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Biomedical subjects

C A Bourgeois

Publications and source records attributed to C A Bourgeois.

At least 37 records · Page 2Linked to original sources

Evidence for the existence of a nucleolar skeleton attached to the pore complex-lamina in human fibroblasts.

This work deals with the types of nuclear skeletal structures obtained from human fibroblast nuclei isolated by different procedures. It is confirmed that, in somatic vertebrate cells, the pore complex-lamina is always observed, whereas the presence of internal nucleolar and extranucleolar residual structures depends upon the method of nuclear isolation used. Furthermore, the results reported here argue for the existence of a nucleolar skeleton different from the nucleolar matrix often observed in different cell types by other investigators. The conditions of nuclear isolation which allow us to visualize this nucleolar skeleton without any other internal residual structures are described. The attachment of the nucleolar skeleton to the lamina suggested by the present data is considered in relation to the in situ position of nucleoli near the nuclear envelope.

Cell Nucleolus↗

Detection of sugar-binding sites in the fibrillar and the granular components of the nucleolus: an experimental study in cultured mammalian cells.

The intranucleolar distribution of sugar-binding sites (i.e., lectin-like molecules) was analyzed in segregated nucleoli of actinomycin D-treated HeLa cells. The detection of sugar-binding sites was performed by incubation either of permeabilized nuclei in the presence of fluorescein-labeled neoglycoproteins or of ultrathin sections cut through in situ-fixed nuclei in the presence of gold-labeled neoglycoproteins. In the former case, the fluorescent nucleolar components were identified by comparison with the nucleolar components of similarly treated cells observed in electron microscopy. For the first time, this study reveals the presence of sugar-binding sites in both the fibrillar and the granular components of the nucleolus. In view of the data already reported on the biochemical composition of the nucleolus, some of our results led us to conclude that the nucleolar sugar-binding sites are lectin-like proteins. These proteins could be associated with preribosomes since the nucleolus is the site of both synthesis and stockage of ribosomal precursors. Some results from this study, however, show that the possibility of a relationship between some lectins and a structural component cannot be excluded.

Binding Sites↗

The nuclear skeleton and the spatial arrangement of chromosomes in the interphase nucleus of vertebrate somatic cells.

The topologic distribution of interphase chromosomes established by using various cytologic methods and data concerning the DNA-nuclear skeleton interactions in isolated nuclear fractions were reviewed and discussed. Comparison of these different data clearly showed that the position of chromosomes observed in situ is in agreement with the results obtained from isolated nuclear fractions, indicating that all DNA molecules are bound to the peripheral nuclear skeleton. Moreover, the in situ position of the rDNA near the nuclear envelope can be correlated with the existence of a nucleolar skeleton connected to the peripheral nuclear skeleton. Taking into account the discrepant results regarding the actual existence of an internal nuclear skeleton, we attempted to analyze how the various nuclear skeletal structures described in the literature can be involved in both the distribution of chromosomes and in their chromatin organization. As many questions are still unanswered, we considered the modes of investigation that seem to be the most promising.

Animals↗

New data on the in-situ position of the inactive X chromosome in the interphase nucleus of human fibroblasts.

The in situ spatial distribution of nucleolus-organizing-region (NOR) bearing chromosomes in relation to the inactive X chromosome was studied during interphase in human fibroblasts. The respective positions of these chromosomes were examined in 30 growing and 32 resting fibroblasts from reconstructed nuclei, using nucleoli and the Barr body as ultrastructural markers. Experimental values for the distance between the nucleoli and the Barr body were estimated by their coefficient of closeness and compared to the uniform distribution. The following results were obtained: the distribution patterns for the two populations of nuclei were similar, the distribution of the NOR-bearing chromosomes in relation to the inactive X chromosome varied and differed significantly from a uniform distribution, and in many cases the Barr body was observed to be in a juxta-nucleolar position. The internal distribution revealed by this study is compared with the data in the literature, especially with the conflicting data obtained by other methods used to determine the interphase arrangement of chromosomes. The relationship between interphase and metaphase arrangements such as can be deduced with these methods, is discussed in relation to the mechanisms of the formation of metaphase plates or chromatid translocations.

Cell Nucleolus↗

Rotation of the cell nucleus in living cells: a quantitative analysis.

Nuclear rotation is observed in a variety of cell types. However, few quantitative analyses are reported and the significance of this phenomenon is still unclear. To investigate this type of nuclear movement, we performed a quantitative analysis in mouse L-929 fibroblasts, a cell line chosen since it displays a high nuclear rotational activity. Analyses were performed using time-lapse microcinematography. The relationship between nuclear rotation and other cellular phenomena such as the cell cycle and locomotion were studied. Then, we investigated the rotation in a population of sister cells to study whether it is genetically determined. Finally, we performed a qualitative analysis of nuclear rotation in different cultured cell lines. Results show that nuclear rotations preferentially occur during the phases of the cell cycle which surround mitosis.

Animals↗

In situ arrangement of nor-bearing chromosomes in the interphase nucleus of Aotus trivirgatus.

The spatial organization of the two nucleolus-organizing region (NOR)-bearing chromosomes during interphase was studied in Aotus trivirgatus fibroblasts, using nucleoli as ultrastructural markers. Their distribution was examined by measuring the distances between them in 30 reconstructed nuclei, and comparing these experimental values with the theoretical ones obtained by simulation. Results were as follows: (1) the nucleoli are arranged in a polarized manner inside the nucleus; (2) the nucleoli are tightly bound to the nuclear envelope at two opposites sites; (3) the distance between the two nucleoli is variable, and is shorter than it would be if the two nucleoli were distributed at random. These findings indicate that the NOR-bearing chromosomes are fixed at the nuclear envelope in two opposite areas. They are also consistent with the hypothesis that each chromosome occupies a separate domain inside the nucleus. They can be interpreted according to the model in which chromosome arrangement within the interphase nucleus is based on the separation of the diploid complement into two independent haploid sets.

Animals↗

Evidence showing that the nucleolus-envelope region is located on the nor-bearing chromosomes in Aotus trivirgatus.

We investigated whether the existence of the Nucleolus-Envelope Region observed in dividing animal cells is related to the Nucleolus Organizer Region or to other chromosome segments of the NOR-bearing chromosomes, since NORs are usually adjacent to the chromosome segments attached to the nuclear envelope such as the centromere, telomere or heterochromatin region. We used Aotus Trivirgatus fibroblasts whose karyotype is characterized by a single pair of NORs located on the long arm of the third pair of chromosomes, far from the centromere, the telomere and any obvious heterochromatin region. All the nucleoli were seen to be clearly associated with the nuclear envelope but separated from it by the outermost layer of chromatin. These results further support the hypothesis that the NOR is a site of attachment of the chromatin to the nuclear envelope by means of the Nucleolus-Envelope Region. They show that genetically active chromatin is also attached to the nuclear envelope. A model of the arrangement of chromatin during interphase is proposed which provides a functional interpretation of the currently available data.

Animals↗

Effect of low dose rate ionizing radiation on the division potential of cells in vitro. VIII. Cytogenetic analysis of human fibroblasts.

Human embryonic and adult cells were irradiated with fractionated doses of low dose rate ionizing radiation starting early during their lifespan. Adult cells were found to be more sensitive than fetal cells to ionizing radiation in terms of the number of cells produced during the lifespan of the control and the irradiated cultures. Phase-III adult control cells had fewer chromosomal aberrations than phase-III embryonic control cells. After irradiation there was an increase in chromosomal aberrations in adult cells but no increase in embryonic cells beyond those found in the control cultures. It is suggested that cells that have a higher potential for chromosomal rearrangements survive better after low dose rate ionizing radiation.

Adult↗

Analysis of the spatial organization of the cell: a statistical method for revealing the non-random location of an organelle.

A method is proposed for testing the randomness of the location of an organelle within a given area of a cell section. The approach chosen is to analyse the distance between this organelle and a specific point considered as a point of reference. The method consists of converting this distance into the ratio of one given area to another and comparing the statistical distribution of the converted values to the uniform distribution. This method has the advantage of being valid in the absence of any restrictive assumptions concerning the heterogeneity in size and/or shape of the collection of sections sampled. Detailed examples are given to illustrate the practical use of the method, and its possible extensions are discussed.

Cell Nucleolus↗

Ultrastructural localization of Ag-NOR stained proteins in the nucleolus during the cell cycle and in other nucleolar structures.

EM investigation of Ag-AS-NOR staining after short glutaraldehyde prefixation followed by Carnoy fixation maintained good ultrastructural preservation and reactive selectivity. This enables exact localization of silver deposits both in the fibrillar centers of typical or segregated nucleoli during interphase, and in chromosome NORs during mitosis. These results argue in favour of the possibility that fibrillar centers are the interphasic counterpart of chromosome NORs. Special structures such as nucleolar blobs and remnants usually considered to be of nucleolar origin, were also stained. - These findings seem to indicate a relationship between the distribution of the silver-stained proteins, the arrangement of the nucleolar structures and the degree of nucleolar activity resulting from the experimental conditions. These results are of interest at the time when the concept of the nucleolar matrix is gradually emerging.

Animals↗

Fanconi anemia leading to acute myelomonocytic leukemia: cytogenetic studies.

A 6-year-old boy with Fanconi anemia, developed acute myelomonocytic leukemia with marker chromosomes from an interchange in the malignant clone in the bone marrow. The possible aetiological role of therapy, the morphological characteristics and chromosome aberrations of the bone marrow cell line are discussed in relation to previous reported cases of Fanconi anemia in which acute leukemia has supervened.

Anemia, Aplastic↗