Popp David, Yamamoto Akihiro, Iwasa Mitsusada, Maéda Yuichiro
ERATO Actin Filament Dynamics Project, JST, c/o RIKEN SPring-8 Center, Kouto, Sayo, Hyogo 679-5148, Japan.
Biochem Biophys Res Commun. 2006 Dec 15;351(2):348-53. doi: 10.1016/j.bbrc.2006.10.041. Epub 2006 Oct 16.
Various actin assemblies within the cell regulate many cellular processes such as cell shape and motility. The mechanical properties of these networks are challenging to measure in vivo. They have been studied in solution by indirect observation methods, such as multiple ball tracking. However, little is known about the behavior of such networks near the crowded cell membrane. Here we used in vitro TIRF microscopy to directly probe the formation of actin networks in real-time near a hydrophilic surface in the presence of crowding agents. We find that under these conditions actin does not form a mesh like network, but either textured nematic liquid crystals or a bundled network. We are directly able to follow the thermal fluctuations of actin filaments within these networks. Prearranged parallel networks of actin filaments near the crowded cell membrane could play a role in the rapid formation of stress fibers or microvilli.
细胞内的各种肌动蛋白组装体调节许多细胞过程,如细胞形状和运动性。这些网络的力学性质在体内测量具有挑战性。它们已通过间接观察方法在溶液中进行研究,例如多球跟踪。然而,对于此类网络在拥挤细胞膜附近的行为知之甚少。在这里,我们使用体外全内反射荧光显微镜(TIRF)在存在拥挤剂的情况下,实时直接探测亲水性表面附近肌动蛋白网络的形成。我们发现,在这些条件下,肌动蛋白不会形成类似网状的网络,而是形成纹理化的向列液晶或束状网络。我们能够直接跟踪这些网络中肌动蛋白丝的热涨落。拥挤细胞膜附近预先排列的平行肌动蛋白丝网络可能在应力纤维或微绒毛的快速形成中发挥作用。