Insulin binding to microvascular endothelium of intact heart: a kinetic and morphometric analysis

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Abstract
We have studied insulin binding to blood vessels of the intact, beating heart. The beating hearts were perfused with [125I]iodoinsulin (3 X 10(-10) M) followed by perfusion with unlabeled insulin. The unlabeled insulin displaced the bound [125I]iodoinsulin in direct proportion to the concentration of the unlabeled hormone. Perfusion with unlabeled insulin at 10(-11) M elicited a significant displacement of [125I]iodoinsulin, with a maximal effect at 10(-6) M. Unlabeled proinsulin also displaced [125I]iodoinsulin in a dose-dependent manner, being 1% as potent as insulin. Perfusion with unrelated peptides had no effect. Radioautographic counting of 125I grains indicated that greater than 95% of the grain counts over blood vessels were within the microvessels. When [125I]iodoinsulin perfusion was followed by perfusion with unlabeled insulin at 10(-6) M, there was a 50% decrease in grain counts over the microvessels (versus perfusion with [125I]iodoinsulin alone); with coperfusion of [125I]iodoinsulin and 10(-6) M unlabeled insulin, an 80% decrease in grain counts occurred. Electron microscopic radioautography indicated that the 125I grains were associated with the vascular endothelial cells. We conclude that specific insulin receptors are present on endothelial cells of microvessels in the intact heart.