期刊
JOURNAL OF CELL SCIENCE
卷 121, 期 21, 页码 3649-3663出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.018648
关键词
Epithelial cell polarity; Cell shape; Topology; Surface surveillance; Morphogenesis; 3D cell culture; Volume; Force
类别
资金
- LMC (light microscopy center)
- WBL (study of applied statistics) at ETH-Zurich, Switzerland
- CO-ME
- SNF
- KTI
- Roche Research Foundation
- Novartis Foundation
- UBS
Epithelial cells assemble into three-dimensional aggregates to generate lumen-containing organ substructures. Cells therein contact the extracellular matrix with their basal surface, neighbouring cells with their contact surface and the lumen with their apical surface. We investigated the development of single MDCK cells into aggregates with lumen using quantitative live-cell imaging to identify morphogenetic rules for lumen formation. In two-cell aggregates, membrane insertion into the contact surface established a preapical patch (PAP) characterized by the presence of the apical marker gp135, microvilli and the absence of E-cadherin. This PAP originated from a compartment that had hallmarks of an apical recycling endosome, and matured through Brefeldin-A-sensitive membrane trafficking and the establishment of tight junctions around itself. As a result of the activity of water and ion channels, an optically resolvable lumen formed. Initially, this lumen enlarged without changes in aggregate volume or cell number but with decreasing cell volumes. Additionally, the ROCK1/2-myosin-II pathway counteracted PAP and lumen formation. Thus, lumen formation results from PAP establishment, PAP maturation, lumen initiation and lumen enlargement. These phases correlate with distinct cell surface and volume patterns, which suggests that such morphometric parameters are regulated by trafficking, ROCK-mediated contractility and hydrostatic pressure or vice versa.
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