Aparna Sherlekar

Branched actin networks driven by Arp2/3 collaborate with actomyosin filaments in processes such as cell migration. The syncytial Drosophila blastoderm embryo also shows expansion of apical caps by Arp2/3 driven actin polymerization in interphase and buckling at contact edges by MyosinII to form furrows in metaphase. Here we study the role of Syndapin (Synd), an F-BAR domain containing protein in apical cap remodelling prior to furrow extension. synd depletion showed larger apical caps. STED super-resolution and TIRF microscopy showed long apical actin protrusions in caps in interphase and short protrusions in metaphase in control embryos. synd depletion led to sustained long protrusions even in metaphase. Loss of Arp2/3 function in synd mutants partly reverted defects in apical cap expansion and protrusion remodelling. MyosinII levels were decreased in synd mutants and MyosinII mutant embryos have been previously reported to have expanded caps. We propose that Syndapin function limits branching activity during cap expansion and affects MyosinII distribution in order to shift actin remodeling from apical cap expansion to favor lateral furrow extension.

BAR domain proteins can regulate ‘membrane reservoirs’ that provide surface area and buffer membrane tension. Syndapin is an F-BAR and SH3 domain containing protein involved in cytoskeletal remodelling and endocytosis. The Syndapin F-BAR domain is uniquely versatile compared to others in the family and can bend phospholipid membranes into tubules of various diameters and directly bind actin. The Syndapin SH3 domain can also interact with actin remodelling proteins and modulate cytoskeletal contractility. Pseudocleavage furrow extension in the syncytial division cycles of Drosophila embryos requires the homeostatic control of conserved processes that control plasma membrane tension and actin contractility. We find that Syndapin plays an important role in promoting pseudocleavage furrow extension. We propose a model involving roles for Syndapin in membrane dynamics and direct or indirect effect on the cytoskeleton to explain how it affects pseudocleavage furrow growth, independent of its role in endocytosis.

Coordinated membrane and cytoskeletal remodeling activities are required for membrane extension in processes such as cytokinesis and syncytial nuclear division cycles in Drosophila. Pseudocleavage furrow membranes in the syncytial Drosophila blastoderm embryo show rapid extension and retraction regulated by actin-remodeling proteins. The F-Bar domain protein, Syndapin (Synd) is involved in membrane tubulation, endocytosis and uniquely in F-actin stability. Here we report a role for Synd in actin-regulated pseudocleavage furrow formation. Synd localized to these furrows and its loss resulted in short, disorganized furrows. Synd presence was important for the recruitment of the septin Peanut and distribution of Diaphanous and F-actin at furrows. Synd and Peanut were both absent in furrow initiation mutants of RhoGEF2 and Diaphanous, and in furrow progression mutants of Anillin. Synd overexpression in rhogef2 mutants reversed its furrow extension phenotypes, Peanut and Diaphanous recruitment and F-actin organization. We conclude that Synd plays an important role in pseudocleavage furrow extension and this role is also likely to be crucial in cleavage furrow formation during cell division.