Modulation of lymphocyte nuclear matrix organization <i>in vivo</i> by 5,6‐dichloro‐1‐β‐D‐ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study

Chaly, Nathalie; Cadrin, Monique; Kaplan, Joseph; Brown, David L.

doi:10.1111/j.1768-322x.1988.tb00736.x

Cited by 17 publications

(4 citation statements)

References 57 publications

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“…The results are also consistent with observations on nuclear reorganization in mitogen-stimulated lymphocytes, in which increases in nuclear volume, enlargement of nucleoli and the development of extensive interchromatin regions are paralleled by increases in the number and density of nuclear pores (Maul et al 1972), the influx and quantity of non-histone nuclear proteins (Setterfield et al 1983(Setterfield et al , 1985, and the immunofluorescence labeling pattern and intensity of antibody PI1 (Setterfield et al 1985;Chaly et al 1988).…”

Section: Discussionsupporting

confidence: 79%

“…This pathway is not necessarily restricted to integral components of chromatin such as histones, but may also be followed by other nuclear proteins that bind transiently to the chromosomes prior to nuclear envelope breakdown. Examples of nuclear proteins that are" piggy-back" on chromosomes include a number of nucleolar components, such as RNA polymerase I (Scheer and Rose 1984;Matsui and Sandberg 1985), protein C23 or nucleolin (Lischwe et al 1981), fibrillarin , N038 or B23 (Ochs et al 1983;Schmidt-Zachmann et al 1987), ribocharin (Hiigle et al 1985b) and preribosomal particles (Hiigle et al 1985a) as well as DNA topoisomerases I (Guldner et al 1986) and 11 (Earnshaw and Heck 1985), perichromin (McKeon et al 1984), and peripherin (formerly P1) (Chaly et al 1984(Chaly et al , 1988.…”

Section: Discussionmentioning

confidence: 99%

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Functional role of newly formed pore complexes in postmitotic nuclear reorganization

et al. 1989

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Abstract. Many nuclear proteins are released into the cytoplasm at prometaphase and are transported back into the daughter nuclei at the end of mitosis. To determine the role of this reentry in nuclear remodelling during early interphase, we experimentally manipulated nuclear protein uptake in dividing cells. Recently we and others have shown that signal-dependent, pore complex-mediated uptake of nuclear protein is blocked in living cells on microinjection of the lectin wheat germ agglutinin (WGA), or of antibodies such as PI1 that are directed against WGA-binding pore complex glycoproteins. In the present study, we microinjected mitotic PtK z cells with WGA or antibody PIt and followed nuclear reorganization of the daughter cells by immunofluorescence and electron microscopy. The inhibitory effect on nuclear protein uptake was monitored by co-injection of the karyophilic protein nucleoplasmin. When injected by itself early in mitosis, nucleoplasmin became sequestered into the daughter nuclei as they entered telophase. In contrast, nucleoplasmin was excluded from the daughter nuclei in the presence of WGA or antibody PI1 . Although PtK z cells with blocked nuclear protein uptake completed cytokinesis, their nuclei showed a telophaselike organization characterized by highly condensed chromatin surrounded by a nuclear envelope containing a few pore complexes. These findings suggest that pore complexes become functional as early as telophase, in close coincidence with nuclear envelope reformation. They further indicate that the extensive structural rearrangement of the nucleus during the telophase-G1 transition is dependent on the influx of karyophilic proteins from the cytoplasm through the pore complexes, and is not due solely to chromosome-associated components.

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Section: Discussionsupporting

confidence: 79%

Section: Discussionmentioning

confidence: 99%

Functional role of newly formed pore complexes in postmitotic nuclear reorganization

et al. 1989

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“…In through-focus observations, much of the labelling appeared to be associated with regions of aggregated, condensed chromatin (Figure 3a & a', 4a & a'). Lymphocyte response to mitogen activation is asynchronous, so that 48 h-stimulated populations contain small unstimulated cells, large fully stimulated cells, and cells at intermediate stages of activation ( Figure 3b') (Setterfield et al 1983, Chaly et al 1988. In these samples (Figure 3b & b'), the anti-Topo II:~ labelling was unchanged in unstimulated cells, and the labelling intensity increased in parallel with the extent of stimulation.…”

Section: Topo II Organization During the Hela Cell Cyclementioning

confidence: 85%

Organization of DNA topoisomerase II isotypes during the cell cycle of human lymphocytes and HeLa cells

et al. 1996

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We have monitored the organization of DNA topoisomerase II (Topo II) in relation to chromatin disaggregation during mitogen stimulation of lymphocytes and to the mitotic chromosome condensation cycle by immunofluorescence microscopy with isozyme-specific antibodies. Labelling for both Topo II alpha and Topo II beta was diffusely nucleoplasmic and non-nucleolar in resting lymphocytes and the pattern changed little during stimulation. Topo II alpha labelling intensity increased in parallel with the extent of cell stimulation, but a fraction of fully stimulated cells was labelled very brightly. Topo II beta labelling intensity was also greater in stimulated cells, but all partially and fully stimulated cells were labelled at the same, higher, intensity. In addition, anti-Topo II beta detected a few small spots within nucleoli of stimulated cells that coincided with regions containing fibrillarin. In lymphocytes and HeLa, chromosome association of Topo II alpha began in prophase and lasted throughout mitosis. In contrast, Topo II beta stayed nucleoplasmic in prophase, was diffusely cytoplasmic during mitosis, and was first detected post-mitotically in nuclei with decondensing chromosomes and a reformed nuclear envelope. The results are consistent with a role for Topo II alpha, but not for Topo II beta, in mitotic chromosome condensation, and indicate that the isotypes may play independent roles in the reorganization of chromatin structure during lymphocyte mitogenic activation.

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“…The nuclei are small (ca. 5 µm diameter), with large masses of condensed chromatin, small nucleoli, and very little nucleoplasm and nuclear matrix (Setterfield et al 1985;Bladon et al 1988;Chaly et al 1988). Mouse and rat spleen and thymus lymphocytes have been reported to undergo apoptosis spontaneously, with 20-50% of the population succumbing within 16 h in culture (Migliorati et al 1992;Illera et al 1993;Perandones et al 1993;Zhang et al 1995).…”

Section: Introductionmentioning

confidence: 99%

Unique behaviour of NuMA during heat-induced apoptosis of lymphocytes

Sodja

Walker²,

Brown³

et al. 1997

Biochem. Cell Biol.

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Nuclear collapse is a key feature of apoptosis, reflecting the DNA and protein fragmentation observed biochemically. We have compared nuclear events during spontaneous and heat-induced (42 degrees C for 30 min) apoptosis at the level of individual cells, monitoring overall chromatin organization by staining with 4,6'-diamidino-2-phenylindole (DAPI), DNA cleavage by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL), and the nuclear antigens NuMA, PI2, and lamin B by immunofluorescence microscopy. The data were correlated with analyses at the population level by flow cytometry and immunoblotting. We show that heat treatment doubles the level of apoptotic splenocytes with respect to the spontaneous apoptosis in controls within 6 honThe organization and disassembly of nuclear envelope antigens is identical during spontaneous and heat-induced apoptosis and proceeds in a temporally and spatially ordered manner relative to DNA fragmentation and chromatin collapse. In contrast, NuMA organization is affected by heat treatment, with about half of the cells containing many bright spots in addition to the usual diffuse labelling. The spots were still visible in many fully collapsed apoptotic nuclei. Further, this novel reorganization of NuMA and the hyperinduction of apoptosis by heat are correlated, suggesting that lymphocytes with reorganized NuMA are destined to undergo apoptosis. The data also indicate that DNA fragmentation precedes extensive remodelling of the nucleoplasm in these cells, and that chromatin begins to collapse before disassembly of nuclear envelope components.

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Modulation of lymphocyte nuclear matrix organization in vivo by 5,6‐dichloro‐1‐β‐D‐ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study

Cited by 17 publications

References 57 publications

Functional role of newly formed pore complexes in postmitotic nuclear reorganization

Functional role of newly formed pore complexes in postmitotic nuclear reorganization

Organization of DNA topoisomerase II isotypes during the cell cycle of human lymphocytes and HeLa cells

Unique behaviour of NuMA during heat-induced apoptosis of lymphocytes

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