Colchicine‐induced stimulation of PMN motility related to cytoskeletal changes in actin, α‐actinin, and myosin

Abstract

Colchicine-induced stimulation of polymorphonuclear leukocyte (PMN) locomotion is an interesting model because extension of blebs at the front occurs at a rate (about 2.4 μm/s) which is far above that reported for growth of actin filaments. The following cytoskeletal changes were observed in colchicine-treated PMNs: (1)a small increase in cytoskeleton-associated actin was noted, as well as a somewhate more pronounced increase in cytoskeleton-associated α-actinin, as compared with untreated or DMSO-treated controls. There was, however, no measurable increase in F-actin as determined by NBD-phallacidin blinding; (2)the values for the ratio (α-actinin/actin) are lower in PMNs treated with colchicine for 30 min, as compared with PMNs stimulated with fNLPNTL for 1 minute (non-polar ruffling cells) or 30 min (polarized locomoting cells); thus, this ratio may depend on the type of PMN motility; (3) in polarized PMNs F-actin was mainly located linearly all along the cell membrane; there was more intense staining at the front of the cells; (4) α-actining appeared to colocalize with F-actin at the leading front, but not with F-actin at the tail of polarized cells; (5) myosin was preferentially found at the rear part of polarized cells but not or only to a small extent at the front. Our data indicate a close functional correlation between microtubules and microfilaments. We speculate that F-actin in combination with α-actinin promotes expansion of pseudopods, whereas myosin combined with F-actin promotes contraction. In more general terms we suggest that different forms of PMN motility are generated by differential selective interaction of cytoskeletal compnents and variations in the composition of the cytoskeleton in different sites of the same cells.