ABSTRACT
AIM: To investigate the link between cell stiffness and volume-regulated anion current (VRAC) in aortic endothelium. METHOD: Bovine aortic endothelial cells (BAECs) were exposed to methyl-beta-cyclodextrin (MbetaCD) to deplete cellular cholesterol and the changes in cellular stiffness were measured by micropipette aspiration. VRAC density was measured electrophysiologically in the same cell populations. Furthermore, to probe the effects of cholesterol depletion on the mechanics of 'deep' cytoskeleton, we employ a novel technique to analyse correlated motion of intracellular particles. RESULTS: We show that cholesterol depletion results in cellular stiffening and an upregulation of VRAC density. Replenishing cellular sterol pool with epicholesterol, a chiral analogue of cholesterol, abrogates both of these effects. This indicates that cholesterol sensitivity of both cell mechanics and VRAC are due to changes in the physical properties of the membrane rather than due to specific sterol-protein interactions. We also show that cholesterol depletion increases the stiffness of the 'deep cytoskeleton' and that disruption of actin filaments abolishes both cell stiffening and upregulation of VRAC due to cholesterol depletion. Furthermore, comparing BAECs to human aortic endothelial cells (HAECs), we show that BAECs that are inherently stiffer also develop larger VRACs. CONCLUSIONS: Taken together, our observations suggest an increase in the cytoskeleton stiffness has a facilitatory effect on VRAC development. We suggest that stiffening of the cytoskeleton increases tension in the membrane-cytoskeleton layer and that in turn facilitates VRAC.
Subject(s)
Cytoskeleton/physiology , Endothelial Cells/physiology , Endothelium, Vascular/physiology , Ion Channels/metabolism , Animals , Anticholesteremic Agents , Aorta , Cattle , Cell Membrane/physiology , Cell Size , Cells, Cultured , Cholesterol/analysis , Cholesterol/metabolism , Cytoskeleton/metabolism , Elasticity , Electrophysiology , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Humans , Lipid Bilayers , Micromanipulation , beta-Cyclodextrins/pharmacologyABSTRACT
We exploit the power of microrheology to measure the viscoelasticity of entangled F-actin solutions at different length scales from 1 to 100 microm over a wide frequency range. We compare the behavior of single probe-particle motion to that of the correlated motion of two particles. By varying the average length of the filaments, we identify fluctuations that dissipate diffusively over the filament length. These provide an important relaxation mechanism of the elasticity between 0.1 and 30 rad/sec.
Subject(s)
Actins/chemistry , Elasticity , Polyethylene Glycols , Polystyrenes , Rheology , Solutions , ViscosityABSTRACT
We report the first measurements of the intrinsic strain fluctuations of living cells using a recently developed tracer correlation technique along with a theoretical framework for interpreting such data in heterogeneous media with nonthermal driving. The fluctuations' spatial and temporal correlations indicate that the cytoskeleton can be treated as a course-grained continuum with power-law rheology, driven by a spatially random stress tensor field. Combined with recent cell rheology results, our data imply that intracellular stress fluctuations have a nearly 1/omega2 power spectrum, as expected for a continuum with a slowly evolving internal prestress.
Subject(s)
Cell Physiological Phenomena , Rheology/methods , Elasticity , Mathematical Computing , Stress, Mechanical , ViscosityABSTRACT
This report describes repair of an aortoesophageal fistula caused by a previously placed thoracic aortic graft. The diagnosis was made by esophagoscopy. The repair consisted of femoral-to-femoral cardiopulmonary bypass, excision of the old graft, placement of a new graft, esophagectomy, cervical esophagostomy, gastrostomy, and later reconstruction by cervical esophagogastrostomy.
Subject(s)
Aorta/surgery , Aortic Diseases/surgery , Blood Vessel Prosthesis , Esophageal Fistula/surgery , Fistula/surgery , Postoperative Complications , Esophagectomy , Esophagostomy , Female , Gastrostomy , Humans , Middle AgedABSTRACT
The effects of three compounds on the cell cycle of HL-60 promyeloid leukemia cells has been examined. Ciclopirox olamine, an antifungal agent, and the compound Hoechst 768159 reversibly block the cell cycle at a point occurring roughly 1 h before the arrest mediated by aphidicolin, an inhibitor of DNA polymerase alpha activity, which acts in early S phase. Similar results are also obtained with the compound mimosine, a plant amino acid. Based on these data, it is concluded that all three agents inhibit cell cycle traverse at or very near the G1/S phase boundary and identify a previously undefined reversible cell cycle arrest point.
