Ultrastructural nucleolar alterations induced by an ametantrone — poly r(A-U) complex

Jamison, James M.; Gilloteaux, Jacques; Thiry, Marc; Authelet, Michèle; Goessens, Guy; Summers, Jack L.

doi:10.1016/s0040-8166(98)80061-x

Cited by 3 publications

(2 citation statements)

References 29 publications

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“…Subsequently, the cells were dehydrated through graded ethanol solutions and then embedded in epoxy resin. Ultra-thin sections of the various blocks were either collected in platinum rings (diameter 5 4 mm) formed by a platinum wire (diameter 5 0.1 mm) (SA Johnson Matthey; Brussels, Belgium) and stored on distilled water until used or mounted on colloidal-coated nickel grids (Thiry et al 1997;Jamison et al 1998). …”

Section: Preparation For Transmission Electron Microscopymentioning

confidence: 99%

Nucleolar Changes and Fibrillarin Redistribution Following Apatone Treatment of Human Bladder Carcinoma Cells

Jamison

Gilloteaux²,

Perlaky

et al. 2010

J Histochem Cytochem.

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(KS) S U M M A R Y Ascorbate and menadione (Apatone) in a ratio of 100:1 kills tumor cells by autoschizis. In this study, vitamin-induced changes in nucleolar structure were evaluated as markers of autoschizis. Human bladder carcinoma (T24) cells were overlain with vitamins or with culture medium. Supernatants were removed at 1-hr intervals from 1 to 4 hr, and the cells were washed with PBS and prepared for assay. Apatone produced marked alterations in nucleolar structure including redistribution of nucleolar components, formation of ringshaped nucleoli, condensation and increase of the proportion of perinucleolar chromatin, and the enlargement of nucleolar fibrillar centers. Immunogold labeling of the nucleolar rRNA revealed a granular localization in treated and sham-treated cells, and immunogold labeling of the rDNA revealed a shift from the fibrillar centers to the condensed perinucleolar chromatin. Fibrillarin staining shifted from the fibrillar centers and adjacent regions to a more homogeneous staining of the entire nucleolus and was consistent with the percentage of autoschizic cells detected by flow cytometry. Because autoschizis entails sequential reactivation of DNase I and DNase II, and because the fibrillarin redistribution following DNase I and Apatone treatment is identical, it appears that the nucleolar and fibrillarin changes are markers of autoschizis. (J Histochem Cytochem 58:635-651, 2010)

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Section: Preparation For Transmission Electron Microscopymentioning

confidence: 99%

Nucleolar Changes and Fibrillarin Redistribution Following Apatone Treatment of Human Bladder Carcinoma Cells

Jamison

Gilloteaux²,

Perlaky

et al. 2010

J Histochem Cytochem.

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“…who showed the tissue-dependent nature of the event 11 18 – 20 and has been linked to the process of translocation and subsequent accumulation of members of the Hsp70 family in the nucleus as a result of stress 21 , 22 . All the members of this family recognized by the anti-Hsp70 monoclonal mouse antibody are made up of isoforms.…”

Section: Discussionmentioning

confidence: 99%

Skeletal Muscle for Endomyocardial Biopsy: Comparable Stress Response in Doxorubicin Cardio-myopathy

2009

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In the present study, we compared the cell damage response in skeletal and cardiac muscle tissue when exposed to doxorubicin. This was carried out by means of a less invasive informative substitute to endomyocardiac biopsy based on Hsp70 immunodetection and a subcellular analysis of the nucleolus. Male Sprague Dawley rats (62 g body weight) were randomly distributed into 3 group, the control and doxorubicin I and doxorubicin II groups, in which 15 and 25 mg/kg body weight of doxorubicin (0.1 ml, i.v.) was administered, respectively. After 15, 30, 45 and 60 minutes, portions of the left and right ventricle wall and interventricle wall, together with skeletal muscle from the posterior and anterior member, were prepared for Hsp70 immunodetection by Western blot analysis and ultrastructural study using the thin cut technique. Differential cell response between the control and treated groups was observed in Hsp70 immunodetection and at the subcellular level. In the control group, the Hsp70 recognition levels and typical normal nucleolar morphology were similar, while the treated groups showed variable-dependent Hsp70 recognition and segregation of nucleolar components, forming ring-like figures of a variable-independent nature. Comparison of cardiac and skeletal muscle tissue cell response to doxorubicin toxic aggression revealed parallelism in terms of Hsp70 accumulation in certain regions of both tissues (15 mg/kg body weight of doxorubicin), which suggests that replacing endomyocardiac biopsy analysis with skeletal muscle analysis may be a safe option.

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Quantitative real-time analysis of nucleolar stress by coherent phase microscopy

Tychinsky¹,

Kretushev²,

Klemyashov³

et al. 2008

J. Biomed. Opt.

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We develop a method of coherent phase microscopy (CPM) for direct visualization of nonfixed, nonstained mammalian cells (both cultured cells and freshly isolated tumor biopsies) followed by computer-assisted data analysis. The major purpose of CPM is to evaluate the refractive properties of optically dense intracellular structures such as the nucleus and the nucleoli. In particular, we focus on quantitative real-time analysis of the nucleolar dynamics using phase thickness as an equivalent of optical path difference for optically nonhomogenous biological objects. Pharmacological inhibition of gene transcription leads to a dramatic decrease of the phase thickness of the nucleoli within the initial minutes of cell exposure. Furthermore, the acute depletion of intracellular ATP pool, depolymerization of microtubules and inhibition of DNA replication resulted in a rapid decrease of the nucleolar phase thickness. These optical effects were paralleled by segregation of nucleolar components as documented by electron microscopy. Thus, CPM detects early changes of nucleolar dynamics, in particular, the nucleolar segregation as part of general cellular response to cytotoxic stress, regardless of whether the nucleolus is or is not the primary target of the toxin. CPM is applicable for monitoring and quantitative analysis of the "nucleolar stress" in living mammalian cells.

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Ultrastructural nucleolar alterations induced by an ametantrone — poly r(A-U) complex

Cited by 3 publications

References 29 publications

Nucleolar Changes and Fibrillarin Redistribution Following Apatone Treatment of Human Bladder Carcinoma Cells

Nucleolar Changes and Fibrillarin Redistribution Following Apatone Treatment of Human Bladder Carcinoma Cells

Skeletal Muscle for Endomyocardial Biopsy: Comparable Stress Response in Doxorubicin Cardio-myopathy

Quantitative real-time analysis of nucleolar stress by coherent phase microscopy

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