A radioautographic study of protein synthesis in loach blastoderm]

The dynamics of protein synthesis in the loach embryos has been studied by means of autoradiography at the stages of cleavage, blastula and gastrula. During the synchronous cleavage divisions, nuclear proteins are mainly synthesized. From the early blastula stage until the early gastrula stage, the intensity of nuclear protein synthesis increases 2.5 times whereas the intensity of cytoplasmic and total protein synthesis is low and relatively constant. After the onset of gastrulation the intensity of nuclear and cytoplasmic protein synthesis increases 3-4 times and at the late gastrula stage it decreases twice as compared with that at the midgastrula stage. During blastulation, no regional differences in the intensity of nuclear and cytoplasmic protein synthesis were found. With the onset of gastrulation, a vegeto-animal gradient of labeled aminoacid incorporation into nuclear and cytoplasmic proteins appears. During gastrulation, reliable differences were found between the intensity of labeled aminoacid incorporation into proteins of the cells of intact and dissociated blastoderms. During this period, the intensity of protein synthesis in embryonic shield is higher than that in the extraembryonic part of blastoderm.     

An electron microscopic radioautographic study of the viability of human cells after death. The postmortem activation of RNA synthesis in neutrophils]

RNA synthesis in human neutrophils isolated from blood 2.5-67 hours after death of patients was studied by means of light and electron microscopy radioautography. The rate of RNA synthesis in neutrophils isolated from blood of cadavers was higher in comparison with the same parameter in neutrophils isolated from survivor's blood. The significance of this observation for resuscitation and transplantology is discussed.     

A radioautographic study on RNA synthesis in aging mouse spleen after 3H-uridine labeling in vitro.

EM radioautographic study on RNA synthesis in aging mouse spleen was conducted after 3H-uridine labeling in vitro. The localization of radiolabelled precursor was used to determine the site of RNA synthesis. The site of the radiolabelled uridine uptake was localized in the haematopoietic cells, particularly in the lymphoblasts. In the labelled cells, most of the silver grains were localized in the nucleus, specifically in the euchromatin. Few cytoplasmic organelles such as the mitochondria and endoplasmic reticulum were labelled with 3H-uridine. Silver grains were also observed over the nucleoli. The labeling index was expressed as the percentage of labelled cells over the total number of cells counted. The labeling index increased from day one after birth and progressively until the 14th day. Thereafter, the labeling index decreased gradually until the 10th month. A significant difference of p less than 0.05 was noted. In all the EMRAG analyzed, it was observed that the number of silver grains per cell increased proportionally with the labeling index. The result of the quantitation of the changes in RNA synthesis correlated well with the maturational development/aging of the animal.     

RNA synthesis in embryogenesis of Drosophila virilis]

The synthesis of different RNA fractions labelled by 3H-uridine has been studied at different developmental stages in Drosophila virilis by means of thermal phenol extraction and chromatography on oligo-alphaT-cellulose. There are two periods of intensive synthesis of poly-A containing (messenger) RNA in the course of embryogenesis. The former (6--12 hrs) is the period of gastrulation and stomodeum development and the latter (15--21 hrs) is the period of intensive differentiation of larval organs. The synthesis of poly-A free RNAs predominates at the stages of blastoderm (2--5 hrs) and mesoderm segmentation and embryo shortening (12--15 hrs).     

A comparative radioautographic analysis of DNA synthesis by enterocytes of the small intestine in in vivo and in vitro experiments

A method of radioautography was used to study DNA synthesis in mice enterocytes as an index of their proliferative activity. Cells were labelled by means of incorporation [3H]thymidine in vivo and in vitro. The program was worked out to computed analysis of the reflected light intensity of obtained radioautographs and estimation of reliability data. Our experiments showed that thymidine diffused very slowly.     

Electron microscopic and radioautographic data on the synthesis of RNA and chromatin structure in cells of the rat cerebral cortex

The intensity of RNA synthesis in cells of the rat brain hemispheres (neurons, astrocytes, oligodendrogliocytes, microgliocytes) was studied by electron microscopic radioautography, and the data obtained were compared with dispersed to condensed chromatin area ratio. The correlation was found between the level of RNA synthesis and dispersed chromatin area. High chromatin dispersity in neuron and intensive NA synthesis in the extranucleolar part of the nucleus made it possible to assume the existence of depression of an especially large genome part and the variability of the proteins produced by this cell.     

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.     

Heliomycin suppression of RNA synthesis in a cell-free system]

Heliomycin inhibited in vitro the RNA-polymerase reaction catalyzed by the preparation of DNA-dependent RNA-polymerase from E. coli. The blocking effect increased with a rise in the antibiotic concentration. The inhibitory effect of heliomycin decreased, when the amount of RNA-polymerase in the system increased. Yet, it did not depend on the content of DNA and the nature of the DNA preparation. Preincubation of RNA-polymerase with DNA resulting in formation of the enzyme-matrix complex did not prevent blocking RNA synthesis by heliomycin. Suppression of the RNA-polymerase reaction did not depend on the time of the antibiotic addition to the polymerizing system. Heliomycin had a significant activity not only with respect to the bacterial RNA-polymerase, but also in the system containing the enzyme isolated from the cells of Crithidia oncopelti.     

A radioautographic study of collagen synthesis by earthworm epidermis

Secreation by the epidermis of two oligochaetes (Eisenia and Enchytraeus) was investigated radioautographically following administration of 3H-proline, 3H-tryptophan or Na2(35)SO4. Regionally epidermal columnar cells of Enchytraeus synthesize the overlying, probably collagenous, cuticle. Eisenia epidermis does not recordably synthesize the cuticle until after wounding (first eight segments removed). By two days postoperative the epidermal columnar cells of Eisenia synthesize the collagenous cuticle and, later in regeneration, the epidermis may simultaneously synthesize the different collagen of the underlying basement lamella. The epidermis of Enchytraeus, but not of Eisenia, synthesizes some sulfated material associated with the cuticle surface.