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Biophys. J. BioFAST: First Published January 21, 2005. doi:10.1529/biophysj.104.046649
© 2005 by the Biophysical Society.

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CELL BIOPHYSICS

Probing the viscoelastic behavior of cultured airway smooth muscle cells with atomic force microscopy: stiffening induced by contractile agonist

Benjamin Smith 1*, Barbara Tolloczko 2, James G Martin 2 and Peter H Grütter 1

1 Department of Physics, McGill University
2 Department of Medicine, McGill University

* To whom correspondence should be addressed. E-mail: bsmith{at}physics.mcgill.ca.

Submitted on June 17, 2004
Revised on September 14, 2004
Accepted on 10 January 2005


  Abstract
Complex rheology of airway smooth muscle (ASM) cells, and its dynamic response during contractile stimulation, involves many molecular processes, foremost of which are actomyosin cross-bridge cycling and actin polymerization. With an atomic force microscope we tracked the spatial and temporal variations of the viscoelastic properties of cultured ASM cells. Elasticity mapping identified stiff structural elements of the cytoskeletal network. Using a precisely positioned microscale probe, picoNewton forces and nanometer level indentation modulations were applied to cell surfaces at frequencies ranging from 0.5 to 100 Hz. The resulting elastic storage modulus (G' ) and dissipative modulus (G'') increased dramatically, with hysteresivity ({eta} = G''/G') showing a definitive decrease following stimulation with the contractile agonist 5-hydroxytryptamine (5-HT). Frequency dependent assays showed weak power-law structural damping behavior and universal scaling in support of the soft-glassy material description of cellular biophysics. Additionally, a high frequency component of the loss modulus (attributed to cellular Newtonian viscosity)increased four-fold during the contractile process. The complex shear modulus showed a strong sensitivity to the degree of actin polymerization. Inhibitors of myosin light chain kinase (MLCK) activity had little effect on the stiffening response to contractile stimulation. Thus, our measurements appear to be particularly well suited for characterization of dynamic actin rheology during ASM contraction.

Key Words: actin polymerization, complex rheology, contractile stimulation, indentation modulation, myosin light chain kinase, soft glassy materials




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Copyright © 2005 by the Biophysical Society.