PUBLICATION
Zinc-based Ultrasensitive Microscopic Barrier Assay (ZnUMBA): a live imaging method to detect local barrier breaches
- Authors
- Higashi, T., Stephenson, R.E., Schwayer, C., Huljev, K., Higashi, A.Y., Heisenberg, C.P., Chiba, H., Miller, A.L.
- ID
- ZDB-PUB-230718-41
- Date
- 2023
- Source
- Journal of Cell Science 136(15): (Journal)
- Registered Authors
- Heisenberg, Carl-Philipp
- Keywords
- Barrier function, Epithelia, Live imaging, Tight junctions
- MeSH Terms
-
- Actins
- Animals
- Dogs
- Epithelial Cells*
- Madin Darby Canine Kidney Cells
- Tight Junctions
- Zebrafish
- Zinc*
- PubMed
- 37461809 Full text @ J. Cell Sci.
Citation
Higashi, T., Stephenson, R.E., Schwayer, C., Huljev, K., Higashi, A.Y., Heisenberg, C.P., Chiba, H., Miller, A.L. (2023) Zinc-based Ultrasensitive Microscopic Barrier Assay (ZnUMBA): a live imaging method to detect local barrier breaches. Journal of Cell Science. 136(15).
Abstract
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. While 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 developed a method that we named Zinc-based Ultrasensitive Microscopic Barrier Assay (ZnUMBA), which overcomes these limitations, allowing for detection of local tight junction leaks with high spatiotemporal resolution (Chumki et al., 2022; Higashi et al., 2023; Stephenson et al., 2019; Varadarajan et al., 2022). 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 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping