Theoretical Prediction of the Orientation of Cultured Cells and Intracellular Stress Fibers under Biaxial Cyclic Deformations

2002 ◽  
Vol 2002.14 (0) ◽  
pp. 259-260
Author(s):  
Hiroshi YAMADA ◽  
Daisaku MORITA ◽  
Tohru TAKEMASA ◽  
Takami YAMAGUCHI
Keyword(s):  
Theoretical Prediction ◽  
Cultured Cells ◽  
Stress Fibers

scholarly journals Microinjection of nonmuscle and smooth muscle caldesmon into fibroblasts and muscle cells.

1990 ◽  
Vol 111 (6) ◽  
pp. 2487-2498 ◽  
Author(s):  
Y Yamakita ◽  
S Yamashiro ◽  
F Matsumura
Keyword(s):  
Smooth Muscle ◽  
Molecular Mass ◽  
Cultured Cells ◽  
Muscle Cells ◽  
Biochemical Properties ◽  
Stress Fibers ◽  
Relative Molecular Mass ◽  
Short Period ◽  
Nonmuscle Cells ◽  
Mass Form

Caldesmon is present in a high molecular mass form in smooth muscle and predominantly in a low molecular mass form in nonmuscle cells. Their biochemical properties are very similar. To examine whether these two forms of caldesmon behave differently in cultured cells, we microinjected fluorescently labeled smooth muscle and nonmuscle caldesmons into fibroblasts. Simultaneous injection of both caldesmons into the same cells has revealed that both high and low relative molecular mass caldesmons are quickly (within 10 min) and stably (over 3 d) incorporated into the same structures of microfilaments including stress fibers and membrane ruffles, suggesting that nonmuscle cells do not distinguish nonmuscle caldesmon from smooth muscle caldesmon. The effect of calmodulin on the incorporation of caldesmon has been examined by coinjection of caldesmon with calmodulin. We have found that calmodulin retards the incorporation of caldesmon into stress fibers for a short period (10 min) but not for a longer incubation (30 min). The behavior of caldesmon in developing muscle cells was also examined because we previously observed that caldesmon disappears during myogenesis (Yamashiro, S., R. Ishikawa, and F. Matsumura. 1988. Protoplasma Suppl. 2: 9-21). We have found that, in contrast to its stable incorporation into stress fibers of fibroblasts, caldesmon is unable to be incorporated into thin filament structure (I-band) of differentiated muscle.

Macromolecular specializations that mediate lateral interactions between microfilaments and the membrane at focal contacts

1992 ◽  
Vol 50 (1) ◽  
pp. 724-725
Author(s):  
Steven J. Samuelsson ◽  
Paul W. Luther ◽  
David W. Pumplin ◽  
Robert J. Bloch
Keyword(s):  
Immunofluorescence Microscopy ◽  
Cultured Cells ◽  
Stress Fiber ◽  
Stress Fibers ◽  
Focal Contact ◽  
Lateral Interactions ◽  
Matrix Assembly ◽  
Cytoplasmic Surface ◽  
Focal Contacts ◽  
Specific Proteins

Focal contacts are membrane specializations of cultured cells where stress fibers terminate and where the cell is most closely applied to the substrate. The organization of this cytoskeletal-membrane-extracellular matrix assembly has been well characterized. Immunofluorescence microscopy has shown that two focal contact-specific proteins, vinculin and talin, colocalize with microfilaments for several microns before the stress fiber terminates. This result raises the question of whether microfilament-membrane interactions are limited to the ends of microfilaments, or if lateral interactions also occur. We addressed this question by examining the cytoplasmic surface of isolated focal contacts in detail.

Relationship Between Orientation Angle in Intracellular Stress Fibers and Range of Uniaxial Cyclic Stretch

2000 ◽  
Author(s):  
Hiroshi Yamada ◽  
Tohru Takemasa ◽  
Takami Yamaguchi
Keyword(s):  
Endothelial Cells ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Orientation Angle ◽  
Mechanical Stimulus ◽  
Cyclic Stretch ◽  
Stress Fibers ◽  
Experimental Result ◽  
Silicone Membrane ◽  
Umbilical Vein Endothelial Cells

Abstract We hypothesized an avoidance of deformation and a limit of sensitivity of cell response to the mechanical stimulus for the orientation of stress fibers in cultured cells on a silicone membrane which was subjected to a uniaxial cyclic stretch. We compared the theoretical prediction with the experimental result of stress fibers in the human umbilical vein endothelial cells under various ranges of uniaxial cyclic stretch (Takemasa et al. 1998). The results showed that the proposed hypothesis predicted the orientation of stress fibers under various ranges of cyclic stretch well.

scholarly journals Mobility of filamentous actin in living cytoplasm.

1987 ◽  
Vol 105 (6) ◽  
pp. 2811-2816 ◽  
Author(s):  
Y L Wang
Keyword(s):  
Cell Division ◽  
Cultured Cells ◽  
Stress Fibers ◽  
Detectable Effect ◽  
Filamentous Actin ◽  
Cellular Functions ◽  
Contractile Ring ◽  
Rhodamine Phalloidin ◽  
Affinity Probe ◽  
Dividing Cells

Filamentous actin in living cultured cells was labeled by microinjecting trace amounts of rhodamine-phalloidin (rh-pha) as a specific, high-affinity probe. The microinjection caused no detectable effect on cell morphology or cell division. The distribution of rh-pha-labeled filaments was then examined in dividing cells using image-intensified fluorescence microscopy, and the exchangeability of labeled filaments along stress fibers was studied during interphase using fluorescence recovery after photobleaching. rh-pha showed a rapid concentration at the contractile ring during cell division. In addition, recovery of fluorescence after photobleaching occurred along stress fibers with a halftime as short as 8 min. These observations suggest that at least some actin filaments undergo continuous movement and reorganization in living cells. This dynamic process may play an important role in various cellular functions.

scholarly journals Fibronectin Regulates Assembly of Actin Filaments and Focal Contacts in Cultured Cells via the Heparin-binding Site in Repeat III13

1999 ◽  
Vol 10 (5) ◽  
pp. 1521-1536 ◽  
Author(s):  
Laird Bloom ◽  
Kenneth C. Ingham ◽  
Richard O. Hynes
Keyword(s):  
Matrix Protein ◽  
Cultured Cells ◽  
Fiber Formation ◽  
Stress Fibers ◽  
Extracellular Matrix Protein ◽  
Heparin Binding ◽  
Actin Organization ◽  
Focal Contacts ◽  
Α5β1 Integrin ◽  
Amino Acid Segment

Fibroblasts, when plated on the extracellular matrix protein fibronectin (FN), rapidly spread and form an organized actin cytoskeleton. This process is known to involve both the central α5β1 integrin-binding and the C-terminal heparin-binding regions of FN. We found that within the heparin-binding region, the information necessary for inducing organization of stress fibers and focal contacts was located in a 29–amino acid segment of FN type III module 13 (III13). We did not find a cytoskeleton-organizing role for repeat III14, which had previously been implicated in this process. Within III13, the same five basic amino acids known to be most important for heparin binding were also necessary for actin organization. A substrate of III13 alone was only weakly adhesive but strongly induced formation of filopodia and lamellipodia. Stress fiber formation required a combination of III13 and III7–11(which contains the integrin α5β1 recognition site), either as a single fusion protein or as separate polypeptides, and the relative amounts of the two binding sites appeared to determine whether stress fibers or filopodia and lamellipodia were the predominant actin structures formed. We propose that a balance of signals from III13 and from integrins regulates the type of actin structures assembled by the cell.

Vasoactive agents affect mesangial cell adhesion

1986 ◽  
Vol 251 (4) ◽  
pp. C505-C511 ◽  
Author(s):  
J. I. Kreisberg ◽  
M. A. Venkatachalam
Keyword(s):  
Cell Adhesion ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Mesangial Cell ◽  
Cultured Cells ◽  
Mesangial Cells ◽  
Stress Fibers ◽  
Vasoactive Agents ◽  
Substrate Adhesion ◽  
Adhesive Contacts

The formation and maintenance of stress fibers in cultured mesangial cells is associated with myosin light chain phosphorylation [Kreisberg et al. Am. J. Physiol. 249 (Renal Fluid Electrolyte Physiol. 18): F227-F235, 1985], a biochemical indicator for activation of actin-myosin interactions. Agents that elevate intracellular levels of adenosine 3',5'-cyclic monophosphate (cAMP) (e.g., isoproterenol) fragment stress fibers and cause myosin light chain dephosphorylation, whereas the addition of contractile agents such as arginine vasopressin (AVP) and prostaglandin E2 (PGE2) reverses these changes. Because stress fiber development in cultured cells is correlated with tight cell to substrate adhesion, we wanted to examine whether vasoactive agents have an effect on mesangial cell adhesion. Both isoproterenol and dibutyryl cAMP (DBcAMP) reduced mesangial cell adherence as measured by a trypsin assay (% detached cells: control 11 +/- 2.4%; isoproterenol plus isobutylmethylxanthine (IBMX) = 48.3 +/- 7.4%; DBcAMP = 29.3 +/- 3.7%; DBcAMP-IBMX = 73 +/- 4.4%). The areas of focal (adhesive) contacts between the cell and substratum as observed by interference-reflexion microscopy were also reduced, being replaced by areas of greater separation (% of the surface in contact with the substratum: control = 7.4 +/- 0.8%; isoproterenol-IBMX = 2.9 +/- 1.1%). Addition of PGE2 or AVP to the incubation medium containing the cAMP-elevating agents prevented the above changes. PGE2 or AVP alone increased mesangial cell adhesion (% detached cells: control 11 +/- 2.4%; PGE2 = 6.8 +/- 0.5%; AVP = 5.1 +/- 1.2%).(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Characterization of Palladin, a Novel Protein Localized to Stress Fibers and Cell Adhesions

2000 ◽  
Vol 150 (3) ◽  
pp. 643-656 ◽  
Author(s):  
Mana M. Parast ◽  
Carol A. Otey
Keyword(s):  
Cultured Cells ◽  
Focal Adhesions ◽  
Cell Types ◽  
Cell Junctions ◽  
Stress Fibers ◽  
Western Blots ◽  
Embryonic Tissues ◽  
Mouse Tissues ◽  
Antisense Constructs

Here, we describe the identification of a novel phosphoprotein named palladin, which colocalizes with α-actinin in the stress fibers, focal adhesions, cell–cell junctions, and embryonic Z-lines. Palladin is expressed as a 90–92-kD doublet in fibroblasts and coimmunoprecipitates in a complex with α-actinin in fibroblast lysates. A cDNA encoding palladin was isolated by screening a mouse embryo library with mAbs. Palladin has a proline-rich region in the NH2-terminal half of the molecule and three tandem Ig C2 domains in the COOH-terminal half. In Northern and Western blots of chick and mouse tissues, multiple isoforms of palladin were detected. Palladin expression is ubiquitous in embryonic tissues, and is downregulated in certain adult tissues in the mouse. To probe the function of palladin in cultured cells, the Rcho-1 trophoblast model was used. Palladin expression was observed to increase in Rcho-1 cells when they began to assemble stress fibers. Antisense constructs were used to attenuate expression of palladin in Rcho-1 cells and fibroblasts, and disruption of the cytoskeleton was observed in both cell types. At longer times after antisense treatment, fibroblasts became fully rounded. These results suggest that palladin is required for the normal organization of the actin cytoskeleton and focal adhesions.

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