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Brain Res. 1987 Sep 8;420(1):32-8.

Blood-brain barrier transcytosis of insulin in developing rabbits.

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  • 1Department of Neurosurgery, UCLA School of Medicine 90024.

Abstract

Previous studies with isolated brain microvessels have suggested that blood insulin is selectively transported through the brain capillary, i.e. the blood-brain barrier (BBB), by receptor-mediated transcytosis. The purpose of the present study is to demonstrate in vivo the uptake of circulating 125I-insulin by brain using thaw-mount autoradiography. However, metabolism of systemic 125I-insulin to 125I-tyrosine would allow for brain uptake of 125I-tyrosine and this would preclude interpretation of the autoradiogram. Therefore, the present studies were performed in developing rabbits, since plasma protein degradation of peptides is greatly reduced in developing animals. 125I-insulin was infused via the carotid artery at a rate of 0.25 ml/min for 1, 5, or 10 min, and the mean brain uptake, relative to a [3H]albumin reference, was 99.3 +/- 5.5%, 110.1 +/- 4.3%, and 143.6 +/- 7.9%, respectively. This uptake was saturable by simultaneously infusing unlabeled insulin. Thaw-mount autoradiography of rabbit brain after a 10-min infusion of 125I-insulin revealed silver grains in the pericapillary space and well within the brain parenchyma. HPLC analysis of acid-ethanol extracts of rabbit blood after a 10-min infusion showed virtually all of the 125I-radioactivity co-migrated with a known insulin standard on a reverse-phase column, indicating minimal degradation of infused 125I-insulin. HPLC analysis of brain radioactivity showed the major peak co-migrated with 125I-insulin and this peak was precipitated by an anti-insulin antiserum. The correlation of the transport data, the autoradiography, and the HPLC analysis support the model that brain insulin originates from blood via receptor-mediated transport of the peptide at the BBB.

PMID:
3315116
[PubMed - indexed for MEDLINE]
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