Sub-type selective ligands revealed a differential distribution of endothelin (ET) receptors within human adrenal glands. High densities of ETA receptors were localized, using [125I]-PD151242, to the smooth muscle layer of the arteries, smaller vessels within the capsular plexus and to the secretory cells of zona glomerulosa (KD = 139.8 +/- 39.7, Bmax = 69.7 +/- 9.1 fmol mg-1 protein, mean of 3 individuals+/-sem). ETB receptors were present in the medulla (KD = 145.2 +/- 16.4, Bmax = 75.5 +/- 12.3), zona glomerulosa (KD = 100.6 +/- 35.1, Bmax = 63.1 +/- 10.0), fasiculata (KD 145.1 +/- 16.2, Bmax = 67.9 +/- 6.9) and reticularis (KD = 118.2 +/- 18.6, Bmax = 71.9 +/- 6.5). ETB receptors were not detected within the smooth muscle of the vasculature. Messenger RNA encoding both sub-types was present in adrenals. ET-like immunoreactivity was localized to the cytoplasm of the endothelial cells from arteries supplying the gland and resistance vessels within the capsular plexus. Staining was also detected in these cells using anti-big ET-1 and less intensely with anti-big ET-2 antisera but not within cells within the cortex or medulla. Big ET-3-like immunoreactivity was localized to secretory cells of the medulla. Staining was not found using antiserum that could detect ET-3, suggesting further processing of big ET-3 may occur within the plasma, and that the adrenals could be a source of ET-3. The presence of ET-1 was confirmed by high performance liquid chromatography and radioimmunoassay although ET-3 was not detected. The results suggest that ET-1 is the predominant mature isoform, which is localized mainly to adrenal vasculature, particularly the capsular plexus, and may contribute to blood flow regulation in the gland.