Signal Transduction in Smooth Muscle: Selected Contribution: Time course and heterogeneity of contractile responses in cultured human airway smooth muscle cells

1 Physiology Program, Harvard School of Public Health, Boston, Massachusetts 02115; 2 School of Biomedical Engineering, Dalhousie University, Halifax, Nova Scotia, Canada B3H 3J5; and 3 Pulmonary and Critical Care Division, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 We measured the time course and heterogeneity of responses to contractile and relaxing agonists in individual human airway smooth muscle (HASM) cells in culture. To this end, we developed a microrheometer based on magnetic twisting cytometry adapted with a novel optical detection system. Ferromagnetic beads (4.5 µm) coated with Arg-Gly-Asp peptide were bound to integrins on the cell surface. The beads were twisted in a sinusoidally varying magnetic field at 0.75 Hz. Oscillatory bead displacements were recorded using a phase-synchronized video camera. The storage modulus (cell stiffness; G'), loss modulus (friction; G"), and hysteresivity ( ; ratio of G" to G') could be determined with a time resolution of 1.3 s. Within 5 s after addition of histamine (100 µM), G' increased by 2.2-fold, G" increased by 3.0-fold, and increased transiently from 0.27 to 0.34. By 20 s, decreased to 0.25, whereas G' and G" remained above baseline. Comparable results were obtained with bradykinin (1 µM). These changes in G', G", and measured in cells were similar to but smaller than those reported for intact muscle strips. When we ablated baseline tone by adding the relaxing agonist dibutyryl cAMP (1 mM), G' decreased within 5 min by 3.3-fold. With relaxing and contracting agonists, G' could be manipulated through a contractile range of 7.3-fold. Cell populations exhibited a log-normal distribution of baseline stiffness (geometric SD = 2.8) and a heterogeneous response to both contractile and relaxing agonists, partly attributable to variability of baseline tone between cells. The total contractile range of the cells (from maximally relaxed to maximally stimulated), however, was independent of baseline stiffness. We conclude that HASM cells in culture exhibit a clear, although heterogeneous, response to contractile and relaxing agonists and express the essential mechanical features characteristic of the contractile response observed at the tissue level. cell mechanics; muscle contraction; muscle relaxation; actin; myosin; bridge dynamics; cytoskeleton