A noninvasive procedure for in vivo assay of a lung amine endothelial receptor

Abstract

Lung endothelial N-isopropyl-p-iodoamphetamine (IMP) binding sites were assessed applying principles of competitive binding assay adapted for in vivo measurements obtained by digital imaging. Data were acquired following the method published by Rahimian et al., a modification of the dual indicator dilution technique of Chinard and Crone. Iodine-123 (123I) IMP, the test cellular tracer, and technetium-99m (99mTc) dextran, the reference vascular tracer were imaged during their first pass through the superior vena cava, right heart, lungs, and left heart in West African dwarf goats. The lung fractional extraction of IMP diminished progressively from 0.96 to 0.20 as the amount of IMP in the test tracer boluses was gradually increased from 0.6 to 150 mg. This demonstrated that lung extraction of IMP is by way of a saturable binding system, presumably receptors. The dissociation constant of IMP-lung binding sites reaction was calculated by Scatchard plot and found to be 11.7 mg. The amount of IMP bound at saturation (R), was found to be 30 mg. Assuming that a single molecule of IMP bound a single receptor, the total number of free receptors was computed as the Avogadro's number times R, divided by the IMP molecular weight, and found to be 6.04 X 10(19). Using a computer model, it was determined that the 20 mg per bolus isotherm was the most sensitive for measuring the number of total free receptors (binding sites). This is the first time, to our knowledge, that noninvasive in vivo assessment of receptors in lung has been accomplished. Basically, the method used can be applied in humans and, also, to assess receptors in organs other than the lungs.