Endothelin-A and -B receptors, superoxide, and Ca2+ signaling in afferent arterioles.
It is unknown if endothelin-A and -B receptors (ET(A)R and ET(B)R) activate the production of superoxide via NAD(P)H oxidase and subsequently stimulate the formation of cyclic adenine diphosphate ribose (cADPR) in afferent arterioles. Vessels were isolated from rat kidney and loaded with fura 2. Endothelin-1 (ET-1) rapidly increased cytosolic Ca(2+) concentration ([Ca(2+)](i)) by 303 nM. The superoxide dismutase mimetic tempol, the NAD(P)H oxidase inhibitor apocynin, and nicotinamide, an inhibitor of ADPR cyclase, diminished the response by approximately 60%. The ET(B)R agonist sarafotoxin 6c (S6c) increased peak [Ca(2+)](i) by 117 nM. Subsequent addition of ET-1 in the continued presence of S6c caused an additional [Ca(2+)](i) peak of 225 nM. Neither nicotinamide or 8-bromo- (8-Br) cADPR nor apocynin decreased the [Ca(2+)](i) response to S6c, but inhibited the subsequent [Ca(2+)](i) response to ET-1. The ET(B)R blockers BQ-788 and A-192621 prevented the S6c [Ca(2+)](i) peak and reduced the ET-1 response by more than one-half, suggesting an ET(B)R/ET(A)R interaction. In contrast, the ET(A)R blocker BQ-123 had no effect on the S6c [Ca(2+)](i) peak and obliterated the subsequent ET-1 response. ET-1 immediately stimulated superoxide formation (measured with TEMPO-9-AC, 68 arbitrary units) that was inhibited 95% by apocynin or diphenyl iodonium. S6c or IRL-1620 increased superoxide by 8% of that caused by subsequent ET-1 addition. We conclude that ET(A)R activation of afferent arterioles increases the formation of superoxide that accounts for approximately 60% of subsequent Ca(2+) signaling. ET(B)R activation appears to result in only minor increases in superoxide production. Nicotinamide and 8-Br-cADPR results suggest that ET-1 (and primarily ET(A)R) causes the activation of vascular smooth muscle cell-ADPR cyclase.
Fellner
Susan K
SK
Dept. of Cell and Molecular Physiology, Univ. of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7545, USA. sfellner@med.unc.edu
Arendshorst
William
W
D036541:ADP-ribosyl Cyclase / Q000037:antagonists & inhibitors; D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D001160:Arterioles / Q000502:physiology; D001713:Biphenyl Compounds / Q000494:pharmacology; D001920:Bradykinin / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D003497:Cyclic N-Oxides / Q000494:pharmacology; D042783:Endothelial Cells / Q000502:physiology; D065131:Endothelin B Receptor Antagonists; D016232:Endothelins / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000502:physiology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009536:Niacinamide / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D009861:Onium Compounds / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D044022:Receptor, Endothelin A / Q000819:agonists / Q000378:metabolism*; D044023:Receptor, Endothelin B / Q000819:agonists / Q000378:metabolism*; D012079:Renal Circulation / Q000502:physiology*; D013113:Spin Labels; D013481:Superoxides / Q000378:metabolism*; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D014757:Viper Venoms / Q000494:pharmacology
C120508:A 192621; D000098:Acetophenones; D001713:Biphenyl Compounds; D003497:Cyclic N-Oxides; D065131:Endothelin B Receptor Antagonists; D016232:Endothelins; D004791:Enzyme Inhibitors; D009842:Oligopeptides; D009861:Onium Compounds; D010446:Peptide Fragments; D010880:Piperidines; D011759:Pyrrolidines; D044022:Receptor, Endothelin A; D044023:Receptor, Endothelin B; D013113:Spin Labels; D014662:Vasoconstrictor Agents; D014757:Viper Venoms; C057354:sarafotoxins s6; C031291:diphenyliodonium; D013481:Superoxides; C074308:sovateltide; D009536:Niacinamide; C086539:BQ 788; C056165:acetovanillone; D019255:NADPH Oxidases; D036541:ADP-ribosyl Cyclase; D001920:Bradykinin; C001803:tempol; D019331:NG-Nitroarginine Methyl Ester