Evolution of some biochemical characteristics of the intestinal mucosa during the first postnatal weeks in C57 mice. Effects of thyroxine and putrescine.

In C57 mice, during the third week after birth, there is a rapid conversion of the intestinal epithelium from fetal to mature adult status. Some characteristics (lactase, maltase, diamine oxidase, and sucrase activity; putrescine, spermine and spermidine concentrations; mitotic index) have been analyzed for the proximal, middle, and distal parts of the intestine in mice of different ages: 8, 15, 19, and 60 days. The most important observations recorded were as follows: (a) sucrase- and maltase-specific activities as well as spermine and spermidine contents increased abruptly on the 19th postnatal day and then decreased; (b) decrease of lactase and diamine oxidase-specific activity was recorded between the 15th and 19th postnatal days, (c) later, diamine oxidase-specific activity increased, whereas putrescine content decreased in the proximal part of the intestine; and (d) mitotic index was not significantly different when estimated for the crypts on days 11 and 19. No similar variations of these biochemical parameters were observed over a period of 3 days when mice were injected with thyroxine on the 8th day or when mice received putrescine per os on the 9th day, as explained in the text. Only a slight but significant variation in sucrase- or maltase-specific activity with thyroxine and a variation of the lactase or DAO-specific activity of the distal part of the intestine with putrescine were recorded.

Ultrastructural cytochemistry of the nucleolus in rat oocytes at the end of the folliculogenesis.

Various ultrastructural changes occur during follicular growth in the rat oocyte nucleolus. The nucleolus, which has a reticulated fibrillogranular structure at the primordial and primary follicle stages, becomes entirely compact and is made up of a conspicuous and homogeneous mass at the antral follicle stage. In order to define the nature and the functions of this homogeneous mass, cytochemical methods allowing detection of nucleic acids, proteins and lipids were performed at the light microscopic and ultrastructural levels. The results obtained suggest that this nucleolar mass is probably composed of acid proteins which are not silver stained. This proteinaceous mass could be a special kind of nucleolar secretion providing material for meiotic resumption in the oocyte. Cytochemical researches now in progress should supply new information concerning the exact nature and the role of the nucleolar compact mass, which is the essential nucleolar component at the antral follicle stage and which really plays a role in the nucleolus in the first stages of embryogenesis.

Effect of prostaglandin E2 on the small intestine of indomethacin-treated rats.

In the present study, the protective effect of PGE2 on intestinal damage in indomethacin-treated adult rats was investigated. Ileal integrity was evaluated making use of different biochemical and histological parameters: activities of sucrase, maltase and diamine oxidase; concentrations of DNA, putrescine, spermidine and spermine; incorporation of 3H-thymidine into DNA; mitotic index and mucosal thickness. Results expressed per g of mucosal weight, showed that: maltase and diamine oxidase activities as well as DNA, spermidine and spermine concentrations decreased markedly in indomethacin-treated rats when compared to control rats; the decrease of maltase activity as well as DNA, spermidine and spermine concentration was less pronounced in PGE2-treated rats when compared to indomethacin-treated rats; 3H-thymidine incorporation into DNA and mitotic index values showed no significant variation in the course of different treatments; mucosal thickness increased strongly, in PGE2-protected rats. We suggest that PGE2 could protect the rat's intestinal mucosa against the effects of indomethacin through a trophic action on intestinal villi.

Behaviour of nucleolus during mitosis. A comparative ultrastructural study of various cancerous cell lines using the Ag-NOR staining procedure.

The aim of the present work was to study the distribution and the behaviour of the silver-staining nucleolar organizer region (Ag-NOR) proteins at the ultrastructural level during interphase and mitosis in five human and murine cancerous cell lines each characterized by a typical nucleolar morphology. During interphase the Ag-NOR proteins are restricted to the fibrillar centres (F.C.) and/or to the dense fibrillar component (D.F.C.). During prophase the silver-staining components come into close contact with some chromosomes and are arranged with a typical polarity: chromosome, F.C. and D.F.C. Then F.C. and D.F.C. together form roundish silver-stained structures and integrate in part within indentations at the periphery of the metaphase chromosomes. During anaphase and telophase large and small spherical silver-staining structures may be seen. They correspond respectively to the metaphase NORs and to numerous structures which appear de novo within ribonucleoprotein (RNP) material localized between the chromosomes. During late telophase the number of the small silver-staining structures decreases whereas the size of the larger ones increases. Then the interphase nucleoli recover their typical shape. These results suggest that when rRNA synthesis is impaired during mitosis the inactive NORs assume a structure and a localization which are not typical of the cell line. In contrast the F.C. and D.F.C. are probably two aspects of the NORs whose typical distribution, relative to the other nucleolar components, gives the interphasic nucleolus its characteristic morphology.

Evolution of the rat oocyte nucleolus during follicular growth.

The ultrastructural evolution of the nucleolus was followed during follicular growth by means of a silver staining procedure. The oocyte nucleolus in the primordial and primary follicles consists of strands of dense fibrillar silver-stained component and aggregates of granules which are devoid of silver grains. Small fibrillar centres are also recognized and appear to have less silver stainability. At the secondary follicle stage, a new nucleolar component appears in the reticulated oocyte nucleolus. This component is devoid of silver grains. During follicle growth, at the antral follicle stage, this new component seems to fuse and the nucleolus becomes constituted of a compact homogeneous mass which exhibits a vacuole at the end of the oocyte maturation. The results obtained suggest that this nucleolar mass is essentially made of proteins and particularly of acidic proteins.

[Nucleolar vacuoles observed in rat oocytes]. / Etude des vacuoles nucléolaires observées dans les ovocytes de rat.

Recently Merveille and al (1984) using light microscope have observed intranucleolar cavities in rat ovocytes from large antral follicles. They have showed that the frequency of these vacuoles increases when follicular growth is stimulated by gonadotropins. In this paper, the ultrastructure of the nucleolar cavities has been studied. Two types of cavities are visible in these nucleoli: 1. nucleolar "interstices" present at the periphery of the nucleoli in remnants of the granular component and of the dense fibrillar components; 2. nucleolar "vacuoles" which are located in the homogeneous substance forming the greatest part of the nucleolus. The nucleolar vacuoli generally are clear-cut and spherical. The density of their content is similar to the nucleoplasm but they don't communicate to the nucleoplasm. By means of cytochemical to detect Ag-NOR proteins (Ploton and al [1983]) and basic proteins (Sheridan and Barnett [1984]), dense fibrillar component of the nucleolus but no basic proteins may be seen in the wall of the cavity. Moreover no evidence of relation between the presence of the vacuoles and the process of follicular atresia has been found.

Effects of 2-mercapto-1-(beta-4-pyridethyl)-benzimidazole (MPB) on Ehrlich cell nucleoli: stereological analysis.

The effects of MPB, a strong inhibitor of RNA synthesis, have been analysed at the ultrastructural level by means of stereological methods. After treatment, an increase in the nucleolar volume is observed. This enlargement is due to vacuolization of the nucleoli. However, the relative volumes of the nucleolar components are modified in various directions: the volume of the granular component decreases whereas the fibrillar centres increase in size. These results are discussed in terms of relations between morphology and function of the nucleolus.

Re-evaluation of the site of transcription in Ehrlich tumour cell nucleoli.

Recent electron microscopic immunolocalization studies (Scheer and Rose, 1984) indicated that RNA polymerase I is located in, and probably confined to, the fibrillar centres of the nucleoli. This conclusion appears to be in contradiction to previous autoradiographic studies reporting that transcription of rRNA genes takes place in the dense fibrillar component. Despite the fact that rRNA transcription occurs in the dense fibrillar component of Ehrlich tumour cell nucleoli, we show in the present paper that dense fibrillar component containing rapidly labelled RNA can be visualized not only around but also inside the fibrillar centres.

[Comparative study of DNA content measured by scanning microdensitometry or by flow cytofluorometry]. / Etude comparative des teneurs en acides désoxyribonucléiques mesurées par microdensitométrie à balayage ou par cytofluorométrie de flux.

The DNA content of cultured RL12 lymphoma cells or isolated mouse liver cells was measured in the same populations, by two cytophotometrical methods: scanning and integrating microdensitometry or flow cytofluorometry. The results are compared in order to evaluate the respective advantages of the two methods which are complementary.

[Cytological study of nucleoli and nucleolar organizers during inhibition of RNA synthesis]. / Etude cytologique des nucléoles et des organisateurs nucleoaires lors d'une inhibition des synthèse d'ARN.

Adenosine induces a massive nucleolar disorganization both in chick embryo fibroblasts and Ehrlich ascitic tumour cells in which the fibrillar centre is the only persistent structure. We discuss these observations in term of RNA synthesis inhibition as fibrillar centres are the Nucleolus-Organizing Regions (NOR's).

[Effect of the spatial environment on the mitotic activity, size and protein content of Paramecium tetraurelia]. / Influence de l'environnement spatial sur l'activité mitotique, la taille, et le contenu protéinique de Paramecium tetraurelia.

Cell proliferation, cell volume, protein components of paramecia and chemical analysis of culture medium were studied in Paramecium tetraurelia cultures placed in space environment. The space flight resulted in a marked stimulation of cell proliferation, an increase of cell volume, a decrease of mean protein components and changes of electrolytes in the culture medium.

[2,4-dinitro-(pyrimidine-2-yl) thiophenol, a new drug with antimitotic properties]. / Le "dinitro-2,4 (phyrimidyl-2)-thiophénol", une nouvelle substance à propriétés antimitotiques.

A new thiopyrimidine derivative has been synthesized. It can inhibit cell multiplication in Chick embryo fibroblasts, in Mouse Ehrlich ascites tumor cells and in Rat hepatoma cells (line Rueber) cultivated in vitro.

[Demonstration by stereological methods of pre-prophase doubling of the number of ribosomes in Ehrlich tumor cells]. / Démonstration par des méthodes stéréologiques d'un doublement préprophasique du nombre de ribosomes dans des cellules tumorales d'Ehrlich.

Homogeneous populations of G1, S or G2 Ehrlich tumor cells are analysed by stereological methods at the ultrastructural level. This analysis demonstrates that the number of cytoplasmic ribosomes is doubled before the prophase.

Number of nucleoli in Ehrlich tumor cells during interphase.

Fractions of mouse Ehrlich ascites tumor cell populations with a high percentage of cell either in early or in late interphase were separated by centrifugation on ficoll gradients. Nucleoli were studied by light or electron microscopy in these cell subpopulations. It was shown that, in these cells, the number of nucleoli per nucleus does not vary significantly during interphase. This result is discussed and an anlysis of the relationships between the number and the volume of the nucleoli in these cells is present.

Selection of Ehrlich tumor cells in different phases of the cell cycle by ficoll gradient centrifugation.

Mouse Ehrlich ascites tumor cells were centrifuged at low speed on a linear Ficoll gradient. Cells from different fractions of the gradient were collected separately and analysed by cytological and cytochemical methods (DNA content, mean volume and number of cells). Nearly pure populations of cells in G1 or S were selected. A heterogeneous population containing 70% G2 and mitotic cells was also isolated. No ultrastructural alterations were detected in the cells after centrifugation. Selected G1 cells were cultured in vitro, their kinetic parameters were measured and compared with those of the original population. No difference was observed as far as the duration of the cycle of these cells is concerned.