Regional rheological differences in locomoting neutrophils

1 Department of Geriatric and Respiratory Medicine, Tohoku University School of Medicine, Sendai 980-8574; 2 Center for Interdisciplinary Research, Tohoku Univ., Sendai 980; 3 Department of Metallurgy, Graduate School of Engineering, Tohoku University, Sendai 980, Japan; and 4 Harvard School of Public Health and Harvard Medical School, Boston, Massachusetts 02115 Submitted 12 August 2003 ; accepted in final form 26 March 2004 Intracellular rheology is a useful probe of the mechanisms underlying spontaneous or chemotactic locomotion and transcellular migration of leukocytes. We characterized regional rheological differences between the leading, body, and trailing regions of isolated, adherent, and spontaneously locomoting human neutrophils. We optically trapped intracellular granules and measured their displacement for 500 ms after a 100-nm step change in the trap position. Results were analyzed in terms of simple viscoelasticity and with the use of structural damping (stress relaxation follows a power law in time). Structural damping fit the data better than did viscoelasticity. Regional viscoelastic stiffness and viscosity or structural damping storage and loss moduli were all significantly lower in leading regions than in pooled body and/or trailing regions (the latter were not significantly different). Structural damping showed similar levels of elastic and dissipative stresses in body and/or trailing regions; leading regions were significantly more fluidlike (increased power law exponent). Cytoskeletal disruption with cytochalasin D or nocodazole made body and/or trailing regions 50% less elastic and less viscous. Cytochalasin D completely suppressed pseudopodial formation and locomotion; nocodazole had no effect on leading regions. Neither drug changed the dissipation-storage energy ratio. These results differ from those of studies of neutrophils and other cell types probed at the cell membrane via 2 -integrin receptors, which suggests a distinct role for the cell cortex or focal adhesion complexes. We conclude that 1 ) structural damping well describes intracellular rheology, and 2 ) while not conclusive, the significantly more fluidlike behavior of the leading edge supports the idea that intracellular pressure may be the origin of motive force in neutrophil locomotion. structural damping; power law; viscoelasticity; optical trap Address for reprint requests and other correspondence: M. Yanai, Dept. of Geriatric and Respiratory Medicine, Tohoku Univ.