Syntaxin 16 Regulates Lumen Formation during Epithelial Morphogenesis
- Authors
- Jung, J.J., Inamdar, S.M., Tiwari, A., Ye, D., Lin, F., and Choudhury, A.
- ID
- ZDB-PUB-130604-15
- Date
- 2013
- Source
- PLoS One 8(4): e61857 (Journal)
- Registered Authors
- Lin, Fang, Ye, Ding
- Keywords
- none
- MeSH Terms
-
- Animals
- Cadherins/genetics
- Cadherins/metabolism
- Cell Count
- Cell Membrane/metabolism
- Cell Membrane/ultrastructure
- Collagen
- Dogs
- Drug Combinations
- Embryo, Nonmammalian
- Endosomes/metabolism
- Endosomes/ultrastructure
- Gene Expression Regulation, Developmental*
- Golgi Apparatus/metabolism
- Golgi Apparatus/ultrastructure
- Intercellular Junctions/genetics
- Intercellular Junctions/metabolism*
- Intercellular Junctions/ultrastructure
- Kidney/growth & development
- Kidney/metabolism*
- Kidney/ultrastructure
- Laminin
- Madin Darby Canine Kidney Cells
- Protein Transport
- Proteoglycans
- SNARE Proteins/genetics
- SNARE Proteins/metabolism
- Signal Transduction
- Spindle Apparatus/metabolism
- Spindle Apparatus/ultrastructure
- Syntaxin 16/genetics
- Syntaxin 16/metabolism*
- Transgenes
- Zebrafish
- PubMed
- 23626741 Full text @ PLoS One
The formation and maintenance of cell-cell junctions, both under physiological and pathological conditions, requires the targeting and trafficking of junctional proteins. Proteins of the syntaxin (Stx)-family localize to a variety of subcellular membranes and contribute to intracellular transport of cargo by regulating vesicle fusion events at these sites. Unlike plasma membrane localized Stxs, the roles of endosome- and Golgi-localized stx proteins in epithelial morphogenesis are less understood. Here we show that Stx16– an endosome- and Golgi-localized target-membrane soluble N-ethylmaleimide attachment protein receptor (t-SNARE) that plays a role in membrane trafficking between these compartments – is essential for lumen development. In cultured Madin Darby Canine Kidney (MDCK) cells, Stx16 was selectively upregulated as sparsely plated cells attained confluency. Stx16-depleted confluent monolayers consistently showed lower transepithelial resistance than control monolayers, and failed to maintain endogenous and ectopically expressed E-cadherin at the adherens junctions due to decreased recycling. We further found that whereas cysts formed by MDCK cells cultured in Matrigel have a single hollow lumen, those formed by stx16-depleted counterparts had multiple lumens, due to abnormal orientiation of the mitotic spindle. Finally, a similar role for stx16 function in vivo is indicated by our analysis of pronephric-duct development in zebrafish expressing the claudinB:lynGFP transgene; lack of stx16 function in this structure (in stx16-morphant embryos) led to the development of enlarged, torturous pronephric ducts with more than one lumen. Taken together, our in vitro and in vivo studies establish a role for Stx16 in maintaining the integrity of cell-cell junctions, and thereby in morphogenesis of the kidney epithelial lumen.