Purposes: First, to develop an improved retinal capillary endothelial cell culture system which exhibits some of the physiologic features of the blood- retinal barrier; second, to use this model to determine how histamine and chemical conditions of diabetes effect expression of the tight junction protein, ZO-1; and third, to discuss application of the Henle-Koch postulates to the problem of diabetic retinopathy. Methods: Bovine retinal capillary endothelial cells were exposed to varying serum and growth factor concentrations, as well as astrocyte-conditioned medium, in order to establish a model of the blood-retinal barrier. Cells were also exposed to varying concentrations of histamine, and of insulin and glucose. The expression of ZO-1 tight junction protein was determined by immunocytochemistry and immunoblotting. Results: Modified concentrations of growth factors reduced endothelial cell proliferation, without reducing viability. Astrocyte conditioned medium increased ZO-1 protein content. Histamine reduced ZO-1 protein content. Both high glucose (20mM) and low insulin (10-12M) reduced ZO-1 protein content compared to control conditions (5mM glucose and 10-9 M insulin). Conclusions: Control of culture conditions results in a more physiologic in vitro model of the blood- retinal barrier. Soluble factors from astrocytes promote tight junction formation. Both histamine anti chemical conditions of diabetes diminish tight junction formation. These factors may mediate blood-retinal barrier breakdown in diabetic retinopathy. Henle-Koch postulates for diabetic retinopathy are presented.
Gardner, T. W. (1995). Histamine, ZO-1 and increased blood-retinal barrier permeability in diabetic retinopathy. In Transactions of the American Ophthalmological Society (Vol. 93, pp. 583–621). https://doi.org/10.1016/s0002-9394(14)70570-7