ABSTRACT
The actin cytoskeleton shapes cells and also organizes internal membranous compartments. In particular, it interacts with membranes for intracellular transport of material in mammalian cells, yeast, or plant cells. Tubular membrane intermediates, pulled along microtubule tracks, are formed during this process and destabilize into vesicles. While the role of actin in tubule destabilization through scission is suggested, literature also provides examples of actin-mediated stabilization of membranous structures. To directly address this apparent contradiction, we mimic the geometry of tubular intermediates with preformed membrane tubes. The growth of an actin sleeve at the tube surface is monitored spatiotemporally. Depending on network cohesiveness, actin is able to entirely stabilize or locally maintain membrane tubes under pulling. On a single tube, thicker portions correlate with the presence of actin. These structures relax over several minutes and may provide enough time and curvature geometries for other proteins to act on tube stability.
ABSTRACT
Cell and tissue mechanical properties are paramount in controlling morphogenesis. Microaspiration techniques allow measuring the absolute values of mechanical properties in space and time in vivo. Here, we explain how to build a microaspiration setup that can be used for both cellular and tissue scale measurements. At the cellular scale, microaspiration allows the mapping in space and time of surface tensions of individual interfaces within a tissue to understand the forces shaping it. At the tissue scale, microaspiration can be used to measure macroscopic mechanical properties such as the viscoelasticity and tissue surface tension that regulate the dynamics of tissue deformation. Based on a simple and cost-effective apparatus, these two complementary microaspiration techniques provide unique tools for exploring cell and tissue mechanics in vivo.
