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ABSTRACT: Transmigration through the endothelium is a key step in the immune response. In our recent work, the mechanical properties of the subendothelial matrix and biophysical state of the endothelium have been identified as key modulators of leukocyte trans-endothelial migration. Here, we demonstrated that neutrophil contractile forces and cytoskeletal dynamics also play an active biophysical role during transmigration through endothelial cell-cell junctions. Using our previously-established model for leukocyte transmigration, we first discovered that >93% of human neutrophils preferentially exploit the paracellular mode of transmigration in our in vitro model, and that is independent of subendothelial matrix stiffness. We demonstrated that inhibition of actin polymerization or depolymerization completely blocks transmigration, thus establishing a critical role for neutrophil actin dynamics in transmigration. Next, inhibition of neutrophil myosin II-mediated contractile forces renders 44% of neutrophils incapable of retracting their trailing edge under the endothelium for several minutes after the majority of the neutrophil transmigrates. Meanwhile, inhibition of neutrophil contractile forces or stabilization of microtubules doubles the time to complete transmigration for the first neutrophils to cross the endothelium. Notably, the time to complete transmigration is significantly reduced for subsequent neutrophils that cross through the same path as a previous neutrophil and is less dependent on neutrophil contractile forces and microtubule dynamics. These results suggest that the first neutrophil induces a gap in endothelial cell-cell adhesions, which "opens the door" in the endothelium and facilitates transmigration of subsequent neutrophils through the same hole. Collectively, this work demonstrates that neutrophils play an active biophysical role during the transmigration step of the immune response.
PLoS ONE 01/2013; 8(4):e61377. · 4.09 Impact Factor
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ABSTRACT: The immune response triggers a complicated sequence of events, one of which is release of the cytokine tumor necrosis factor-α (TNF-α) from stromal cells, for example monocytes and macrophages. In this work we investigated the biophysical effects of TNF-α on endothelial cells (ECs), including changes in cell morphology, biomechanics, migration, and cytoskeletal dynamics. We found that TNF-α induces a wide distribution of cell area and aspect ratio, with these properties increasing on average during treatment. Interestingly, aspect ratio peaks after approximately 10 h of exposure to TNF-α, corresponding also to a peak in exerted traction forces. Meanwhile, ECs treated with TNF-α soften, and we associate this with significant increases in estimated cellular volume. In addition, our evaluation of migratory dynamics revealed an inverse correlation between cell aspect ratio and migration speed after TNF-α treatment, suggesting that cell shape may be an important functional regulator of EC migration during an inflammatory response. Finally, we addressed the basic mechanics of how the reorganization of F-actin filaments occurs during TNF-α treatment, and observed a dynamic shift of existing actin filaments. Together, our results suggest a functional link between EC morphology, biomechanics, migration, and cytoskeletal dynamics during an inflammatory response.
Biophysics of Structure and Mechanism 09/2012; 41(11):939-47. · 2.44 Impact Factor
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ABSTRACT: Elevated levels of oxLDL in the bloodstream and increased vasculature stiffness are both associated with cardiovascular disease in patients. However, it is not known how oxLDL and subendothelial matrix stiffness together regulate an immune response. Here, we used an in vitro model of the vascular endothelium to explore the combined effects of oxLDL and subendothelial matrix stiffening on neutrophil transmigration. We prepared fibronectin-coated polyacrylamide gels of varying stiffness and plated human umbilical vein endothelial cells (ECs) onto the gels. We observed that oxLDL treatment of the endothelium promoted neutrophil transmigration (from <1% to 26% on soft 0.87kPa substrates), with stiffer substrates further promoting transmigration (54% on 5kPa and 41% on 280kPa). OxLDL exposure enhanced intercellular adhesion molecule-1 (ICAM-1) expression on the endothelium, which was likely responsible for the oxLDL-induced transmigration. Importantly, inhibition of MLCK-mediated EC contraction reduced transmigration to ∼9% on all substrates and eliminated the effects of subendothelial matrix stiffness. In addition, large holes, thousands of square microns in size, formed in monolayers on stiff substrates following transmigration, indicating that oxLDL treatment and subsequent neutrophil transmigration caused serious damage to the endothelium. Our results reveal that an interplay between ICAM-1 and MLCK-dependent contractile forces mediates neutrophil transmigration through oxLDL-treated endothelium. Thus, microvasculature stiffness, which likely varies depending on tissue location and health, is an important regulator of the transmigration step of the immune response in the presence of oxLDL.
Journal of biomechanics 05/2012; 45(10):1828-34. · 2.66 Impact Factor
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ABSTRACT: Changes in substrate compliance affect the cellular behavior of numerous cell types including epithelial, endothelial, fibroblasts,
and stem cells. Recently, an emphasis has been placed on understanding the mechanotactic behavior of neurons, in an attempt
to treat neurological injury and disease as well as to optimize the development of synthetic biomaterials for neural regeneration.
Here, we determine the stiffness of the fetal rat cortex using atomic force microscopy and evaluate the effect of substrate
mechanics on cortical neuron behavior using polyacrylamide gels with stiffness around that measured for the cortex. In particular,
we evaluate the relationship between substrate compliance and ligand coating to morphology, differentiation, and extension
behavior. Remarkably, we see an insensitivity of cortical process length and migration to substrate stiffness. We observe
differences in the tortuosity of process extension on laminin vs. poly-d-lysine, as well as differences in cell body migration; however these differences are independent of substrate compliance.
Myosin II inhibition revealed effects independent of stiffness, yet dependent on outgrowth behavior. Collectively, this work
suggests that cortical neurons are capable of differentiating and extending processes regardless of substrate stiffness, which
we attribute to the homogeneity of their native environment and their unwarranted need to distinguish substrate compliance.
KeywordsMechanotaxis-Axon differentiation-Polyacrylamide gels-Atomic force microscopy
Cellular and Molecular Bioengineering 04/2012; 3(4):398-414. · 1.95 Impact Factor
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ABSTRACT: Photopolymerizable phospholipid DC(8,9)PC (1,2-bis-(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine) exhibits unique assembly characteristics in the lipid bilayer. Because of the presence of the diacetylene groups, DC(8,9)PC undergoes polymerization upon UV (254 nm) exposure and assumes chromogenic properties. DC(8,9)PC photopolymerization in gel-phase matrix lipid 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) monitored by UV-vis absorption spectroscopy occurred within 2 min after UV treatment, whereas no spectral shifts were observed when DC(8,9)PC was incorporated into liquid-phase matrix 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). Liquid chromatography-tandem mass spectrometry analysis showed a decrease in the amount of DC(8,9)PC monomer in both DPPC and POPC environments without any change in the matrix lipids in UV-treated samples. Molecular dynamics (MD) simulations of DPPC/DC(8,9)PC and POPC/DC(8,9)PC bilayers indicate that the DC(8,9)PC molecules adjust to the thickness of the matrix lipid bilayer. Furthermore, the motions of DC(8,9)PC in the gel-phase bilayer are more restricted than in the fluid bilayer. The restricted motional flexibility of DC(8,9)PC (in the gel phase) enables the reactive diacetylenes in individual molecules to align and undergo polymerization, whereas the unrestricted motions in the fluid bilayer restrict polymerization because of the lack of appropriate alignment of the DC(8,9)PC fatty acyl chains. Fluorescence microscopy data indicates the homogeneous distribution of lipid probe 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-lissamine rhodamine B sulfonyl ammonium salt (N-Rh-PE) in POPC/DC(8,9)PC monolayers but domain formation in DPPC/DC(8,9)PC monolayers. These results show that the DC(8,9)PC molecules cluster and assume the preferred conformation in the gel-phase matrix for the UV-triggered polymerization reaction.
Langmuir 11/2011; 27(24):15120-8. · 4.19 Impact Factor
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ABSTRACT: A vast amount of work has been dedicated to the effects of shear flow and cytokines on leukocyte transmigration. However, no studies have explored the effects of substrate stiffness on transmigration. Here, we investigated important aspects of endothelial cell contraction-mediated neutrophil transmigration using an in vitro model of the vascular endothelium. We modeled blood vessels of varying mechanical properties using fibronectin-coated polyacrylamide gels of varying physiologic stiffness, plated with human umbilical vein endothelial cell (HUVEC) monolayers, which were activated with tumor necrosis factor-α. Interestingly, neutrophil transmigration increased with increasing substrate stiffness below the endothelium. HUVEC intercellular adhesion molecule-1 expression, stiffness, cytoskeletal arrangement, morphology, and cell-substrate adhesion could not account for the dependence of transmigration on HUVEC substrate stiffness. We also explored the role of cell contraction and observed that large holes formed in endothelium on stiff substrates several minutes after neutrophil transmigration reached a maximum. Further, suppression of contraction through inhibition of myosin light chain kinase normalized the effects of substrate stiffness by reducing transmigration and eliminating hole formation in HUVECs on stiff substrates. These results provide strong evidence that neutrophil transmigration is regulated by myosin light chain kinase-mediated endothelial cell contraction and that this event depends on subendothelial cell matrix stiffness.
Blood 06/2011; 118(6):1632-40. · 9.90 Impact Factor
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ABSTRACT: In this work we obtain the thermodynamic properties of mixed (1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine) PC and (1-stearoyl-2-oleoyl-sn-glycero-3-phospho-l-serine (sodium salt)) PS monolayers. Measurements of compressibility (isotherms, bulk modulus, and excess area per molecule) and surface potential show that the properties of monolayers at the air-water interface depend on the concentration of ions (Na(+) and K(+)) and the proportion of PS in the mixture. The dependence on PS arises because the molecule is originally bound to a Na(+) counterion; by increasing the concentration of ions the entropy changes, creating a favorable system for the bound counterions of PS to join the bulk, leaving a negatively charged molecule. This change leads to an increase in electrostatic repulsions which is reflected by the increase in area per molecule versus surface pressure and a higher surface potential. The results lead to the conclusion that this mixture of phospholipids follows a non ideal behavior and can help to understand the thermodynamic behavior of membranes made of binary mixtures of a zwitterionic and an anionic phospholipid with a bound counterion.
Colloids and surfaces. B, Biointerfaces 03/2011; 85(2):293-300. · 2.60 Impact Factor
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ABSTRACT: Biological processes such as atherogenesis, wound healing, cancer cell metastasis, and immune cell transmigration rely on a delicate balance between Cell-Cell and cell-substrate adhesion. Cell mechanics have been shown to depend on substrate factors such as stiffness and ligand presentation, while the effects of Cell-Cell interactions on the mechanical properties of cells has received little attention. Here, we use atomic force microscopy to measure the Young's modulus of live human umbilical vein endothelial cells (HUVECs). In varying the degree of Cell-Cell contact in HUVECs (single cells, groups, and monolayers), we observe that increased cell stiffness correlates with an increase in cell area. Further, we observe that HUVECs stiffen as they spread onto a glass substrate. When we weaken Cell-Cell junctions (i.e., through a low dose of cytochalasin B or treatment with a VE-cadherin antibody), we observe that cell-substrate adhesion increases, as measured by focal adhesion size and density, and the stiffness of cells within the monolayer approaches that of single cells. Our results suggest that while morphology can roughly be used to predict cell stiffness, Cell-Cell interactions may play a significant role in determining the mechanical properties of individual cells in tissues by careful maintenance of cell tension homeostasis.
Cellular and Molecular Bioengineering 03/2011; 4(1):9-27. · 1.95 Impact Factor
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ABSTRACT: Cell spreading is a critical component of numerous physiological phenomena including cancer metastasis, embryonic development, and mitosis. We have previously illustrated that cellular blebs appear after abrupt cell-substrate detachment and play a critical role in regulating membrane tension; however, the dynamics of bleb-substrate interactions during spreading remains unclear. Here we explore the role of blebs during endothelial cell spreading using chemical and osmotic modifications to either induce or inhibit bleb formation. We track cell-substrate dynamics as well as individual blebs using surface sensitive microscopic techniques. Blebbing cells (both control and chemically induced) exhibit increased lag times prior to fast growth. Interestingly, lamellae appear later for blebbing compared to non-blebbing cells, and in all cases, lamellae signal the start of fast spreading. Our results indicate that cellular blebs play a key role in the early stage of cell spreading, first by controling the initial cell adhesion and then by presenting a dynamic inhibition of cell spreading until a lamella appears and fast spreading ensues.
European journal of cell biology 11/2010; 90(1):37-48. · 3.31 Impact Factor
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ABSTRACT: Cells remodel their plasma membrane and cytoskeleton during numerous physiological processes, including spreading and motility. Morphological changes require the cell to adjust its membrane tension on different timescales. While it is known that endo- and exocytosis regulate the cell membrane area in a timescale of 1 h, faster processes, such as abrupt cell detachment, require faster regulation of the plasma membrane tension. In this article, we demonstrate that cell blebbing plays a critical role in the global mechanical homeostasis of the cell through regulation of membrane tension. Abrupt cell detachment leads to pronounced blebbing (which slow detachment does not), and blebbing decreases with time in a dynamin-dependent fashion. Cells only start spreading after a lag period whose duration depends on the cell's blebbing activity. Our model quantitatively reproduces the monotonic decay of the blebbing activity and accounts for the lag phase in the spreading of blebbing cells.
Biophysical Journal 09/2010; 99(6):1726-33. · 3.65 Impact Factor
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ABSTRACT: Room-temperature ionic liquids (ILs) exhibit a unique set of properties due to their charged character, presenting opportunities for numerous applications. Here, we show that the combination of charged surfactants with ILs leads to rich interfacial behavior due to the interplay between electrostatic and surface forces. Using traditional measures of surface activity and X-ray photoelectron spectroscopy (XPS), we find that sodium alkyl sulfates and alkyl trimethylammonium bromides are, indeed, surface-active at the air-IL interfaces of both [EMIM][EtSO(4)] and [BHEDMA][MeSO(3)]. XPS also reveals that surfactant counterions readily dissociate into the bulk, which when combined with the surfactant surface activity has striking consequences. We find that ion exchange occurs between surfactants and like-charged IL ions, with the greatest exchange for short surfactant alkyl chains. The initial negative surface charge of neat [EMIM][EtSO(4)] can be switched to positive by the addition of alkyl trimethylammonium bromides, with the effect most pronounced at short chain lengths. By contrast, the surface charge of [BHEDMA][MeSO(3)] is largely unaffected by the added surfactants, suggesting a key role for the strength of ion-pairing within the IL. The results here illustrate a simple but effective means of manipulating IL interfacial properties.
The Journal of Physical Chemistry B 09/2010; 114(35):11502-8. · 3.70 Impact Factor
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ABSTRACT: Cellular cholesterol is a critical component of the plasma membrane, and plays a key role in determining the physical properties of the lipid bilayer, such as elasticity, viscosity, and permeability. Surprisingly, it has been shown that cholesterol depletion increases cell stiffness, not due to plasma membrane stiffening, but rather, due to the interaction between the actin cytoskeleton and the plasma membrane. This indicates that traction stresses of the acto-myosin complex likely increase during cholesterol depletion. Here we use force traction microscopy to quantify the forces individual cells are exerting on the substrate, and total internal reflection fluorescence microscopy as well as interference reflection microscopy to observe cell-substrate adhesion and spreading. We show that single cells depleted of cholesterol produce larger traction forces and have large focal adhesions compared to untreated or cholesterol-enriched cells. Cholesterol depletion also causes a decrease in adhesion area for both single cells and monolayers. Spreading experiments illustrate a decrease in spreading area for cholesterol-depleted cells, and no effect on cholesterol-enriched cells. These results demonstrate that cholesterol plays an important role in controlling and regulating the cell-substrate interactions through the actin-plasma membrane complex, cell-cell adhesion, and spreading.
Cellular and Molecular Bioengineering 06/2010; 3(2):151-162. · 1.95 Impact Factor
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ABSTRACT: The vascular endothelium is exposed to an array of physical forces, including shear stress via blood flow, contact with other cells such as neighboring endothelial cells and leukocytes, and contact with the basement membrane. Endothelial cell morphology, protein expression, stiffness and cytoskeletal arrangement are all influenced by these mechanochemical forces. There are many biophysical tools that are useful in studying how forces are transmitted in endothelial cells, and these tools are also beginning to be used to investigate biophysical aspects of leukocyte transmigration, which is a ubiquitous mechanosensitive process. In particular, the stiffness of the substrate has been shown to have a significant impact on cellular behavior, and this is true for both endothelial cells and leukocytes. Thus, the stiffness of the basement membrane as an endothelial substrate, as well as the stiffness of the endothelium as a leukocyte substrate, is relevant to the process of transmigration. In this review, we discuss recent work that has related the biophysical aspects of endothelial cell interactions and leukocyte transmigration to the biochemical pathways and molecular interactions that take place during this process. Further use of biophysical tools to investigate the biological process of leukocyte transmigration will have implications for tissue engineering, as well as atherosclerosis, stroke and immune system disease research.
FEBS Journal 03/2010; 277(5):1145-58. · 3.79 Impact Factor
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ABSTRACT: Complete axon regeneration after trauma or disease is largely unsuccessful in the central nervous system. With the fast developing advances in tissue engineering and biomaterials, many investigations have identified promising approaches for guiding axonal extension. This review highlights a variety of these approaches and describes the biomaterial properties and signaling mechanisms involved in the fabrication of optimal guidance platforms. The vast majority of axonal regeneration approaches limit themselves to observe how axons elongate and migrate in response to signaling molecules presented on the substrate materials, or more recently, in response to different chemical and mechanical substrate properties. Many of these studies are encouraging in the hope of regenerating axons after disease or injury; however, numerous barriers remain. Here we illustrate the need to optimize a permissive heterogeneous environment for axon elongation using tissue engineering approaches and a thorough understanding of the mechanical properties of the substrate, mechanotaxis, and both attractive and repulsive signaling mechanisms.
Tissue Engineering Part B Reviews 06/2009; 15(3):291-305. · 4.64 Impact Factor
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ABSTRACT: Neutrophils are one type of migrating cell in the body's innate immune system and are the first line of defense against inflammation or infection. While extensive work exists on the effect of adhesive proteins on neutrophil motility, little is known about how neutrophil motility is affected by the mechanical properties of their physical environment. This study investigated the effects of substrate stiffness on the morphology, random motility coefficient, track speed (v), spreading area, and distribution of turning angles of neutrophils during chemokinesis. Human neutrophils were plated onto polyacrylamide gels of varying stiffness, ranging from 3 to 13 kPa, and coated with the extracellular matrix protein fibronectin, and timelapse images were taken with phase contrast microscopy. Our results show a biphasic behavior between neutrophil motility and substrate stiffness, with the optimum stiffness for motility depending on the concentration of fibronectin on the surface of the gel. On 100 microg/mL fibronectin, the optimum stiffness is 4 kPa (v = 6.9 +/- 0.6 microm/min) while on 10 microg/mL fibronectin, the optimum stiffness increases to 7 kPa (v = 4.5 +/- 2.0 microm/min). This biphasic behavior most likely arises because neutrophils on soft gels are less adherent, preventing production of traction forces, while neutrophils on stiff gels adhere strongly, resulting in decreased migration. At intermediate stiffness, however, neutrophils can attain optimal motility as a function of extracellular matrix coating.
Cell Motility and the Cytoskeleton 05/2009; 66(6):328-41. · 4.19 Impact Factor
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ABSTRACT: We find that in contrast to strongly adherent, slow moving cells such as fibroblasts, neutrophils exert contractile stresses largely in the rear of the cell (uropod) relative to the direction of motion. Rather than the leading edge pulling the cell, the rear is both anchoring the cell and the area in which the contractile forces are concentrated. These tractions rapidly reorient themselves during a turn, on a timescale of seconds to minutes, and their repositioning precedes and sets the direction of motion during a turn. We find the total average root mean-squared traction force to be 28+/-10 nN during chemokinesis, and 67+/-10 nN during chemotaxis. We hypothesize that the contraction forces in the back of the neutrophil not only break uropodial adhesive contacts but also create a rearward squeezing contractility, as seen in amoeboid or amoeboidlike cells and the formation of blebs in cells, causing a flow of intracellular material to the fluidlike lamellipod. Our findings suggest an entirely new model of neutrophil locomotion.
Biophysical Journal 05/2007; 92(7):L58-60. · 3.65 Impact Factor
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ABSTRACT: Leukocyte locomotion over the lumen of inflamed endothelial cells is a critical step, following firm adhesion, in the inflammatory response. Once firmly adherent, the cell will spread and will either undergo diapedesis through individual vascular endothelial cells or will migrate to tight junctions before extravasating to the site of injury or infection. Little is known about the mechanisms of neutrophil spreading or locomotion, or how motility is affected by the physical environment. We performed a systematic study to investigate the effect of the type of adhesive ligand and shear stress on neutrophil motility by employing a parallel-plate flow chamber with reconstituted protein surfaces of E-selectin, E-selectin/PECAM-1, and E-selectin/ICAM-1. We find that the level and type of adhesive ligand and the shear rate are intertwined in affecting several metrics of migration, such as the migration velocity, random motility, index of migration, and the percentage of cells moving in the direction of flow. On surfaces with high levels of PECAM-1, there is a near doubling in random motility at a shear rate of 180 s(-1) compared to the motility in the absence of flow. On surfaces with ICAM-1, neutrophil random motility exhibits a weaker response to shear rate, decreasing slightly when shear rate is increased from static conditions to 180 s(-1), and is only slightly higher at 1000 s(-1) than in the absence of flow. The random motility increases with increasing surface concentrations of E-selectin and PECAM-1 under static and flow conditions. Our findings illustrate that the endothelium may regulate neutrophil migration in postcapillary venules through the presentation of various adhesion ligands at sites of inflammation.
Biophysical Journal 02/2007; 92(2):632-40. · 3.65 Impact Factor
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ABSTRACT: Neutrophils rely on rapid changes in morphology to ward off invaders. Time-resolved dynamics of spreading human neutrophils after activation by the chemoattractant fMLF (formyl methionyl leucyl phenylalanine) was observed by RICM (reflection interference contrast microscopy). An image-processing algorithm was developed to identify the changes in the overall cell shape and the zones of close contact with the substrate. We show that in the case of neutrophils, cell spreading immediately after exposure of fMLF is anisotropic and directional. The dependence of spreading area, A, of the cell as a function of time, t, shows several distinct regimes, each of which can be fitted as power laws (A ~ t(b)). The different spreading regimes correspond to distinct values of the exponent b and are related to the adhesion state of the cell. Treatment with cytochalasin-B eliminated the anisotropy in the spreading.
Biophysical Journal 01/2007; 91(12):4638-48. · 3.65 Impact Factor
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ABSTRACT: This study has investigated the effect of cellular cholesterol on membrane deformability of bovine aortic endothelial cells. Cellular cholesterol content was depleted by exposing the cells to methyl-beta-cyclodextrin or enriched by exposing the cells to methyl-beta-cyclodextrin saturated with cholesterol. Control cells were treated with methyl-beta-cyclodextrin-cholesterol at a molar ratio that had no effect on the level of cellular cholesterol. Mechanical properties of the cells with different cholesterol contents were compared by measuring the degree of membrane deformation in response to a step in negative pressure applied to the membrane by a micropipette. The experiments were performed on substrate-attached cells that maintained normal morphology. The data were analyzed using a standard linear elastic half-space model to calculate Young elastic modulus. Our observations show that, in contrast to the known effect of cholesterol on membrane stiffness of lipid bilayers, cholesterol depletion of bovine aortic endothelial cells resulted in a significant decrease in membrane deformability and a corresponding increase in the value of the elastic coefficient of the membrane, indicating that cholesterol-depleted cells are stiffer than control cells. Repleting the cells with cholesterol reversed the effect. An increase in cellular cholesterol to a level higher than that of normal cells, however, had no effect on the elastic properties of bovine aortic endothelial cells. We also show that although cholesterol depletion had no apparent effect on the intensity of F-actin-specific fluorescence, disrupting F-actin with latrunculin A abrogated the stiffening effect. We suggest that cholesterol depletion increases the stiffness of the membrane by altering the properties of the submembrane F-actin and/or its attachment to the membrane.
Biophysical Journal 12/2004; 87(5):3336-43. · 3.65 Impact Factor
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ABSTRACT: Vesicles self-assembled from amphiphilic diblock copolymers exhibit a
wide diversity of behavior upon poration. This appears due to
competitive interplay between edge, surface and bending energies while
viscous dissipation mechanisms determine the time scales. The copolymers
studied are chemically identical, varying only in chain length which
dictates the membrane thickness d. For small d, we find large unstable
pores in electroporation, and the resulting membrane fragments
reassemble into vesicles within minutes. For large d, however, submicron
pores form and are extremely long-lived. The results show that pore
behavior depends strongly on d, suggesting that the relevant energies
depend on d and pore size r in a more complexmanner than what is
generally assumed. Qualitatively similar studies with lipid vesicles in
viscous glycerol have been performed by Brochard and coworkers, but the
present studies are simpler in not varying solvent.
02/2004; -1:31009.
