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Syntaxin 16 regulates lumen formation during epithelial morphogenesis
Journal article   Open access   Peer reviewed

Syntaxin 16 regulates lumen formation during epithelial morphogenesis

Jae-Joon Jung, Shivangi M Inamdar, Ajit Tiwari, Ding Ye, Fang Lin and Amit Choudhury
PloS one, Vol.8(4), pp.e61857-e61857
2013
DOI: 10.1371/journal.pone.0061857
PMCID: PMC3633931
PMID: 23626741
url
https://doi.org/10.1371/journal.pone.0061857View
Published (Version of record) Open Access

Abstract

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.
Cadherins - metabolism SNARE Proteins - genetics Cell Count Endosomes - metabolism Kidney - metabolism Laminin Spindle Apparatus - metabolism Syntaxin 16 - genetics Gene Expression Regulation, Developmental Endosomes - ultrastructure Madin Darby Canine Kidney Cells Cell Membrane - metabolism Cadherins - genetics Spindle Apparatus - ultrastructure Syntaxin 16 - metabolism Transgenes Golgi Apparatus - ultrastructure Proteoglycans Signal Transduction Intercellular Junctions - metabolism Kidney - growth & development Zebrafish Cell Membrane - ultrastructure Intercellular Junctions - genetics Kidney - ultrastructure Protein Transport Animals Collagen Dogs Embryo, Nonmammalian Golgi Apparatus - metabolism Intercellular Junctions - ultrastructure SNARE Proteins - metabolism Drug Combinations

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