The final cytokinesis event involves severing of the connecting intercellular bridge (ICB) between daughter cells. FIP3-positive recycling endosomes (FIP3 endosomes) and ESCRT complexes have been implicated in mediating the final stages of cytokinesis. Here we analyse the spatiotemporal dynamics of the actin cytoskeleton, FIP3-endosome fusion and ESCRT-III localization during cytokinesis to show that the ICB narrows by a FIP3-endosome-mediated secondary ingression, whereas the ESCRT-III complex is needed only for the last scission step of cytokinesis. We characterize the role of FIP3 endosomes during cytokinesis to demonstrate that FIP3 endosomes deliver SCAMP2/3 and p50RhoGAP to the ICB during late telophase, proteins required for the formation of the secondary ingression. We also show that the FIP3-endosome-induced secondary ingression is required for the recruitment of the ESCRT-III complex to the abscission site. Finally, we characterize a FIP3-endosome-dependent regulation of the ICB cortical actin network through the delivery of p50RhoGAP. These results provide a framework for the coordinated efforts of actin, FIP3 endosomes and the ESCRTs to regulate cytokinesis and abscission.

Cytokinesis and abscission are complicated events that involve changes in membrane transport and cytoskeleton organization. We have used the combination of time-lapse microscopy and correlative high-resolution 3D tomography to analyze the regulation and spatio-temporal remodeling of endosomes and microtubules during abscission. We show that abscission is driven by the formation of a secondary ingression within the intracellular bridge connecting two daughter cells. The initiation and expansion of this secondary ingression requires recycling endosome fusion with the furrow plasma membrane and nested central spindle microtubule severing. These changes in endosome fusion and microtubule reorganization result in increased intracellular bridge plasma membrane dynamics and abscission. Finally, we show that central spindle microtubule reorganization is driven by localized microtubule buckling and breaking, rather than by spastin-dependent severing. Our results provide a new mechanism for mediation and regulation of the abscission step of cytokinesis.

Endocytic membrane transport has recently emerged as a key process required for the successful completion of cytokinesis. Specific endocytic membranes act in concert with the cytoskeleton and ESCRT proteins to regulate the various stages of cytokinesis. In this review, we focus on the different endocytic Arf and Rab GTPases and their interaction proteins that regulate organelle transport to the intracellular bridge during cytokinesis. The identity and function of these endocytic organelles during the late stages of cell division will also be discussed.