Dynamics of tight junctions: GFP chimera and confocal laser scanning microscopy

Tight junctions (TJ) separate distinct components of the apical and basolateral cell surface domains and at the same time act as a dynamic gate to regulate passage of molecules through the paracellular pathway. Understanding TJ regulation processes can significantly help to improve drug delivery while preventing damage to the epithelial barrier. Confocal laser scanning microscopy (CLSM) is used in conjunction with transepithelial electrical resistance and paracellular flux measurements as well as protein analysis tools to study the TJ characteristics under various conditions. Fusion constructs (chimera) between Green Fluorescent Protein (GFP) and either occludin or ZO-1, two of the major TJ proteins, were made and stably expressed in Madin Darby Canine kidney (MDCK) cells. With this approach information is obtained on the formation, opening and reformation of TJ in living cells under physiological conditions as well as under the influence of drugs and permeation enhancers.

ETH Research Database: id 8915

Contacts: Prof. Dr. Heidi Wunderli-Allenspach, Dr. Stefanie Krämer