Genome-edited human stem cells expressing fluorescently labeled endocytic markers allow quantitative analysis of clathrin-mediated endocytosis during differentiation

被引：44

作者：

Dambournet, Daphne ^{[1
]}

Sochacki, Kem A. ^{[2
]}

Cheng, Aaron T. ^{[3
]}

Akamatsu, Matthew ^{[1
]}

Taraska, Justin W. ^{[2
]}

Hockemeyer, Dirk ^{[1
]}

Drubin, David G. ^{[1
]}

机构：

[1] Univ Calif Berkeley, Dept Mol & Cell Biol, 229 Stanley Hall, Berkeley, CA 94720 USA

[2] NHLBI, NIH, Bldg 10, Bethesda, MD 20892 USA

[3] GlaxoSmithKline, Collegeville, PA USA

来源：

JOURNAL OF CELL BIOLOGY | 2018年 / 217卷 / 09期

基金：

美国国家卫生研究院;

关键词：

VESICLE FORMATION; MAMMALIAN-CELLS; COATED PITS; ACTIN CYTOSKELETON; MEMBRANE; DYNAMICS; SPECIFICATION; TRAFFICKING; FIBROBLASTS; LATTICES;

D O I：

10.1083/jcb.201710084

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We developed a general approach for investigation of how cellular processes become adapted for specific cell types during differentiation. Previous studies reported substantial differences in the morphology and dynamics of clathrin-mediated endocytosis (CME) sites. However, associating specific CME properties with distinct differentiated cell types and determining how these properties are developmentally specified during differentiation have been elusive. Using genome-edited human embryonic stem cells, and isogenic fibroblasts and neuronal progenitor cells derived from them, we established by live-cell imaging and platinum replica transmission electron microscopy that CME site dynamics and ultrastructure on the plasma membrane are precisely reprogrammed during differentiation. Expression levels for the endocytic adaptor protein AP2 mu 2 were found to underlie dramatic changes in CME dynamics and structure. Additionally, CME dependency on actin assembly and phosphoinositide-3 kinase activity are distinct for each cell type. Collectively, our results demonstrate that key CME properties are reprogrammed during differentiation at least in part through AP2 mu 2 expression regulation.

引用

页码：3301 / 3311

页数：11