Astral microtubules physically redistribute cortical actin filaments to the incipient contractile ring

Abstract

Prior to cell cleavage, cytokinetic proteins are recruited into the nascent actomyosin contractile ring, paving the way for formation of a functional cleavage furrow. Interactions between spindle microtubules and the cell cortex may play a critical role in this recruitment, since microtubules have been shown to affect distribution and activation of cytokinetic proteins within the cortex. However, direct evidence for physical interaction between microtubules and the cortex has been lacking. Here, we probed the physical connection between astral microtubules and cortical actin filaments, by micromanipulating the fluorescently tagged cytoskeleton in living spermatocytes of the grasshopper Melanoplus femurrubrum. When microtubules were tugged with a microneedle, they in turn pulled on cortical actin filaments, interrupting the filaments' journey toward the equator. Further displacement of the actin dragged the cell membrane inward, demonstrating that the cortical actin network physically linked spindle microtubules to the cell membrane. Regional disruption of the connection by breaking spindle microtubules prevented actin accumulation in a segment of the ring, which locally inhibited furrowing. We propose a model in which dynamic astral microtubules physically redistribute cortical actin into the incipient contractile ring.