the artificial stimulation and on the concentration of K and Ca ions in the extracellular solution.3. After treatment with atropine (10(-5) g/ml.) post-stimulation inhibition is no longer observed. The spontaneous rhythm is accelerated by atropine. The findings therefore suggest that an ACh-like substance is released from the non-innervated embryonic heart during activity.4. By use of the dorsal muscle of the leech for biological assay the liberation of an ACh-like substance from the non-innervated embryonic heart was confirmed.

Acetylcholine↗

Evidence for contact stimulation of growth of homologous cells in superinoculated cultures of chick embryo cells.

The secondary cultures of chick embryo cells were suspended and transferred to homologous cell cultures. Cell adhesion and proliferation were studied in these superinoculated cultures. It was shown that added cells soon adhered to the underlying cell layer which results in a prompt increase in culture density followed by the activation of DNA synthesis and cell division. Stimulation of cell proliferation involved both cell subpopulations composing the superinoculated culture: cells seeded on the built-up cell layer and cells of the layer. The contact nature of added cell mitogenic action on overlaid cell proliferation was evidenced. The cell system described can be used to investigate the adhesive properties of the cell layer apical surface, the relationship between cell growth rate and culture density, and the contact stimulation of cell proliferation.

Animals↗

Amino acid and glucose transport in sarcolemmal vesicles from chick embryo heart.

Purified plasma membranes from chick embryo heart are shown to retain several functional properties of intact cardiac cells. (1) Muscarinic cholinergic receptors copurify with previously used cell-surface markers, i.e., (K+-dependent) p-nitrophenylphosphatase and insulin receptors (Paris, S., Fosset, M., Samuel, D. and Ailhaud, G. (1977) J. Mol. Cell. Cardiol. 9, 161-174). (2) Neutral amino acids (L-alanine and alpha-aminoisobutyric acid) are actively transported into these osmotically active vesicles when an Na+ electrochemical gradient is imposed. The affinity and specificity for amino acids are similar to those described for intact cardiac cells. (3) D-Glucose is taken up more rapidly than L-glucose. The sterospecific transport system is saturable and Na+-independent. The Km value is close to that observed in intact cells with glucose analogues.

Alanine↗

The development of the irido-corneal angle in the chick embryo.

The iridocorneal angle of the chick embryo was examined by scanning electron microscopy from the 7th day to the 19th day of development. The mesodermal tissue situated between the corneal endothelium and the iris differentiated into trabecular cells. These cells formed a strip on the 9th day and enlarged on the 13th day to form long cellular fibres. The trabecular meshwork was recognizable on the 15th day and reached full development only on the 19th day. The development of the trabecular meshwork was probably much earlier than is usually described. It began with differentiation of the corneal endothelial cells and these cells acquired a multipolar cell body with long cellular processes. The last stage is probably regulated to avoid an abnormal development of the angle structure. These results can be correlated with those obtained in the morphological examination of the structure of the angle in congenital glaucoma and the "cleavage syndrome" of the anterior chamber.

Animals↗

Insulin and IGFs induce apoptosis in chick embryo retinas deprived of L-glutamine.

In chick embryo retinas, cultured in serum-free medium lacking L-glutamine, IGF-I, IGF-II and insulin induced apoptotic DNA fragmentation and cell death, IGF-I being the most efficacious compound. The apoptotic effect, which was particularly evident in retinas removed from 7-day-old chick embryos, declined with the age of the embryos and disappeared after day 11. Apoptosis appeared after a time lag of 8 h and then increased with time up to 16 h. Cycloheximide, an inhibitor of protein synthesis, was capable of entirely abolishing apoptotic cell death. The effect induced by IGFs or insulin was suppressed by the addition of glutamine. Cytokine-mediated apoptosis was also observed after withdrawal of phosphate. We suggest that IGFs or insulin may produce, in retinas cultured in medium lacking L-glutamine or phosphate, a conflict of signals that could be lethal for retinal cells.

Journal Article↗

Epithelial-mesenchymal interactions in the outgrowth of limb buds and facial primordia in chick embryos.

The facial primordia in the chick embryo begin as rounded swellings that surround the primitive mouth and these grow out to form the beak. The control of proximodistal outgrowth is not well understood but may involve similar mechanisms to the limb bud. In order to test this hypothesis, combinations were made between epithelium and mesenchyme from facial primordia and limb buds. Signals from all three types of facial mesenchyme (frontonasal mass, mandibular, and maxillary) maintained the thickened apical ectodermal ridge of limb epithelium for up to 48 h. Combinations of tissues from the frontonasal mass mesenchyme and limb epithelium underwent substantial and correct morphogenesis. In contrast, poor development was observed in combinations with mandibular mesenchyme. Signals from frontonasal mass epithelium promoted outgrowth and morphogenesis of limb mesenchyme whereas mandibular and maxillary epithelium did not support joint morphogenesis. The results suggest that signals employed in the epithelial-mesenchymal interactions in facial primordia are similar but not identical to those signals used in the limb bud.

Animals↗

The effects of dexamethasone on the differentiation and the fertilisation of the germinal primordium in the chick embryo.

We showed that, in the chick embryo, the fertilisation of the attractive germinal epithelium by primary germ cells can be represented by a three-dimensional diagram in which the space and time co-ordinates are graduated in terms of the segmentation of the axial and paraxial mesoderm. We thus established that the differentiation of the coelomic epithelium into an attractive germinal epithelium and the fertilisation of the gonadal primordium both occur by mechanisms that are tightly linked to somitogenesis. In the continuous presence of a constant concentration of Dexamethasone, a marked inhibition of the rate of fertilisation of the gonadal primordium was observed. A mathematical analysis of the mode of action of the inhibitor revealed the progressive establishment of a competition between Dexamethasone and the molecule(s) responsible for the process of attraction. Given the chemical nature of the inhibitor, we propose that the endogenous factor that triggers the first step of the differentiation of the germinal primordium is a steroid-containing complex.

Animals↗

The effect of LSD on the histology and ultrastructure of the neuroepithelium of young chick embryos: a stereological study.

Five control chick embryos and fifteen exposed to LSD at concentrations of 2.5, 12.5 and 50 mug/ml were examined. The morphometric analysis performed at the light microscope level shows that (1) the section area of the neural tube increases with the two highest doses of lysergic acid diethylamide (LDS) employed, (2) the volume fraction of the intercellular spaces decreases with the two highest doses of LSD, (3) the volume fraction of the nuclei in the neural tube is not modified with any concentrations of LSD employed, (4) the volume fraction of the cytoplasm in the neural tube increases with all three concentrations, and (5) the nucleo-cytoplasmic ratio decreases with all three doses employed. Moreover, at the ultrastructural level, it was found that (1) the volume fraction of mitochondria in the cytoplasm decreases at doses of 12.5 and 50 mug/ml (2) the surface to volume ratio of the mitochondria is unchanged with any of the concentrations of LSD employed and (3) the surface density of the endoplasmic reticulum in the cytoplasm increases only with a dose of 2.5 mug/ml of LSD.

Animals↗