Hyperosmotic urea reversibly opens the tight junctions between brain capillary endothelial cells in cell culture

The effect of hyperosmotic solutions of urea on primary cultures of bovine brain microvessel endothelial cells was examined. Confluent monolayers of cells positive for Factor VIII-related antigen were obtained by seven days in culture. The cells were: (a) incubated in media containing 1 M, 2 M or 3 M urea and horseradish peroxidase (HRP) for various periods of time and then examined by transmission electron microscopy. (b) Exposed to hypertonic urea solutions and ¹⁴C-3-0-methyl-D-glucose for determination of the intracellular water space. In control cultures endothelial cells were bound together by tight junctional complexes over 91% of which excluded HRP. In experimental cultures 82% of the interendothelial clefts became permeable to HRP after one minute of incubation with 3 M urea. The degree of cell shrinkage corresponded well with the extent of junctional opening. In monolayers examined 24 hours following removal of urea from the media more than 63% of the intercellular clefts were impermeable to HRP. These observations indicate that hyperosmotic solutions of urea reversibly open the tight junctions between brain microvessel endothelial cells in tissue culture. Decrease in cell volume appears to be linked to the increased junctional permeability.