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Biophys J, June 2001, p. 2649-2657, Vol. 80, No. 6
and
*Physik Department E22 (Biophysics group), Technische
Universität München, D-85748 Garching; Max
von Pettenkofer Institut für Medizinische Mikrobiologie,
Universität München, Pettenkoferstr. 9a, 80336 München, Instutut für Prophylaxe und
Epidemiologie der Kreislaufkrankheiten, Universität
München, Pettenkoferstr. 9, D-80336 München, Germany
By using magnetic bead microrheology we study the effect
of inflammatory agents and toxins on the viscoelastic moduli of
endothelial cell plasma membranes in real time. Viscoelastic response
curves were acquired by applying short force pulses of ~500 pN to
fibronectin-coated magnetic beads attached to the surface membrane of
endothelial cells. Upon addition of thrombin, a rapid stiffening of the
membrane was observed within 5 s, followed by recovery of the
initial deformability within 2 min. By using specific inhibitors, two
known pathways by which thrombin induces actin reorganization in
endothelial cells, namely activation of
Ca2+-calmodulin-dependent myosin light chain kinase and
stimulation of Rho/Rho-kinase, were excluded as possible causes of the
stiffening effect. Interestingly, the cytotoxic necrotizing factor of
Escherichia coli, a toxin which, in addition to Rho,
activates the GTPases Rac and CDC42Hs, also induced a dramatic
stiffening effect, suggesting that the stiffening may be mediated
through a Rac- or Cdc42Hs-dependent pathway. This work demonstrates
that magnetic bead microrheometry is not only a powerful tool to
determine the absolute viscoelastic moduli of the composite cell plasma
membrane, but also a valuable tool to study in real time the effect of
drugs or toxins on the viscoelastic parameters of the plasma membrane.
Biophys J, June 2001, p. 2649-2657, Vol. 80, No. 6
© 2001 by the Biophysical Society 0006-3495/01/06/2649/09 $2.00
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