Cytoplasmic actomyosin fibrils in tissue culture cells

Isenberg, G.; Rathke, P. C.; Hülsmann, Norbert; Franke, Werner W.; Wohlfarth‐Bottermann, K. E.

doi:10.1007/bf00225909

Cited by 157 publications

(21 citation statements)

References 33 publications

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“…There is evidence that in vitro stress fibers are related to isometric contraction and adhesion to the substrate rather than to isotonic contraction and cell movement (26), despite the fact that isolated stress fibers can contract under appropriate conditions (27,28). In our experiments, stress fibers appear in moving and replicating cells but persist long after endothelial cell progression toward the denuded area and replication have ceased (18).…”

Section: Discussionmentioning

confidence: 52%

Organization of actin cytoskeleton in normal and regenerating arterial endothelial cells.

Gabbiani¹,

Gabbiani²,

Lombardi³

1983

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

106

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The distribution of actin stress fibers in normal and regenerating (after endothelial denudation by means of a balloon catheter) rabbit aortic endothelial cells has been studied by means of immunofluorescence with human actin autoantibodies on enface endothelial cell preparations. Our results show that: (i) under normal conditions actin is accumulated as a network at the periphery of endothelial cells. Stress fibers are present only in endothelial cells located immediately below intercostal artery branches; (ii) stress fibers develop in endothelial cells early during regeneration and persist after the end of endothelial mitotic and motile activities; and (iii) the orientation of stress fibers within the cytoplasm follows the direction of blood flow, with the exception of stress fibers situated in cells at the edge of the wound, when endothelial cell progression toward the denuded area as well as mitotic activity have ceased. We conclude that stress fibers are an organelle present in endothelial cells in vivo and that they reorganize during endothelial cell adaptation to unfavorable or pathological situations.Endothelial continuity depends on the maintenance of an extremely thin single cell layer. When arterial endothelium is removed by mechanical abrasion, the wound is repaired by endothelial cell replication and movement (1). When the endothelial cell loss is small (few cells in width) repair is accomplished essentially by movement without replication (2). Cell movement results probably from the activity of cytoskeletal and cytocontractile elements (3), among which actin is the most abundant. Cellular actin (4, 5) is organized in the form of G actin, a microfilamentous network, and microfilamentous bundles or stress fibers, which are typical of most adherent cultured cells, including endothelial cells. Stress fibers have been shown to contain actin, myosin, and regulatory proteins (6-9). Little information is presently available on the distribution of stress fibers in vivo (5).We have studied the organization of stress fibers in endothelial cells of rabbit aorta and carotid artery under normal conditions and during regeneration after ballooning-induced endothelial denudation (10). For this purpose, we have developed a technique of immunofluorescent staining with antiactin antibodies (AAAbs) on en face endothelial cell preparations. Our results show that under normal conditions stress fibers are present only in some aortic endothelial cells-namely, those submitted to high shear stress; they become common in regenerating endothelial cells and persist long after motile and mitotic activities have ceased. MATERIALS AND METHODSMale rabbits, -3 kg of body weight, were used for the experiments. The endothelium was removed from the thoracic aorta or the left carotid artery by using the technique originally described by Baumgartner and Studer (10) with minor modifications. Evans blue (4 ml of a 1% solution in 0.9% NaCI) was injected intravenously 20 min prior to sacrifice to label denuded areas (blue areas). Tw...

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Section: Discussionmentioning

confidence: 52%

Organization of actin cytoskeleton in normal and regenerating arterial endothelial cells.

Gabbiani¹,

Gabbiani²,

Lombardi³

1983

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

106

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“…Given these observations and the evidence that many stress fibers terminate on substrate attachment plaques (17,27), it seems likely that most stress fibers have a structural role in anchoring the cytoplasmic matrix to the substrate, rather than being contractile . Their contractile potential is expressed only when one or both of their terminal attachment plaques is released from the substrate, such as in experimental manipulation (26,28) . Actually, neither the function of these photogenic animyosin bundles in vitro nor their very existence in vivo is established .…”

Section: Discussionmentioning

confidence: 99%

Relation between cell activity and the distribution of cytoplasmic actin and myosin.

Herman¹,

Crisona²,

Pollard³

1981

The Journal of Cell Biology

219

103

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We documented the activity of cultured cells on time-lapse videotapes and then stained these identified cells with antibodies to actin and myosin. This experimental approach enabled us to directly correlate cellular activity with the distribution of cytoplasmic actin and myosin. When trypsinized HeLa cells spread onto a glass surface, the cortical cytoplasm was the most actively motile and random, bleb-like extensions (0.5-4.0 micrometer wide, 2-5 micrometer long) occurred over the entire surface until the cells started to spread. During spreading, ruffling membranes were found at the cell perimeter. The actin staining was found alone in the surface blebs and ruffles and together with myosin staining in the cortical cytoplasm at the bases of the blebs and ruffles. In well-spread, stationary HeLa cells most of the actin and myosin was found in stress fibers but there was also diffuse antiactin fluorescence in areas of motile cytoplasm such as leading lamellae and ruffling membranes. Similarly, all 22 of the rapidly translocating embryonic chick cells had only diffuse actin staining. Between these extremes were slow-moving HeLa cells, which had combinations of diffuse and fibrous antiactin and antimyosin staining. These results suggest that large actomyosin filament bundles are associated with nonmotile cytoplasm and that actively motile cytoplasm has a more diffuse distribution of these proteins.

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“…As a result ofmany studies, it is fairly well accepted that the cytoskeleton is a major element in the cellular motile apparatus (1-5). Cytoskeletal proteins such as actin and myosin are thought to generate forces necessary-for movement (5)(6)(7)(8). In addition to their role in cell division, microtubules (tubulin and associated proteins) have been studied as support structures involved in maintaining cell shape and polarity (5, 9, 10), intracellular partical transport (11), and as structures involved in directed movement of leukocytes (1,12,13) .…”

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confidence: 99%

Laser irradiation of centrosomes in newt eosinophils: evidence of centriole role in motility.

Koonce¹,

Cloney²,

Berns³

1984

The Journal of Cell Biology

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Newt eosinophils are motile granulated leukocytes that uniquely display a highly visible centrosomal area. Electron microscope and tubulin antibody fluorescence confirms the presence of centrioles, pericentriolar material, and radiating microtubules within this visible area . Actin antibodies intensely stain the advancing cell edges and tail but only weakly stain pseudopods being withdrawn into the cell. Randomly activated eosinophils follow a roughly consistant direction with an average rate of 22 .5 gm/min . The position of the centrosome is always located between the trailing cell nucleus and advancing cell edge . If the cell extends more than one pseudopod, the one closest to or containing the centrosome is always the one in which motility continues .Laser irradiation of the visible centrosomal area resulted in rapid cell rounding . After several minutes following irradiation, most cells flattened and movement continued. However, postirradiation motility was uncoordinated and directionless, and the rate decreased to an average of 14 .5 Am/min . Electron microscopy and tubulin immunofluorescence indicated that an initial disorganization of microtubules resulted from the laser microirradiations . After several minutes, organized microtubules reappeared, but the centrioles appeared increasingly damaged . The irregularities in motility due to irradiation are probably related to the damaged centrioles. The results presented in this paper suggest that the centrosome is an important structure in controlling the rate and direction of newt eosinophil motility .Cell movement is a fundamental process of particular relevance to problems in developmental biology, cell biology, oncology, and immunology. Recent advances in immunofluorescence and electron microscopy have resulted in considerable research on the mechanisms ofcell motility. As a result ofmany studies, it is fairly well accepted that the cytoskeleton is a major element in the cellular motile apparatus (1-5). Cytoskeletal proteins such as actin and myosin are thought to generate forces necessary-for movement (5)(6)(7)(8). In addition to their role in cell division, microtubules (tubulin and associated proteins) have been studied as support structures involved in maintaining cell shape and polarity (5, 9, 10), intracellular partical transport (11), and as structures involved in directed movement of leukocytes (1,12,13) . Recently, the microtubule-organizing centers have been suggested as the structures involved in direction determination (14-17) .

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Cytoplasmic actomyosin fibrils in tissue culture cells

Cited by 157 publications

References 33 publications

Organization of actin cytoskeleton in normal and regenerating arterial endothelial cells.

Organization of actin cytoskeleton in normal and regenerating arterial endothelial cells.

Relation between cell activity and the distribution of cytoplasmic actin and myosin.

Laser irradiation of centrosomes in newt eosinophils: evidence of centriole role in motility.

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