Journal
JOURNAL OF CELL SCIENCE
Volume 136, Issue 15, Pages -Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.260668
Keywords
Tight junctions; Barrier function; Epithelia; Live imaging
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Epithelial barrier function is commonly analyzed using transepithelial electrical resistance or traceable macromolecules to measure changes in global barrier function. However, these methods lack sensitivity to detect local or transient barrier breaches and do not reveal the location of barrier leaks. To overcome these limitations, the zinc-based ultrasensitive microscopic barrier assay (ZnUMBA) was previously developed, which allows for detection of local tight junction leaks with high spatiotemporal resolution.
Epithelial barrier function is commonly analyzed using transepithelial electrical resistance, which measures ion flux across a monolayer, or by adding traceable macromolecules and monitoring their passage across the monolayer. Although these methods measure changes in global barrier function, they lack the sensitivity needed to detect local or transient barrier breaches, and they do not reveal the location of barrier leaks. Therefore, we previously developed a method that we named the zinc-based ultrasensitive microscopic barrier assay (ZnUMBA), which overcomes these limitations, allowing for detection of local tight junction leaks with high spatiotemporal resolution. Here, we present expanded applications for ZnUMBA. ZnUMBA can be used in Xenopus embryos to measure the dynamics of barrier restoration and actin accumulation following laser injury. ZnUMBA can also be effectively utilized in developing zebrafish embryos as well as cultured monolayers of Madin-Darby canine kidney (MDCK) II epithelial cells. ZnUMBA is a powerful and flexible method that, with minimal optimization, can be applied to multiple systems to measure dynamic changes in barrier function with spatiotemporal precision.
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