Microtubules can modulate pseudopod activity from a distance inside macrophages.

Abstract

Microtubules are thought to influence cell shape as structural components of an integrated cytoskeletal matrix. Here we show that microtubules can affect the dynamics of macrophage pseudopodia without being integrated into their structure. Macrophages landing on glass surfaces spread within 15 min into flattened circular cells with radial symmetry, and the radial distribution of microtubules reflected this symmetry. Depolymerization of microtubules using nocodazole, colchicine, or vinblastine did not inhibit macrophage spreading or the early establishment of radial symmetry. Shortly after spreading, however, macrophages without microtubules gradually became asymmetric, assuming irregular, lobed profiles. The asymmetry resulted from exaggerated protrusion and retraction of pseudopodia, with net retraction overall. This loss of radial symmetry could be inhibited by treatment of initially spread cells with cytochalasin D, indicating that the change in cell shape was mediated by the actin cytoskeleton. Intact microtubules suppressed the exaggerated pseudopod movements, even when they were separated by a distance from the cell margin. In cells treated with taxol, microtubules remained clustered near the cell center after spreading, yet the dynamics of pseudopodia at the cell margin were reduced and cells maintained a circular profile. Similarly, in cells treated with low concentrations of nocodazole, a much reduced microtubule cytoskeleton nonetheless suppressed pseudopod dynamics. We propose that microtubules act to stabilize cell shape at a distance from the cell edge via a biochemical intermediate that affects the structure or function of the microfilament system.