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

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

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...

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Bibliographic Details
Published in:The Journal of cell biology 2018-09, Vol.217 (9), p.3301-3311
Main Authors: Dambournet, Daphné, Sochacki, Kem A, Cheng, Aaron T, Akamatsu, Matthew, Taraska, Justin W, Hockemeyer, Dirk, Drubin, David G
Format: Article
Language:eng
Subjects:
Actin
Cells
Cells (biology)
Clathrin
Cytology
Dependence
Differentiation
Dynamic structural analysis
Embryo cells
Endocytosis
Fibroblasts
Genomes
Morphology
Neural stem cells
Platinum
Progenitor cells
Properties (attributes)
Proteins
Quantitative analysis
Stem cells
Studies
Transmission electron microscopy
Ultrastructure
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Summary: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 AP2μ2 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 AP2μ2 expression regulation.
ISSN:0021-9525
1540-8140