Asymmetric formation of coated pits on dorsal and ventral surfaces at the leading edges of motile cells and on protrusions of immobile cells

被引:29
作者
Kural, Comert [1 ,2 ,3 ]
Akatay, Ahmet Ata [4 ]
Gaudin, Raphael [1 ,2 ]
Chen, Bi-Chang [5 ]
Legant, Wesley R. [5 ]
Betzig, Eric [5 ]
Kirchhausen, Tom [1 ,2 ,6 ]
机构
[1] Harvard Univ, Sch Med, Dept Cell Biol, Boston, MA 02115 USA
[2] Boston Childrens Hosp, Program Cellular & Mol Med, Boston, MA 02115 USA
[3] Ohio State Univ, Dept Phys, Columbus, OH 43210 USA
[4] Ohio State Univ, Biophys Grad Program, Columbus, OH 43210 USA
[5] Howard Hughes Med Inst, Ashburn, VA 20147 USA
[6] Harvard Univ, Sch Med, Dept Pediat, Boston, MA 02115 USA
基金
美国国家卫生研究院;
关键词
CLATHRIN-MEDIATED ENDOCYTOSIS; PLASMA-MEMBRANE AREA; DYNAMIN RECRUITMENT; GROWTH CONE; FIBROBLASTS; MOVEMENTS; ACTIN; LOCALIZATION; LOCOMOTION; RECEPTORS;
D O I
10.1091/mbc.E15-01-0055
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Clathrin/AP2-coated vesicles are the principal endocytic carriers originating at the plasma membrane. In the experiments reported here, we used spinning-disk confocal and lattice light-sheet microscopy to study the assembly dynamics of coated pits on the dorsal and ventral membranes of migrating U373 glioblastoma cells stably expressing AP2 tagged with enhanced green fluorescence (AP2-EGFP) and on lateral protrusions from immobile SUM159 breast carcinoma cells, gene-edited to express AP2-EGFP. On U373 cells, coated pits initiated on the dorsal membrane at the front of the lamellipodium and at the approximate boundary between the lamellipodium and lamella and continued to grow as they were swept back toward the cell body; coated pits were absent from the corresponding ventral membrane. We observed a similar dorsal/ventral asymmetry on membrane protrusions from SUM159 cells. Stationary coated pits formed and budded on the remainder of the dorsal and ventral surfaces of both types of cells. These observations support a previously proposed model that invokes net membrane deposition at the leading edge due to an imbalance between the endocytic and exocytic membrane flow at the front of a migrating cell.
引用
收藏
页码:2044 / 2053
页数:10
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