P2X receptor-stimulated calcium responses in preglomerular vascular smooth muscle cells involves 20-hydroxyeicosatetraenoic acid

Xueying Zhao, J R Falck, V. Raj Gopal, Edward W. Inscho, John D. Imig

Research output: Contribution to journalArticle

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Abstract

The current study tested the hypothesis that endogenous 20-hydroxyeicosatetraenoic acid (20-HETE) contributes to the increase in intracellular calcium ([Ca2+]i) elicited by P2X receptor activation in renal microvascular smooth muscle cells. Vascular smooth muscle cells obtained from rats were loaded with fura-2 and studied using standard single cell fluorescence microscopy. Basal renal myocyte [Ca2+] i averaged 96 ± 5 nM. ATP (10 and 100 μM) increased vascular smooth muscle cell [Ca2+]i by 340 ± 88 and 555 ± 80 nM, respectively. The cytochrome P450 hydroxylase inhibitor, N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS), or the 20-HETE antagonist, 20-hydroxyeicosa-6(Z), 15(Z)-dienoic acid (20-HEDE), significantly attenuated the peak myocyte [Ca2+]i responses to 10 and 100 μM ATP. ATP (100 μM) increased vascular smooth muscle cell [Ca 2+]i by 372 ± 93 and 163 ± 55 nM in the presence of DDMS or 20-HEDE, respectively. The P2X receptor agonist, α,β-methyleneylene-ATP (10 μM), increased myocyte [Ca 2+]i by 78 ± 12 nM, and this response was significantly attenuated by DDMS (40 ± 15 nM). In contrast, the vascular smooth muscle cell [Ca2+]i evoked by the P2Y agonist, UTP (100 μM), was not altered by DDMS or 20-HEDE. The effect of 20-HETE on [Ca2+]i was also assessed, and the peak increases in [Ca2+]i averaged 62 ± 12 and 146 ± 70 nM at 20-HETE concentrations of 1 and 10 μM, respectively. These results demonstrate that 20-HETE plays a significant role in the renal microvascular smooth muscle cell [Ca2+]i response to P2X receptor activation.

Original languageEnglish (US)
Pages (from-to)1211-1217
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume311
Issue number3
DOIs
StatePublished - Dec 2004

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Calcium-Sensing Receptors
Vascular Smooth Muscle
Smooth Muscle Myocytes
Adenosine Triphosphate
Muscle Cells
Kidney
Uridine Triphosphate
Fura-2
Mixed Function Oxygenases
Fluorescence Microscopy
Cytochrome P-450 Enzyme System
20-hydroxy-5,8,11,14-eicosatetraenoic acid
DDMS
Calcium
20-hydroxyeicosa-6(Z),15(Z)-dienoic acid

ASJC Scopus subject areas

  • Pharmacology

Cite this

P2X receptor-stimulated calcium responses in preglomerular vascular smooth muscle cells involves 20-hydroxyeicosatetraenoic acid. / Zhao, Xueying; Falck, J R; Gopal, V. Raj; Inscho, Edward W.; Imig, John D.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 311, No. 3, 12.2004, p. 1211-1217.

Research output: Contribution to journalArticle

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abstract = "The current study tested the hypothesis that endogenous 20-hydroxyeicosatetraenoic acid (20-HETE) contributes to the increase in intracellular calcium ([Ca2+]i) elicited by P2X receptor activation in renal microvascular smooth muscle cells. Vascular smooth muscle cells obtained from rats were loaded with fura-2 and studied using standard single cell fluorescence microscopy. Basal renal myocyte [Ca2+] i averaged 96 ± 5 nM. ATP (10 and 100 μM) increased vascular smooth muscle cell [Ca2+]i by 340 ± 88 and 555 ± 80 nM, respectively. The cytochrome P450 hydroxylase inhibitor, N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS), or the 20-HETE antagonist, 20-hydroxyeicosa-6(Z), 15(Z)-dienoic acid (20-HEDE), significantly attenuated the peak myocyte [Ca2+]i responses to 10 and 100 μM ATP. ATP (100 μM) increased vascular smooth muscle cell [Ca 2+]i by 372 ± 93 and 163 ± 55 nM in the presence of DDMS or 20-HEDE, respectively. The P2X receptor agonist, α,β-methyleneylene-ATP (10 μM), increased myocyte [Ca 2+]i by 78 ± 12 nM, and this response was significantly attenuated by DDMS (40 ± 15 nM). In contrast, the vascular smooth muscle cell [Ca2+]i evoked by the P2Y agonist, UTP (100 μM), was not altered by DDMS or 20-HEDE. The effect of 20-HETE on [Ca2+]i was also assessed, and the peak increases in [Ca2+]i averaged 62 ± 12 and 146 ± 70 nM at 20-HETE concentrations of 1 and 10 μM, respectively. These results demonstrate that 20-HETE plays a significant role in the renal microvascular smooth muscle cell [Ca2+]i response to P2X receptor activation.",
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