Characterization of endothelin receptors in the inner medullary collecting duct of the rat.
Endothelins may be important regulators of renal inner medullary collecting duct (IMCD) function. These peptides are secreted in large amounts by IMCD cells and can, in turn, potently inhibit sodium and water transport systems in the IMCD. This study characterized endothelin (ET) receptors in the IMCD in order to gain insight into this unique renal autocrine system. Radioligand binding studies with 125I-ET-1 yielded a B(max) of 205.7 fmol/mg and a KD of 218 pM for ET-1. Similar studies with 125I-ET-3 yielded two populations of receptors for ET-3, one with a KD of 50 pM and one with a KD of 920 pM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of IMCD cells covalently labeling with 125I-ET-1 yielded two bands, one at 97 kDa with affinities of ET-1 greater than ET-2 much greater than ET-3 and one at 47 kDa with affinities ET-1 greater than or equal to ET-2 = ET-3. Reverse transcription and polymerase chain reaction revealed the presence of both endothelin receptor types A and B. These data indicate that IMCD cells have high affinity, high density receptors for endothelin and express both known types of endothelin receptor.
Kohan
D E
DE
Veterans Administration Medical Center, Salt Lake City, Utah.
Hughes
A K
AK
Perkins
S L
SL
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D016232:Endothelins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D007679:Kidney Medulla / Q000166:cytology / Q000378:metabolism*; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017466:Receptors, Endothelin
D003432:Cross-Linking Reagents; D016232:Endothelins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D017466:Receptors, Endothelin