Dataset Information

Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells.

ABSTRACT: TRPC5 is a member of the canonical transient receptor potential (TRPC) family of proteins that forms cationic channels either through homomultimeric assembly or heteromultimeric coordination with other TRPC proteins. It is expressed in a variety of cells including central neurones and endothelial cells and has susceptibility to stimulation by multiple factors. Here we investigated if TRPC5 is sensitive to nitric oxide. Mouse TRPC5 or human TRPC5 was over-expressed in HEK293 cells, and TRPC5 activity was determined by measuring the cytosolic Ca(2+) concentration with an indicator dye or by recording membrane current under voltage clamp. TRPC5 activity could be evoked by carbachol acting at muscarinic receptors, lanthanum, or a reducing agent. However, S-nitroso-N-acetylpenicillamine (SNAP) and diethylamine NONOate (DEA-NONOate) failed to stimulate or inhibit TRPC5 at concentrations that generated nitric oxide, caused vasorelaxation, or suppressed activity of TRPC6 via protein kinase G. At high concentrations, SNAP (but not DEA-NONOate) occasionally stimulated TRPC5 but the effect was confounded by background TRPC5-independent Ca(2+) signals. Endogenous Ca(2+)-entry in bovine aortic endothelial cells (BAECs) was suppressed by SNAP; TRPC5 blocking antibody or dominant-negative mutant TRPC5 suppressed this Ca(2+) entry and occluded the effect of SNAP. The data suggest that nitric oxide is not a direct modulator of homomeric TRPC5 channels but may inhibit endogenous BAEC channels that contain TRPC5.

SUBMITTER: Wong CO

PROVIDER: S-EPMC3622007 | biostudies-other | 2010 Jun

REPOSITORIES: biostudies-other

ACCESS DATA

Publications

Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells.

Wong Ching-On CO Sukumar Piruthivi P Beech David J DJ Yao Xiaoqiang X

Pflugers Archiv : European journal of physiology 20100601 1

TRPC5 is a member of the canonical transient receptor potential (TRPC) family of proteins that forms cationic channels either through homomultimeric assembly or heteromultimeric coordination with other TRPC proteins. It is expressed in a variety of cells including central neurones and endothelial cells and has susceptibility to stimulation by multiple factors. Here we investigated if TRPC5 is sensitive to nitric oxide. Mouse TRPC5 or human TRPC5 was over-expressed in HEK293 cells, and TRPC5 acti ...[more]

PMID: 20390293

Similar Datasets

Project description:Endothelial dysfunction in small arteries is a ubiquitous, early feature of cardiovascular disease, including hypertension. Dysfunction reflects reduced bioavailability of endothelium-derived nitric oxide (NO) and depressed endothelium-dependent hyperpolarization that enhances vasoreactivity. We measured smooth muscle membrane potential and tension, smooth muscle calcium, and used real-time quantitative polymerase chain reaction in small arteries and isolated tubes of endothelium to investigate how dysfunction enhances vasoreactivity. Rat nonmyogenic mesenteric resistance arteries developed vasomotion to micromolar phenylephrine (α1-adrenoceptor agonist); symmetrical vasoconstrictor oscillations mediated by L-type voltage-gated Ca2+ channels (VGCCs). Inhibiting NO synthesis abolished vasomotion so nanomolar phenylephrine now stimulated rapid, transient depolarizing spikes in the smooth muscle associated with chaotic vasomotion/vasospasm. Endothelium-dependent hyperpolarization block also enabled phenylephrine-vasospasm but without spikes or chaotic vasomotion. Depolarizing spikes were Ca2+-based and abolished by either T-type or L-type VGCCs blockers with depressed vasoconstriction. Removing NO also enabled transient spikes/vasoconstriction to Bay K-8644 (L-type VGCC activator). However, these were abolished by the L-type VGCC blocker nifedipine but not T-type VGCC block. Phenylephrine also initiated T-type VGCC-transient spikes and enhanced vasoconstriction after NO loss in nonmyogenic arteries from spontaneously hypertensive rats. In contrast to mesenteric arteries, myogenic coronary arteries displayed transient spikes and further vasoconstriction spontaneously on loss of NO. T-type VGCC block abolished these spikes and additional vasoconstriction but not myogenic tone. Therefore, in myogenic and nonmyogenic small arteries, reduced NO bioavailability engages T-type VGCCs, triggering transient depolarizing spikes in normally quiescent vascular smooth muscle to cause vasospasm. T-type block may offer a means to suppress vasospasm without inhibiting myogenic tone mediated by L-type VGCCs.

Project description:ATP-sensitive potassium (KATP) channels in neurons regulate excitability, neurotransmitter release and mediate protection from cell-death. Furthermore, activation of KATP channels is suppressed in DRG neurons after painful-like nerve injury. NO-dependent mechanisms modulate both KATP channels and participate in the pathophysiology and pharmacology of neuropathic pain. Therefore, we investigated NO modulation of KATP channels in control and axotomized DRG neurons.Cell-attached and cell-free recordings of KATP currents in large DRG neurons from control rats (sham surgery, SS) revealed activation of KATP channels by NO exogenously released by the NO donor SNAP, through decreased sensitivity to [ATP]i. This NO-induced KATP channel activation was not altered in ganglia from animals that demonstrated sustained hyperalgesia-type response to nociceptive stimulation following spinal nerve ligation. However, baseline opening of KATP channels and their activation induced by metabolic inhibition was suppressed by axotomy. Failure to block the NO-mediated amplification of KATP currents with specific inhibitors of sGC and PKG indicated that the classical sGC/cGMP/PKG signaling pathway was not involved in the activation by SNAP. NO-induced activation of KATP channels remained intact in cell-free patches, was reversed by DTT, a thiol-reducing agent, and prevented by NEM, a thiol-alkylating agent. Other findings indicated that the mechanisms by which NO activates KATP channels involve direct S-nitrosylation of cysteine residues in the SUR1 subunit. Specifically, current through recombinant wild-type SUR1/Kir6.2 channels expressed in COS7 cells was activated by NO, but channels formed only from truncated isoform Kir6.2 subunits without SUR1 subunits were insensitive to NO. Further, mutagenesis of SUR1 indicated that NO-induced KATP channel activation involves interaction of NO with residues in the NBD1 of the SUR1 subunit.NO activates KATP channels in large DRG neurons via direct S-nitrosylation of cysteine residues in the SUR1 subunit. The capacity of NO to activate KATP channels via this mechanism remains intact even after spinal nerve ligation, thus providing opportunities for selective pharmacological enhancement of KATP current even after decrease of this current by painful-like nerve injury.

Dataset Information

Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells.

Publications

Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets