A system for whole-blood perfusion of the bovine eye through the cilioretinal artery was developed and the distribution and binding of 125I-labelled insulin and human growth hormone (HGH) were studied autoradiographically. Sodium fluorescein was used as a tracer to monitor blood retinal barrier integrity, and electron microscopy was used to determine structural preservation after perfusion fixation. With this system, barrier integrity and structural perservation of both the neural retina and the retinal blood vessels were regularly obtained, with perfusion periods of as long as 5 hours. By quantitative light microscopy autoradiography, insulin binding sites were identified on the endothelial cells of retinal capillaries after perfusion with blood containing 125I-insulin. 125I-insulin binding was competitively inhibited by the addition of unlabelled insulin to the perfusing blood. By contrast the low level of binding of HGH to retinal capillaries was nonspecific. Electron microscopy autoradiography revealed 125I-insulin autoradiographic grains lying over the endothelial cell wall, over pinocytotic vesicles, and over the cytoplasm of both endothelial cells and pericytes. This suggests that, after binding to the cell surface, some insulin passes into the cell cytoplasm. However, neither 125I-HGH penetrated as far as the retina in the periods studied.
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