Endothelin A receptor-like immunoreactivity on the basal infoldings of rat renal tubules and collecting ducts

Abstract

We investigated the distribution of endothelin A (ET_A) receptor-like immunoreactivity in the rat kidney using affinity-purified antibodies against amino acid residues 403-417 of the rat ET_A receptor modified by the multiple antigen peptide complex system. Western blot analysis using the affinity-purified anti-ET_A antibody detected bands of approximately 47.3 and 64.5 kDa in the rat kidney. By light microscopy, ET_A receptor-like immunoreactivity was seen in the basal side of the renal tubules and collecting ducts. The most intense immunoreactivity was present in the distal renal tubules and inner medullary collecting ducts. In addition to the basal infoldings, immunoreactive puncta were scattered in the epithelial cells of the renal tubules and collecting ducts. Specimens prepared using the pre-embedding method were examined by electron microscopy, and some immunopositive signals were seen on the basal infodings of the renal tubules and collecting ducts. The lengths of immunopositive cytoplasmic membrane were far longer in the distal tubules and inner medullary collecting ducts than in the proximal tubules and outer medullary collecting ducts. Immunopositive signals were also sometimes observed in the thick portion of Henle's loop, but never in the thin portion. We have not previously detected immunopositive signals on the renal vascular systems with the antibody used here. These results suggest that endothelin acts on the basal infoldings through the ET_A receptor, particularly in the distal tubules and inner medullary collecting ducts, although involvement of the ET_B receptor cannot be excluded.