Ultrastructure and functions of ring-shaped nucleoli, fibrillar centers and nucleolar vacuoles

Evidence on the ultrastructure of ring-shaped nucleoli in several cell types is reviewed. Detailed attention is paid to these structures in PEK culture cells after the action of actinomycin D, to infiltrating carcinomas of human breast after different degrees of malignization, and to hepatocytes of 11 day old chick embryos. On the basis of our own and literary data, the ring-shaped nucleoli are classified into two groups: 1) those with centrally located fibrillar centres, and 2) those containing large central vacuole. Connections of fibrillar centres with the nucleolus organizer regions in the interphase nucleolus and mitotic chromosomes are considered in detail. Literary and our own data on the genesis and functional activity of ring-shaped nucleoli with the central vacuole, as well as nucleoli with different degrees of vacuolization (vacuolized nucleoli) are specially analyzed. Cellular function, properties and significance of the central vacuole in low differentiated cells are discussed.     

RNA synthesis in the circular nucleoli of hepatocytes]

By analysis of serial sections it has been revealed that the so-called ring-like nucleoli of hepatocytes consist of a cavity with an amorphous contents surrounded by fibrillar and granular material. Such nucleoli are sometimes encountered in normal animals; the number of ring-like nucleoli increases considerably in chronic pathological process caused by repeated CCl4 injections. The capacity of RNA synthesis in the ring-like nucleoli was revealed by means of electron-microscopic autoradiography.     

Ultrastructure of isoproterenol-induced myocardial damage in chick embryos

Isoproterenol is known to cause severe myocardial lesions when given in toxic doses to adult homoiotherms. Previous studies on chick embryos revealed myocardial damage with scattered necroses in the outer layer of ventricular myocardium. The present ultrastructural study, performed on embryos 6 to 20 days old, has shown various types of cellular lesions; mainly cellular oedema, mitochondrial swelling, necroses of isolated cardiac muscle cells, fatty degeneration, accumulation of glycogen, and signs of increased proteosynthesis in the surviving muscle cells. Morphological features of the lesions differed from those which are known to be induced by isoproterenol in adult animals and seemed to depend on the stage of embryonic development.     

Analysis of ring-shaped nucleoli in serially sectioned human lymphocytes

Summary Serial section analysis has demonstrated that ring-shaped nucleoli of mature human lymphocytes are spherical structures consisting of a peripheral ribonucleoprotein shell that surrounds one large fibrillar center. The shell exhibits usually one or, less frequently, two openings. The fibrillar center is in contact with the nucleoplasm and perinucleolar condensed chromatin, which frequently appears as a pedicle-like structure. Several chromocenters are associated with the ring-shaped nucleolus.     

Morphological studies on ring-shaped nucleoli and micronucleoli in frog erythroblasts

Frog erythroblasts and erythrocytes were studied to provide more information on the incidence of ring-shaped nucleoli and micronucleoli during cell maturation and differentiation.     

Hepatotropism of Tyzzer bacteria in experimentally infected chick embryos]

Tyzzer's organism from mice propagated more remarkably in hepatocytes than in yolk-sac epithelial cells producing confluent necrosis of the liver after intravenous as well as intravitelline inoculation. Some lesions with bacterial growth were also produced in the heart muscles and kidneys. The organisms from mouse, rat, hamster and kitten were shown to be equally pathogenic for chick embryos.     

Fate of amplified nucleoli in Xenopus laevis embryos

We have followed the fate of two components of extrachromosomal nucleoli, amplified ribosomal DNA (rDNA) and 7.5 kb precursor rRNA, during early embryogenesis of Xenopus laevis. Other workers have shown that the amount of amplified rDNA accumulated during oogenesis remains unchanged through the 16-cell stage of embryogenesis. Here we show that as embryonic cleavage continues, the amount of amplified rDNA decreases until it is no longer detectable in the early gastrula embryo. In contrast, the amount of 7.5 kb precursor rRNA in eggs, early cleavage stage embryos, or blastula stage embryos is the same as in oocyte nuclei. Since no rRNA synthesis occurs during these early stages, we conclude that the precursor rRNA sequences synthesized in the oocyte are neither processed nor degraded during early development. The amplified rDNA is not replicated in the early embryo even though the chromosomal DNA of the embryo replicates every 30 min during the first 7.5 hr of embryogenesis. When amplified rDNA is purified and then injected into cleaving embryos, however, we find that it is replicated. This finding suggests that some factor(s) prevents the endogenous amplified rDNA from responding to the cellular replication signals. We show that methylation of cytosine in the rDNA is not related to the DNA's capacity for replication in this system since amplified (unmethylated) and chromosomal (methylated) rDNA are both replicated when injected into embryos. The methylation pattern of these rDNAs appears to be maintained after replication in the embryo.     

Germ cell cycles during sex inversion in chick embryos]

A study was made of the germ cell cycles of 11 days old embryos injected male or female hormones on the 4 th day of incubation. The cell cycles duration in genetically male 11 days embryos treated with esradiol-benzoate was close to that registered in oogonia of both treated and non-treated female embryos of the same age. The testosterone propionate injection caused an acceleration of the genetically male sex cell proliferation and a decrease of the reproduction rate of the female sex cells. It is proposed that under normal conditions female sex hormones inhibit a hypothethic factor that determines the decrease of cell proliferation during the male embryo development.     

Ultrastructure and behavior of the nucleoli and fibrillar centers during cell differentiation of liver erythroblasts of 12-day-old mouse embryos

The transformation of nucleolus and its structural components in the main groups of erythroid cells (from pronormoblasts to reticulocytes and dividing ones) has been studied. It is shown that during inactivation of the nucleolus, the granular component is reduced, and the degree of chromatin condensation increases. Enlargement and "naking" of fibrillar centres are also observed. At the stage of basophilic and polychromatophilic erythroblasts, the nucleolus has a mushroom-like shape with well developed fibrillar centres, which lie at the border of the nucleolus. Nucleolar RNP components consist predominantly of a fibrillar component and forms "caps" of these mushroom-like structures. Therefore, at this stage "free" fibrillar centres are found on ultrathin sections, if the section plane runs only through the fibrillar centre, or through ring-shaped nucleoli, i.e. the fibrillar centre surrounded by sheet of nucleolar RNP fibrilles, when the mushroom-like nucleolus is cut tangentially. Using serial section technique, small round nucleoli with an extremely weakly developed RNP material or free fibrillar centres, resembling those in telophase nuclei, are shown on the terminal stage of nucleolus transformation. It is noted that the main groups of erythroid cells differ from each other not only in the chromatin condensation degree, but also in the development of nucleolus material and in the size of fibrillar centres. However, such differences exist in either cell group. Consequently, we can distinguish between cell populations being on different stages of maturation. On this basis, we described on intermediate population of cells, which possess signs of pronormoblasts and basophilic erythroblasts. In all the cases, strands of electron opaque material bound with the condensed chromatin are present in fibrillar centres.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphometric analysis of neural tube cells in neurulating chick embryos]

Preliminary report on a morphometric study of the neuroepithelium of 1 S and 7 S -- stage chick embryos. We show first that the nucleo-cytoplasmic ratio and then, that the volume fraction of intercellular spaces, nuclei and cytoplasm as they relate to the neural tube, differ significatively from cephalic to caudal portion of the neural tube. At stage 7 S only the volume fraction of intercellular spaces, nuclei and cytoplasm in the neural tube change along the cephalo-caudal axis of the embryos. Finally, between the two stages, at the back of the head, 1) volume fraction of intercellular spaces in the neural tube remains unchanged 2) nucleo-cytoplasmic ratio increase 3) volumetric density and size of mitochondria do not change, while surface density of endoplasmic reticulum in cytoplasm increases.     

Respiration and glycolysis in the liver of developing chick embryos and chicks]

Oxygen uptake in liver slices remains constant between the 12th and the 17th days of embryonic development, being equal to that in 30-60-day chicks. During the transition from allantoic respiration to the pulmonary one, oxygen consumption decreases, the decrease being observed up to the end of embryonic period. After hatching, oxygen consumption increases 4-5-fold to the 6-7th and decreases up to the initial level at the 10th day. Respiration of mitochondria isolated from the liver and concentration of cytochromes in mitochondria remain constant. The value P/O is the lowest, whereas catalase activity is the highest during hatching. The intensity of anaerobic glycolysis changes similarly to that of respiration.     

Regulation of sugar content in the blood during artificial hyperglycaemia in chick embryos]

Intravenous injections of glucose (8.6 mg/ml of blood) have been made 9-18 day chick embryos, 1-day chicks and adult hens. Glucose content of the blood was measured 15 and 30 min., 1,4 and 8 hours after the injection. It was shown that already 9-day embryos possess a regulatory system which restores the initial glycaemic level. During the development of the organism, the efficiency of this system increases. The data obtained may be used in theoretical studies on the sugar system of the blood.     

The isolation of nucleoli from ungerminated pea embryos

A method is described for the isolation of nuclei and nucleoli from ungerminated pea embryos. Electron micrographs of the nucleolar preparation indicate a high degree of purity but only a partial preservation of composition, because of a loss of material from the nucleoli which occurs during the isolation procedure.     

Calcium regulation in the embryonic chick. II. Ultrastructure of the parathyroid glands in shell-less and in ovo embryos

The ultrastructure of the parathyroid glands was studied in chick embryos developing normally in ovo or in shell-less culture (after removal of the eggshell). Shell-less chick embryos are significantly hypocalcemic relative to their in ovo counterparts. At 12 days of incubation, the parathyroid glands of shell-less embryos contain more lipid and show evidence of increased protein synthetic activity relative to those grown in ovo (more rough endoplasmic reticulum, presence of some dense secretory granules). The glands from in ovo embryos do not contain secretory granules at this age. At 15 days of incubation, the in ovo glands have developed signs of protein synthetic activity similar to those of the 12-day shell-less embryos. However, the parathyroids of the 15-day shell-less embryos appear strikingly more active than at 12 days, containing stacks of concentric RER membranes and increased numbers of secretory granules. By 18 days of incubation, the ultrastructure of the glands of the two groups is indistinguishable, both appearing to be more active than the 15-day shell-less group. Thus, protein synthetic activity of the parathyroid glands, as detected by ultrastructural alterations of the chief cells, normally appears to be initiated during the latter part of embryogenesis (by approximately 15 days incubation) and its onset can be stimulated at least 3 days prematurely by hypocalcemia.     

Immunofluorescence study of the formation of evolutionary stable lens proteins in chick embryos]

In the lens of fishes (carp, spiny dogfish) beta-crystallins were identified which were characteristic also of reptiles, amphibians, birds and mammals (evolutionary stable beta-crystallins). The dynamics of the formation of such beta-crystallins in 5--14 days old chick embryos was studied by the indirect immunofluorescence method with antisera to fish lens. These proteins are reliably indentified first at the lens sections from 7--8days old chick embryos. At all stages under study these beta-crystallins are localized mainly in the epithelial cells and practically not found in the lens fibers. They were, however, found in the fibrous (central) part of developing lens as well by the method of immunoelectrophoresis.