Endothelin-1 activates phospholipases and channels at similar concentrations in porcine coronary arteries.
Sensitivity of endothelin-1 (ET-1)-ion channel interactions has been proposed to exceed that of ET-1-phospholipase activation in vascular smooth muscle. We wanted to determine whether short-circuiting ion channels with staphylococcal alpha-toxin pores would shift the ET-1-force relation to the right as predicted from the above proposal. Medium size porcine coronary arteries (outer diameter 0.7-1.5 mm) were mounted on isometric force transducers. ET-1 concentration response curves were compared between intact rings and those subjected to alpha-toxin treatment with Ca buffered at 0.1 microM. The EC50 for treated rings (1.5 +/- 1.0 nM, n = 5 pigs) was similar to that for intact rings (1.9 +/- 0.4 nM). The Ca sensitivity of the alpha-toxin-treated rings (EC50 = 0.43 +/- 0.08 microM) was similar to that reported by other laboratories for intact and alpha-toxin-treated arteries and was shifted eightfold to the left by a high concentration of ET-1 (10 nM). Measurements of [32P]phosphatidic acid ([32P]PA) levels were used to evaluate phospholipase activity in intact arteries. The time courses for [32P]PA production and contraction were similar in response to high (100 nM) and to low (1 nM) ET-1. Significant increases in both steady-state contraction and [32P]PA occurred over a wide range of ET-1 concentrations tested (0.3-100 nM). Our findings support the concept that ET-1-phospholipase coupling is operative over the whole concentration range that induces contractile responses. It is suggested that both Ca entry and Ca sensitization processes are activated by ET-1 at low concentrations (<EC50) and that both processes contribute significantly to the integrated response.
Jones
A W
AW
Department of Physiology, University of Missouri, Columbia, Missouri 65212, USA.
Magliola
L
L
Waters
C B
CB
Rubin
L J
LJ
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003331:Coronary Vessels / Q000201:enzymology / Q000502:physiology*; D019332:Endothelin-1 / Q000494:pharmacology*; D004789:Enzyme Activation; D005260:Female; D007473:Ion Channels / Q000502:physiology*; D007700:Kinetics; D009119:Muscle Contraction / Q000187:drug effects; D010712:Phosphatidic Acids / Q000378:metabolism; D010740:Phospholipases / Q000378:metabolism*; D013552:Swine; D010738:Type C Phospholipases / Q000494:pharmacology
D019332:Endothelin-1; D007473:Ion Channels; D010712:Phosphatidic Acids; D010740:Phospholipases; D010738:Type C Phospholipases; D002118:Calcium