Co-movement of astral microtubules, organelles and F-actin suggests aster positioning by surface forces in frog eggs

Abstract

AbstractHow bulk cytoplasm generates forces to separate post-anaphase microtubule (MT) asters in Xenopus laevis and other large eggs remains unclear. Previous models proposed dynein-based organelle transport generates length-dependent forces on astral MTs that pull centrosomes through the cytoplasm, away from the midplane. In Xenopus egg extracts, we co-imaged MTs, endoplasmic reticulum (ER), mitochondria, acidic organelles, F-actin, keratin, and fluorescein in moving and stationary asters. In asters that were moving in response to dynein and actomyosin forces, we observed that all cytoplasmic components moved together, i.e., as a continuum. Dynein-mediated organelle transport was restricted by interior MTs and F-actin. Organelles exhibited a burst of dynein-dependent inward movement at the growing aster surface, then mostly halted inside the aster. Dynein-coated beads were slowed by F-actin, but in contrast to organelles, beads did not halt inside asters. These observations call for new models of aster positioning based on surface forces and internal stresses.