[ 125 I‐ His 9 ]‐Ghrelin, a novel radioligand for localizing GHS orphan receptors in human and rat tissue; up‐regulation of receptors with atherosclerosis

Ghrelin is the recently identified endogenous ligand for the cloned growth hormone secretagogue receptor (GHS‐R). We have characterized for the first time the binding of human [ 125 I‐ His 9 ]‐ghrelin to normal human and rat tissue and demonstrated expression of this ‘orphan’ receptor that has previously been predicted to exist from mRNA. Furthermore, we have discovered that [ 125 I‐ His 9 ]‐ghrelin density is significantly increased in atherosclerosis. [ 125 I‐ His 9 ]‐Ghrelin bound to non‐diseased human heart (left ventricle) with an association rate constant (k obs ) of 0.16±0.004 min −1 , a dissociation rate constant of 0.068±0.0005 min −1 (kinetically derived K D of 0.1 n M ; n =5 individuals±s.e.mean), a K D of 0.43±0.08 n M and B max of 7.8±0.9 fmol mg −1 protein ( n =6 individual±s.e.mean). Specific [ 125 I‐ His 9 ]‐ghrelin binding was to the human vasculature including aorta, coronary, pulmonary, arcuate arteries in the kidney and saphenous veins. In rat tissues, binding sites were also localized to the vasculature in peripheral tissues as well as the granular layer of the cerebellum in the CNS. [ 125 I‐ His 9 ]‐Ghrelin binding was significantly up‐regulated (3 – 4 fold) in both atherosclerotic coronary arteries and saphenous vein grafts with advanced intimal thickening, compared with normal vessels ( P <0.05). Our results suggest that the native receptor for [ 125 I‐ His 9 ]‐ghrelin may be widely distributed in the human cardiovascular system. Furthermore, changes in the density of this proposed ghrelin receptor implicates this new transmitter system in the development of atherosclerosis and may therefore represent a novel therapeutic target in the treatment of cardiovascular disease.British Journal of Pharmacology (2001) 134 , 143–149; doi: 10.1038/sj.bjp.0704228