Actin-based protrusions of migrating neutrophils are intrinsically lamellar and facilitate direction changes

eLife

Eric Betzig¹  Susan M Nicholson-Dykstra²  Henry Higgs²  Megan Riel-Mehan³  Graham T Johnson³  Samuel J Lord⁴  Lillian K Fritz-Laylin⁴  Thomas E Ferrin⁵  Thomas D Goddard⁵  R Dyche Mullins⁶  Bi-Chang Chen⁶ 

[1] Animated Cell, Allen Institute for Cell Science, Seattle, United States;Department of Biochemistry and Cell Biology, Dartmouth Geisel School of Medicine, Hanover, United States;Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, United States;Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States;Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States;Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States;

关键词: cell migration;    microscopy;    visualization;    actin;    arp2/3;    amoeboid motility;   

DOI  :  10.7554/eLife.26990

来源: DOAJ

【 摘 要 】

Leukocytes and other amoeboid cells change shape as they move, forming highly dynamic, actin-filled pseudopods. Although we understand much about the architecture and dynamics of thin lamellipodia made by slow-moving cells on flat surfaces, conventional light microscopy lacks the spatial and temporal resolution required to track complex pseudopods of cells moving in three dimensions. We therefore employed lattice light sheet microscopy to perform three-dimensional, time-lapse imaging of neutrophil-like HL-60 cells crawling through collagen matrices. To analyze three-dimensional pseudopods we: (i) developed fluorescent probe combinations that distinguish cortical actin from dynamic, pseudopod-forming actin networks, and (ii) adapted molecular visualization tools from structural biology to render and analyze complex cell surfaces. Surprisingly, three-dimensional pseudopods turn out to be composed of thin (<0.75 µm), flat sheets that sometimes interleave to form rosettes. Their laminar nature is not templated by an external surface, but likely reflects a linear arrangement of regulatory molecules. Although we find that Arp2/3-dependent pseudopods are dispensable for three-dimensional locomotion, their elimination dramatically decreases the frequency of cell turning, and pseudopod dynamics increase when cells change direction, highlighting the important role pseudopods play in pathfinding.

【 授权许可】

Unknown