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[Analysis of the polyploidization kinetics of the parenchymal cells in the rat liver].

Methodological approaches to kinetics of cell polyploidization in the rat liver parenchyma are discussed. Different ways of hepatocyte polyploidization in the course of postnatal liver growth have been assessed. The intensities of hepatocyte transitions from one ploidy class to another were determined. On the basis of literary experimental data the following is summarized: With the increase in the animal age, there is a decrease in hepatocyte transition from one ploidy class to and ther; in young animals the intensity of formation of tetraploid hepatocytes through the stage of binuclear cells (2c----2c X 2----4c) is 0.39-0.55 within two weeks, the intensity of direct transitions (2c----4c) being 0.00-0.19 within the same time. The intensity of entering to DNA synthesis is reduced with the increase in hepatocyte ploidy levels; in this case the coefficient of the reducing of mitotic activity is calculated as 0.10-0.22, and 0.01-0.05 for 4c- and 8c-hepatocytes, resp. The factors stimulating proliferation in the liver increase the intensity of the direct cell transition (2c----4c) by several times which can exceed the intensity of transition through the binuclear cell stage.

Aging↗

Growth arrest and polyploidization induced by metahalone microtubule inhibitors on rat glioma cells in culture.

Two pyrimidine analogs (metahalones) NY 3163, NY 3170 have been tested for their effects on the growth of rat glioma cells in monolayer and in spheroid culture. Both substances have earlier been found to inhibit the microtubule system in malignant cells. In glioma cells, arrest of mitosis was accompanied by repeated cycles of DNA synthesis, leading to different levels of polyploidization up to 16 and 32 ploid cells. The effect was not reversible through a culture period of 4 days. Reduced ability of directional migration of cells on a plastic surface was seen for 16 days after exposure. The reaction to microtubule inhibitors seems to differ depending on the type of tumour cells, where some malignant cells have earlier been reported to escape mitotic arrest and proceed to the next G1 phase, in contrast to the present glioma cells which undergo polyploidization.

Animals↗

Cell kinetics of mouse urinary bladder epithelium. III. A histologic and ultrastructural study of bladder epithelium during regeneration after a single dose of cyclophosphamide, with special reference to the mechanism by which polyploid cells are formed.

In order to see whether the polyploid cells lining the mouse urinary bladder are formed by nuclear fusion, such epithelium was studied under the light and electron microscope forty-eight hours after an injection of cyclophosphamide when the bladder epithelium regenerates with rapid formation of many diploid, tetraploid and octoploid cells. The probability of seing fusion, if it occurs, ought then to be high. Serial sections of many specimens from four mice revealed no signs of fusion. Thus we found no support for the theory that polyploid cells are formed by nuclear fusion.

Animals↗

Induction of polyploid nuclei in the plasmodium of Physarum polycephalum by platinum antitumor compounds.

The intranuclear mitosis of the plasmodial nuclei of myxomycetes permits the observation of defects in chromosomal repartition which would probably be lethal in other eukaryotic cells with open mitosis. We found that antitumoral platinum-amine compounds perturbed late mitotic events and induced the formation of giant nuclei which were polyploid in plasmodia of Physarum polycephalum. Using 26 platinum-amine complexes, we have shown that all antitumoral compounds induced the formation of polyploid nuclei for drug concentrations at least three times lower than the amount necessary to block the overall plasmodial growth, whereas platinum compounds without antitumor activity did not behave this way. DNA replication appeared to be quantitatively normal during formation of giant nuclei by antitumoral compounds. These observations suggest that platinum-amine compounds exert their antitumor activity by interfering with mitosis rather than by a gross inhibition of DNA synthesis.

Cell Nucleolus↗

[Effect of polyploidization on the anchorage-independent multiplication of transformed cells].

Three nearhexaploid sublines were obtained from hypotriploid mouse L cells by means of colcemid treatment. When cultivated on solid substratum, all of them did not differ from the parental line either in doubling time or in cloning efficiency. The ability of polyploid cell variants to be initiated for proliferation in a semi-solid medium was equal to that of hypotriploid cells, while the average diameter of colonies formed by hexaploid cells in methyl cellulose turned out to be significantly smaller than the size of colonies of parental cells. The inhibition of growth in the semi-solid medium may reflect partial normalization of the transformed phenotype of polyploid L cells.

Animals↗

[Polyploidization mechanisms of the cardiac myocytes in the mouse].

Ventricular cardiomyocytes of 5-6 day-old mice are mostly polyploid. At this time 60% of the cardiomyocytes become binuclear and another 10% mononuclear polyploids. The binuclear cardiomyocytes emerge as a result of acytokinetic mitosis, while mononuclear tetraploids as a result of the cessation of the mitosis at the initial stages.

Animals↗

[Polyploidization of hepatocytes under different regimens of rat liver exposed to CCl4].

Polyploidization of hepatocytes in albino rats subjected to continuous and intermittent exposure to CCl4 has been studied karyometrically. It has been shown that polyploidization as a response to the exposure of chlorderivatives of hydrocarbons is determined by hepatotoxin toxicity related to the number of chlorine atoms in the molecule and to the character of the exposure involving route, dose and regimen of exposure.

Animals↗

Polyploidization by means of endoduplication in a human breast cancer cell line.

Near DNA diploid human adult solid tumors are often associated with certain near-tetraploid cells. In an established human breast cancer cell line, Hs578T, with a DNA index in the hyperdiploid region, polyploid cells appeared during exponential growth. Among clones generated from single cells and analyzed by the video time lapse technique, an intraclonal interdivision time (IDT) heterogeneity is presented that renders endoduplication a plausible explanation for the generation of the polyploid cells observed. This conclusion, drawn from our IDT analysis, is supported by curves drawn from counting grain-positive cells during continuous labeling with [3H]-thymidine. Our results are compared with a parallel analysis of the aneuploid human breast cancer cell line MDA-231, generating intraclonal IDT heterogeneity, due mainly to the mitotic instability of that line, as we reported previously.

Breast Neoplasms↗

[Polyploid mitoses in mink trophoblast cells].

Mitotic figures in the mink placental trophoblasts have been observed under the light microscope using actions and air-dried preparations. The tetra- and octaploid metaphase chromosome spreads were found on Giemsa-stained air-dried preparations. A high percentage (up to 80%) of abnormal metaphases, including k-mitoses as well as a portion of restitution anaphases, was revealed on sections of the placental trophoblast suggesting a possible block of mitosis at the meta- and anaphase. Therefore, it is very likely that genome multiplication in a portion of placental trophoblast cells in mink involves block of mitoses at meta- and anaphase followed by restitution. The chromosomal arrangement on metaphase spreads in part of cells showed a degree of separation of the whole di- and tetraploid chromosome sets within tetraploid and octaploid chromosome plates. Several spreads exhibited some allocycly of diploid chromosome sets inside the polyploid metaphases. It is not inconceivable that such an arrangement may reflect some autonomy of low-ploidy chromosome sets within the polyploid trophoblast cells in mink.

Animals↗

Polyploid accumulation associated with hyper-phosphorylation of histone H1 in hepatocytes of LEC mutant rat.

In this study, we analyzed effects of copper toxicity on the process of mitosis by examining polyploid accumulation and histone H1 hyper-phosphorylation, which result from defects in mitotic completion, in regenerated LEC rat liver. The proportion of polyploids with 8c nuclear DNA content in LEC rats was significantly higher than that in normal rats (P < 0.05). The degree of histone H1 phosphorylation was about 2.5-fold higher in LEC rat than normal rat. These results suggest that the accumulated copper in LEC mutant rat causes the block of mitotic completion in cell cycle.

Animals↗

[Hepatocyte polyploidization in chronic liver diseases in man].

Levels of hepatocyte ploidy were investigated in patients with chronic viral hepatitis, alcohol hepatitis, and liver cirrhosis. It was shown that in spite of some decrease in the share of mononucleate diploid hepatocytes these cells equally make the main part of hepatocyte population (about 80% on the average) both during chronic liver diseases and in the norm. In chronic hepatitis and cirrhosis processes of polyploidization in the human liver, unlike those in livers of laboratory animals (rats and mice), are accomplished with much lower rates and characterized by accumulation of binucleate diploid cells. These processes differ from those occurring under ageing. Reparative regeneration of human liver under chronic hepatitis and cirrhosis is accomplished on the whole by proliferation of mononucleate diploid hepatocytes but processes of polyploidization in the liver of some patients can play a significant role in reparative liver growth.

Adolescent↗

[Genome multiplication mechanisms in the development of albumen gland polyploid cells of Succinea lauta (Gastropoda:Pulmonata). VI. Ultrastructural research of endomitotic structure].

This ultrastructural study of endomitosis in polyploid albumen gland cells of succineid snail. S. lauta was made to extend the previous research (Anisimov, 1997). The investigation was prosecuted on ultrathin sections by means of transmission electron microscopy. The insertion of 3H-thymidine and 3H-uridine into the nuclei and the light microscopy control of nuclear ploidy allowed to identify glands on early and intensive polyploidization stages and to ignore pseudoendomitotic terminally differentiated cells. Considering ultrastructural qualities and information about the cell population kinetics enabled us to discriminate between predifferentiation (2-4c), protodifferentiation (4-8c) and promoted differentiation (8-16c) cell stages. The endomitotic cycle was observed synchronously with the development of cytoplasmatic structures and secretion production in differentiating cells of ploidy levels 4-8-16c. According to morphological condition of the chromatin and chromosomes, G1-, S- and G2-interphase periods can be determined, such as successive stages of endomitosis (endopro-, meta-, ana-, and telophase). The following main features of endomitosis are accentuated: the normal (mitotic) chromosome cycle with total compaction in endometaphase and differential decompaction in interphase; the absence of the mitotic spindle; integrity of the nuclear envelope, nucleolus and contacts between them and chromosomes; part of chromosomes split nonsimultaneously and incompletely in the endoanaphase, or endomitosis with diplochromosomes rarely occurs. Chromosome nondisjunction is a temporary process that does not lead to a permanent chromosome endoreduplication (polyteny). It is emphasized that the normal chromosome cycle is combined with a complete block of cytoplasmic structures. Possible reasons of mitotic cycle reduction during the transition of differentiating cells to endomitotic polyploidy are discussed.

Animals↗

[Genome multiplication mechanisms in the development of albumen gland polyploid cell of Succinea lauta (Gastropoda:Pulmonata). VIII. Pseudomitosis in the terminally differentiated cells].

Using a mature albumen gland of Succinea lauta, a study was made of the chromatin structure and RNA synthesis activity in polyploid nuclei (16c-32c-64c) of terminally differentiated cells, by means of electron microscopy and 3H-uridine radioautography of squash preparations. No 3H-thymidine incorporation in these cells was observed, which suggested the absence of DNA synthesis and endomitotic cycles at the examined functional stage of gland development. Some different stages of secretory cells could be distinguished: a resting period of the terminally differentiated cells, with the least 3H-uridine incorporation; a phase of intense heterosynthetic transcription, with the highest 3H-uridine incorporation; intervals in daily cycles; nuclear degeneration in ageing cells. At all these states the nuclei demonstrated some morphological features of endomitosis. This may be possible, because in the course of terminal cell differentiation the main chromatin mass is compacting like chromatids or chromocentres with thin euchromatinous filaments, which can be covered with perichromatinous granules. Since the true endomitosis is deliberately excluded at this stage, the above described morphological state of polyploid nuclei is regarded as postendomitotic and is designated as pseudoendomitosis. The view of the existence of "two fundamentally different types of endomitosis", one of which is "endomitosis in general", and the other "shows up as stationary" (Therman et al., 1983), is accepted as groundless.

Animals↗

Human herpesvirus 7 infection induces profound cell cycle perturbations coupled to disregulation of cdc2 and cyclin B and polyploidization of CD4(+) T cells.

Human herpesvirus 7 (HHV-7) infection of both primary CD4(+) T lymphocytes and SupT1 lymphoblastoid T-cell line induced a progressive accumulation of cells exibiting a gap 2/mitosis (G2/M) and polyploid content coupled to an increased cell size. The expression of both cyclin-dependent kinase cdc2 and cyclin B was increased in HHV-7-infected cells with respect to the uninfected ones. Moreover, the simultaneous flow cytometric analysis of cyclin B and DNA content showed that cyclin B expression was not only increased but also unscheduled with respect to its usual cell cycle pattern. However, the levels of kinase activity associated to cdc2 were decreased in HHV-7-infected cells with respect to uninfected cultures. To elucidate the origin of the enlarged HHV-7-infected cells, extensive electron and confocal microscopy analyses were performed. Membrane fusion events associated to cytoplasmic bridges, which characterize the formation of syncytia, were never observed. On the other hand, analysis of serial sections of the same cells strongly suggested that enlarged HHV-7-infected cells contained a single polylobated nucleus. This was confirmed by flow cytometry analysis performed on nuclei isolated from HHV-7-infected cells, which showed multiple peaks with a DNA content >4n. Taken together, these data indicate that giant cells, which represent the hallmark of in vitro HHV-7 infection, arise from single CD4(+) T cells undergoing a process of polyploidization.

CD4-Positive T-Lymphocytes↗

Rapid structural and epigenetic changes in polyploid and aneuploid genomes.

Recent work with plants has demonstrated that genome instability can be triggered by a change in chromosome number arising from either whole genome duplications (polyploidy) or loss/gain of individual chromosomes (aneuploidy). This genome instability is manifested as rapid structural and epigenetic alterations that can occur somatically or meiotically within a few generations after heteroploid formation. The intrinsic instability of newly formed polyploid and aneuploid genomes has relevance for genome evolution and human carcinogenesis, and points toward recombinational and epigenetic mechanisms that sense and respond to chromosome numerical changes.

Aneuploidy↗

Fra(X) prenatal diagnosis: are endoreduplicated and polyploid cells useful diagnostic criteria?

Cytogenetic and molecular protocols for prenatal ascertainment of the fragile X syndrome and the associated fragile site at Xq27.3 are relatively reliable. Any new diagnostic method which becomes available still elicits much interest. Kimchi-Sarfaty et al. [1991] reported an increase in frequency of endoreduplication and polyploidy in fra(X) lymphoblasts and amniocytes when cultured with methotrexate (MTX) or fluorodeoxyuridine. Recently we analyzed the endoreduplication/polyploidy system using amniotic fluid, chorionic villus, and fibroblasts from fra(X) positive abortus cell cultures and from control samples. We observed no increased expression of endoreduplicated or polyploid cells in fra(X) positive amniocytes after exposure to MTX. The data presented here clearly dispute the value of endoreduplication/polyploid scoring as a diagnostic aid in prenatal fra(X) analysis.

Chromosome Aberrations↗

Specific nuclear elimination in polyploid plasmodia of the slime mold Physarum polycephalum.

In growing plasmodia of the myxomycete Physarum polycephalum (G2-phase), three distinct classes of nuclei with a relative DNA content of 1x, 2x, and 4x are observed in the presumed haploid strain CL. The 2x and 4x species comprise up to 35% and 5% of the nuclei. Quantitative cytofluorometric studies of nuclei isolated in either G2- or S-phase or after FUDR treatment (G1 arrest) show that the three nuclear populations undergo a synchronous mitotic cycle and that the relative DNA content of the nuclear fractions in G-2 phase reflects the 2c, 4c, and 8c state. The heterogeneity of the nuclear population does, however, seem to be restricted to the growth phase. During a starvation period of 4 days that always preceeds sporulation (and also meiosis), the 4c nuclear population is reduced to 7%, 8c nuclei are no longer detected. These results suggest that a mechanism exists in Physarum for the selective detection and elimination of polyploid nuclei.

Cell Nucleus↗

Control of DNA synthesis in polyploid mammalian cells.

To test the hypothesis that the duration of DNA synthesis is an inverse function of nuclear size or DNA content, the S phase was calculated from PLM analysis for pseudodiploid, tetraploid, and octaploid lines of Chinese hamster cells growing as a monolayer or in suspension. S phase times were found not to be significantly different between polyploid lines and the diploid lines from which they were derived, regardless of the conformation of the nucleus. There is no evidence, therefore, that would implicate the nuclear membrane, or nuclear surface area/volume relationships, in the control of DNA synthesis.

Animals↗