Comparison of Endothelin-A and Endothelin-B Receptor Distribution Visualized by Radioligand Binding versus Immunocytochemical Localization using Subtype Selective Antisera

Abstract

Molecular studies have predicted the existence in human tissue of splice variants and modifications to the amino acid sequence of endothelin receptors that may modulate function. Endothelin-A (ETA) receptors were visualized by ligand binding and autoradiography to the renal vasculature throughout the cortex and medulla, including the large arcuate arteries, adjacent veins and arterioles. Lower binding densities were visualized to the vasa recta and glomeruli. A similar pattern of staining was revealed by ETA selective antisera, with the higher resolution demonstrating ETA receptors confined to smooth muscle cells. Staining was also detected to the vasa recta and glomeruli. Ligand binding revealed a more heterogeneous endothelin-B (ETB) receptor distribution with high densities concentrated in the medulla. Three different sitedirected ETB antisera demonstrated a similar pattern of staining to the endothelium lining all renal vessels but not to the smooth muscle. Staining was also detected to glomerular endothelial cells as well as epithelial cells lining the renal tubule, particularly the collecting ducts, consistent with high binding densities observed in the medulla by autoradiography. There was no evidence for a differential distribution in either ETA or ETB receptors visualized by the two techniques that might have indicated modified receptors or further subtypes in the human kidney.