Dissolution of cytoplasmic actin bundles and the induction of nuclear actin bundles by dimethyl sulfoxide

Sanger, Joseph W.; Gwinn, J.; Sanger, Jean M.

doi:10.1002/jez.1402130210

Cited by 26 publications

(11 citation statements)

References 15 publications

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“…They have assumed that the actin comprising the aggregates derives from a nuclear actin that is different in its properties from cytoplasmic actin (10)(11)(12). Preliminary work with Me2SO-treated PtK2 cells using fluorescent heavy meromyosin as an indicator of actin localization showed that stress fiber dissolution accompanied nuclear bundle formation (13). This suggested to us that the actin in the nuclear bundles arose from the breakdown of cytoplasmic stress fibers.…”

Section: Discussionmentioning

confidence: 99%

“…If true, this would indicate that the interphase nuclei of these cells contain a substantial amount of actin, which may play an important role in nuclear functions (10,12). On the other hand, initial studies with Me2SO and PtK2 cells (13) showed that in treated cells, bundles of actin filaments appeared in the nucleus at the same time that actin-containing stress fibers disappeared from the cytoplasm. The present results, which combine immunofluorescence with antibodies against contractile proteins and microinjection of fluorescent actin into living cells, demonstrate that the same actin that participated in the cytoplasmic stress fibers moves into the nucleus under the influence of Me2SO.…”

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confidence: 99%

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Reversible translocation of cytoplasmic actin into the nucleus caused by dimethyl sulfoxide.

Sanger¹,

Sanger²,

Kreis³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

113

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The addition of 10% dimethyl sulfoxide (Me2SO) to PtK2 and WI-38 cells caused stress fibers to disappear from the cytoplasm and numerous elongated inclusions to appear in the nucleus. When Me2SO was removed, the stress fibers reformed and the nuclear inclusions disappeared. These nuclear inclusions reacted with fluorescent heavy meromyosin, phalloidin, and actin antibody. In the electron microscope, needlelike structures were seqn-to be composed of wavy filaments that bound heavy meromyosin. Antibodies against other components of stress fibers-tropomyosin, a-actinin, and myosin-did not react with the inclusions. When fluorescently labeled actin was microinjected into living PtKX and WI-38 cells, the fluorescent actin was incorporated into stress fibers. Subsequent exposure of the same cells to Me2SO led to breakdown of the fluorescent stress fibers and the appearance of fluorescent inclusions in the nucleus. Removal of Me2SO caused reversion to the normal interphase structure. These results indicate that under the influence of Me2SO, dissolution of stress fiber releases actin in a form which allows it to diffuse into the nucleus where it then becomes organized into filamentous bundles. Evidence that actin is a constituent of the nuclei of a number of different cell types has been accumulating (1-7), although it is still not clear that its occurrence in nuclei is universal (8). The difficulty in identifying actin as an endogenous nuclear component is that the large amount of actin in the cytoplasm is a potential source of contamination of nuclear preparations. The finding by Weber and Osborn that the nucleus is surrounded by a "cage" of detergent-resistant microfilaments (9) emphasizes this problem.Recently, Fukui and Katsumaru (10-12) have shown that under the influence of 10% Me2SO, bundles of actin filaments will form in situ in nuclei of Dictyostelium, amoebae, and HeLa cells. These bundles were believed by these authors to arise from actin normally present in the nucleus. If true, this would indicate that the interphase nuclei of these cells contain a substantial amount of actin, which may play an important role in nuclear functions (10,12). On the other hand, initial studies with Me2SO and PtK2 cells (13) showed that in treated cells, bundles of actin filaments appeared in the nucleus at the same time that actin-containing stress fibers disappeared from the cytoplasm. The present results, which combine immunofluorescence with antibodies against contractile proteins and microinjection of fluorescent actin into living cells, demonstrate that the same actin that participated in the cytoplasmic stress fibers moves into the nucleus under the influence of Me2SO. There it reforms filamentous aggregates that clearly differ in composition from cytoplasmic stress fibers. This translocation of actin is rapid and fully reversible. MATERIALS AND METHODSRat kangaroo cells (PtK2) and WI-38 human fibroblasts were obtained from the American Type Culture Collection and were grown on glass cover slips in Falcon cultu...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Reversible translocation of cytoplasmic actin into the nucleus caused by dimethyl sulfoxide.

Sanger¹,

Sanger²,

Kreis³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

113

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“…To examine the role that the particular history of a given cell has in determining cell morphology, we confronted keratocytes with an acute perturbation-transient treatment with high concentrations of dimethylsulphoxide (DMSO)-which resulted in temporary lamellipodial loss and cell rounding 22 . We found that cells were able to resume movement (albeit in an arbitrary direction with respect to their orientation before DMSO treatment) and return to their original morphology and speed within minutes ( Fig.…”

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confidence: 99%

Mechanism of shape determination in motile cells

Keren

Pincus

Allen³

et al. 2008

Nature

694

816

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The shape of motile cells is determined by many dynamic processes spanning several orders of magnitude in space and time, from local polymerization of actin monomers at subsecond timescales to global, cell-scale geometry that may persist for hours. Understanding the mechanism of shape determination in cells has proved to be extremely challenging due to the numerous components involved and the complexity of their interactions. Here we harness the natural phenotypic variability in a large population of motile epithelial keratocytes from fish (Hypsophrys nicaraguensis) to reveal mechanisms of shape determination. We find that the cells inhabit a low-dimensional, highly correlated spectrum of possible functional states. We further show that a model of actin network treadmilling in an inextensible membrane bag can quantitatively recapitulate this spectrum and predict both cell shape and speed. Our model provides a simple biochemical and biophysical basis for the observed morphology and behaviour of motile cells.Cell shape emerges from the interaction of many constituent elements-notably, the cytoskeleton, the cell membrane and cellsubstrate adhesions-that have been studied in great detail at the molecular level 1-3 ; however, the mechanism by which global morphology is generated and maintained at the cellular scale is not understood. Many studies have characterized the morphological effects of perturbing various cytoskeletal and other cellular components (for example, ref. 4); yet, there have been no comprehensive efforts to try to understand cell shape from first principles. Here we address this issue in the context of motile epithelial keratocytes derived from fish skin. Fish keratocytes are among the fastest moving animal cells, and their motility machinery is characterized by extremely rapid molecular dynamics and turnover [5][6][7][8] . At the same time, keratocytes are able to maintain nearly constant speed and direction during movement over many cell lengths. Their shapes, consisting of a bulbous cell body at the rear attached to a broad, thin lamellipodium at the front and sides, are simple, stereotyped and notoriously temporally persistent 9,10 . The molecular dynamism of these cells, combined with the persistence of their global shape and behaviour, make them an ideal model system for investigating the mechanisms of cell shape determination.The relative simplicity of keratocytes has inspired extensive experimental and theoretical investigations into this cell type 5-17 , considerably advancing the understanding of cell motility. A notable example is the graded radial extension (GRE) model 12 , which was an early attempt to link the mechanism of motility at the molecular level with overall cell geometry. The GRE model proposed that local cell extension (either protrusion or retraction) occurs perpendicular to the cell edge, and that the magnitude of this extension is graded from a maximum near the cell midline to a minimum towards the sides. Although this phenomenological model has been shown experimentally ...

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“…However, under certain cellular stress conditions, distinctive actin rods (also called bundles or paracrystals) can be induced in the nucleus in a variety of cell types. These conditions include dimethyl sulfoxide (DMSO) treatment (Sanger et al, 1980a;Sanger et al, 1980b), heat shock (Iida et al, 1986;Welch and Suhan, 1985), Latrunculin B treatment and ATP deletion (Pendleton et al, 2003) as well as viral infection (Charlton and Volkman, 1991;Feierbach et al, 2006). Cellular stress-induced formation of actin filaments seems to be caused by an increased nuclear actin level because nuclear translocation and accumulation of actin are also observed at the same time.…”

Section: Nuclear Architecture and Distribution Of Actinmentioning

confidence: 99%

“…Sanger and colleagues demonstrated that a disappearance of stress fibers from the cytoplasm and a reversible translocation of cytoplasmic actin into the nucleus occur after treatment of PtK2 and WI-38 cells with 10% DMSO (Sanger et al, 1980a;Sanger et al, 1980b). Courgeon and colleagues showed that heat shock causes actin to accumulate in the nucleus of Drosophila cells (Courgeon et al, 1993).…”

Section: Regulation Of Nuclear Translocation Of Actinmentioning

confidence: 99%

Exploring Secrets of Nuclear Actin Involvement in the Regulation of Gene Transcription and Genome Organization

Xu¹,

Kanagaratham²,

Radzioch³

2012

Current Frontiers and Perspectives in Cell Biology

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Dissolution of cytoplasmic actin bundles and the induction of nuclear actin bundles by dimethyl sulfoxide

Cited by 26 publications

References 15 publications

Reversible translocation of cytoplasmic actin into the nucleus caused by dimethyl sulfoxide.

Reversible translocation of cytoplasmic actin into the nucleus caused by dimethyl sulfoxide.

Mechanism of shape determination in motile cells

Exploring Secrets of Nuclear Actin Involvement in the Regulation of Gene Transcription and Genome Organization

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