Contraction of rat thoracic aorta strips by endothelin-1 in the absence of extracellular Ca2+.
1. Endothelin-1 (ET-1) caused a concentration-dependent contraction of helical strips from rat thoracic aorta in the absence of extracellular Ca2+. The Ca(2+)-depleted muscle strips, prepared by three repeated applications of 10(-2) M caffeine or 10(-6) M noradrenaline in Ca(2+)-free buffer, were contracted by 10(-8) M ET-1 in the same manner as non-treated strips. 2. In the absence of extracellular Ca2+, 10(-7) M phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, induced a small but sustained contraction of the rat thoracic aorta strips within 60 min. Preincubation of the strips with 10(-7) M PMA for 60 min in Ca(2+)-free buffer, did not affect the 10(-8) M ET-1-induced contraction, but decreased the 5 x 10(-8) M phorbol 12,13-dibutyrate (PDB)-, or the 10(-7) M PMA-induced contraction, and potentiated the contraction induced by 10(-8) M urotensin II. Preincubation with 10(-8) M ET-1 (which induced maximum contraction) for 25 min in Ca(2+)-free buffer did not change the subsequent contraction induced by PMA (10(-7) M) or urotensin II (10(-8) M) but gave a somewhat lower maximum tension than in non-treated strips. 3. Calyculin-A, a potent inhibitor of phosphatase, also induced a contraction of the Ca(2+)-depleted muscle strips in Ca(2+)-free buffer. Preincubation of the strips with ET-1 (10(-8) M) or PMA (10(-7) M) decreased the calyculin-A (3 x 10(-8) M)-induced contraction.4. These results suggest that ET-1 may induce phosphorylation of an unknown protein either without an increase in myoplasmic Ca2 + concentration or, alternatively, with mobilization of intracellular Ca2+ from noradrenaline- and caffeine-insensitive Ca2 + sources, through a mechanism different from that of phorbol ester.
Itoh
H
H
First Department of Internal Medicine, Faculty of Medicine, University of Mie, Japan.
Higuchi
H
H
Hiraoka
N
N
Ito
M
M
Konishi
T
T
Nakano
T
T
Lederis
K
K
D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000502:physiology*; D016232:Endothelins / Q000494:pharmacology*; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008297:Male; D008387:Marine Toxins; D009119:Muscle Contraction / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D009638:Norepinephrine / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014579:Urotensins / Q000494:pharmacology; D014662:Vasoconstrictor Agents / Q000494:pharmacology
D016232:Endothelins; D007202:Indicators and Reagents; D008387:Marine Toxins; D010080:Oxazoles; D014579:Urotensins; D014662:Vasoconstrictor Agents; D015240:Phorbol 12,13-Dibutyrate; D002110:Caffeine; C059041:calyculin A; C029677:urotensin II; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium; D009638:Norepinephrine