Complex regulation of the TRPC3, 6 and 7 channel subfamily by diacylglycerol and phosphatidylinositol-4,5-bisphosphate

doi:10.1016/j.ceca.2007.09.001

Cell Calcium

Volume 43, Issue 5, May 2008, Pages 506-514

https://doi.org/10.1016/j.ceca.2007.09.001 Get rights and content

Abstract

TRPC3, 6 and 7 channels constitute a subgroup of non-selective, calcium-permeable cation channels within the TRP superfamily that are activated by products of phospholipase C-mediated breakdown of phosphatidylinositol-4,5-bisphosphate (PIP₂). A number of ion channels, including other members of the TRP superfamily, are regulated directly by PIP₂. However, there is little information on the regulation of the TRPC channel subfamily by PIP₂. Pretreatment of TRPC7-expressing cells with a drug that blocks the synthesis of polyphosphoinositides inhibited the ability of the synthetic diacylglycerol, oleyl-acetyl glycerol, to activate TRPC7. In excised patches, TRPC7 channels were robustly activated by application of PIP₂ or ATP, but not by inositol 1,4,5-trisphosphate. Similar results were obtained with TRPC6 and TRPC3, although the effects of PIP₂ were somewhat less and with TRPC3 there was no significant effect of ATP. In the cell-attached configuration, TRPC7 channels could be activated by the synthetic diacylglycerol analog, oleyl-acetyl glycerol. However, this lipid mediator did not activate TRPC7 channels in excised patches. In addition, channel activation by PIP₂ in excised patches was significantly greater than that observed with oleyl-acetyl glycerol in the cell-attached configuration. These findings reveal complex regulation of TRPC channels by lipid mediators. The results also reveal for the first time direct activation by PIP₂ of members of the TRPC ion channel subfamily.

Section snippets

Methods

Experiments were carried out with HEK293 cells stably expressing either TRPC3, TRPC6 or TRPC7 [9], [10], [11]. Ca²⁺ measurements were carried out with TRPC7-expressing cells loaded with Fura-2-AM and a cell imaging system (Intracellular Imaging, Inc., Cincinnati, OH) as described previously [12]. For Fura-2 measurements, the ratio obtained by dividing the emitted fluorescence from excitation at 340 nm by the emitted fluorescence measured after excitation at 380 nm is reported as an indicator of

Results

As previously demonstrated [15], [16], in HEK293 cells stably expressing TRPC7, application of the membrane permeant diacylglycerol analog, oleyl-acetyl glycerol (OAG) activated Ca²⁺ entry (Fig. 1A). Pretreatment of the cells with the phosphoinositide lipid kinase inhibitor, LY294002 [17], [18] caused near total inhibition of the [Ca²⁺]_i signal (Fig. 1A). This inhibition does not likely result from a direct action on TRPC7 channels, because addition of LY294002 after the Ca²⁺ signal had

Discussion

A wide variety of ion channels and plasma membrane transporters are known to be regulated by the acidic phospholipid, PIP₂ [7], [8]. Generally, this modulation is a positive one. Since PIP₂ is constitutively present in the membrane of cells and can be decreased by activation of PLC-coupled receptors, this may provide a means for negative regulation of ion channel signaling pathways. That appears to be the case for a number of channels in the TRP superfamily that are activated by PIP₂ and

Acknowledgements

Drs. David Armstrong, Jerrel Yakel and Guillermo Vazquez read the manuscript and provided helpful comments. This work was supported by funds from the Intramural Program of the NIEHS and NIH.

References (37)

M. Trebak et al.
The TRPC3/6/7 subfamily of cation channels
Cell Calcium
(2003)
M. Trebak et al.
Signaling mechanism for receptor-activated TRPC3 channels
J. Biol. Chem.
(2003)
B.C. Suh et al.
Regulation of ion channels by phosphatidylinositol-4,5-bisphosphate
Curr. Opin. Neurobiol.
(2005)
C. Patton et al.
Some precautions in using chelators to buffer metals in biological solutions
Cell Calcium
(2004)
G. Vazquez et al.
Native TRPC7 channel activation by an inositol trisphosphate receptor-dependent mechanism
J. Biol. Chem.
(2006)
T. Okada et al.
Molecular and functional characterization of a novel mouse transient receptor potential protein homologue TRP7. Ca²⁺-permeable cation channel that is constitutively activated and enhanced by stimulation of G protein-coupled receptor
J. Biol. Chem.
(1999)
S. Jung et al.
Lanthanides potentiate TRPC5 currents by an action at extracellular sites close to the pore mouth
J. Biol. Chem.
(2003)
R.C. Hardie et al.
Calcium influx via TRP channels is reguired to maintain PIP2 levels in Drosophila photoreceptors
Neuron
(2001)
G. Vazquez et al.
The mammalian TRPC cation channels
Biochim. Biophys. Acta
(2004)
Y. Kwon et al.
Integration of phosphoinositide- and calmodulin-mediated regulation of TRPC6
Mol. Cell
(2007)

B.Z. Peterson et al.

Calmodulin is the Ca²⁺ sensor for Ca²⁺-dependent inactivation of L-type calcium channels

Neuron

(1999)

G. Vazquez et al.

Obligatory role of Src kinase in the signaling mechanism for TRPC3 cation channels

J. Biol. Chem.

(2004)

D.E. Clapham et al.

The TRP ion channel family

Nat. Rev. Neurosci.

(2002)

R.C. Hardie

Regulation of trp channels via lipid second messengers

Ann. Rev. Physiol.

(2003)

A. Dietrich et al.

Functional characterization and physiological relevance of the TRPC3/6/7 subfamily of cation channels

Naunyn Schmiedebergs Arch. Pharmacol.

(2005)

K. Groschner et al.

TRPC3: a versatile transducer molecule that serves integration and diversification of cellular signals

Naunyn Schmiedebergs Arch. Pharmacol.

(2005)

D.W. Hilgemann et al.

The complex and intriguing lives of PIP2 with ion channels and transporters

Science's STKE

(2001)

R.R. McKay et al.

Cloning and expression of the human transient receptor potential 4 (TRP4) gene: localization and functional expression of human TRP4 and TRP3

Biochem. J.

(2000)

Cited by (109)

Structural mechanism of human TRPC3 and TRPC6 channel regulation by their intracellular calcium-binding sites
2022, Neuron
TRPC3 and TRPC6 channels are calcium-permeable non-selective cation channels that are involved in many physiological processes. The gain-of-function (GOF) mutations of TRPC6 lead to familial focal segmental glomerulosclerosis (FSGS) in humans, but their pathogenic mechanism remains elusive. Here, we report the cryo-EM structures of human TRPC3 in both high-calcium and low-calcium conditions. Based on these structures and accompanying electrophysiological studies, we identified both inhibitory and activating calcium-binding sites in TRPC3 that couple intracellular calcium concentrations to the basal channel activity. These calcium sensors are also structurally and functionally conserved in TRPC6. We uncovered that the GOF mutations of TRPC6 activate the channel by allosterically abolishing the inhibitory effects of intracellular calcium. Furthermore, structures of human TRPC6 in complex with two chemically distinct inhibitors bound at different ligand-binding pockets reveal different conformations of the transmembrane domain, providing templates for further structure-based drug design targeting TRPC6-related diseases such as FSGS.
Intrinsic properties of primary hippocampal neurons contribute to PIP<inf>2</inf> depletion during nsEP-induced physiological response
2021, Bioelectrochemistry
High-energy, short-duration electric pulses (EPs) are known to be effective in neuromodulation, but the biological mechanisms underlying this effect remain unclear. Recently, we discovered that nanosecond electric pulses (nsEPs) could initiate the phosphatidylinositol_4,5-bisphosphate (PIP₂) depletion in non-excitable cells identical to agonist-induced activation of the G_q11 coupled receptors. PIP₂ is the precursor for multiple intracellular second messengers critically involved in the regulation of intracellular Ca²⁺ homeostasis and plasma membrane (PM) ion channels responsible for the control of neuronal excitability. In this paper we demonstrate a novel finding that five day in vitro (DIV5) primary hippocampal neurons (PHNs) undergo significantly higher PIP₂ depletion after 7.5 kV/cm 600 ns EP exposure than DIV1 PHNs and day 1–5 (D1-D5) non-excitable Chinese hamster ovarian cells with muscarinic receptor 1 (CHO-hM₁). Despite the age of development, the stronger 15 kV/cm 600 ns or longer 7.5 kV/cm 12 µs EP initiated profound PIP₂ depletion in all cells studied, outlining damage of the cellular PM and electroporation. Therefore, the intrinsic properties of PHNs in concert with nanoporation explain the stronger neuronal response to nsEP at lower intensity exposures. PIP₂ reduction in neurons could be a primary biological mechanism responsible for the stimulation or inhibition of neuronal tissues.
Phosphoinositides and calcium signaling; a marriage arranged at ER-PM contact sites
2020, Current Opinion in Physiology
Calcium (Ca²⁺) ions are critically important in orchestrating countless regulatory processes in eukaryotic cells. Consequently, cells tightly control cytoplasmic Ca²⁺ concentrations using a complex array of Ca²⁺-selective ion channels, transporters, and signaling effectors. Ca²⁺ transport across various cellular membranes is highly dependent on the intrinsic properties of specific membrane compartments and conversely, local Ca²⁺ changes have profound effects on the membrane lipid composition of such membrane sub-domains. In particular, inositol phospholipids are a minor class of phospholipids that play pivotal roles in the control of Ca²⁺-dependent signaling pathways. In this review, we will highlight some of the recent advances in this field as well as their impact in defining future research directions.
The involvement of TRP channels in memory formation and task retrieval in a passive avoidance task in one-day old chicks
2020, Neurobiology of Learning and Memory
An increase in the intracellular Ca2+ level in neurons is one of the main steps in the memory formation cascade. The increase results from extracellular Ca2+ influx by activation of ionotropic glutamate receptors and release from intracellular stores by the stimulation of IP3 receptors (IP3Rs) via group I metabotropic glutamate receptors (mGluR1/5). Recent data indicate an additional mechanism resulting in Ca2+ influx into neurons, triggered by intracellular signals that are directly connected to the activation of group I mGluRs. This influx occurs through transient receptor potential (TRP) channels, which are permeable to Na+, K+ and, mainly, Ca2+. These channels are activated by increases in intracellular Ca2+, diacylglycerol (DAC) and inositol 1,4,5-triphosphate (IP3) level resulting from a group I mGluR activation. The aim of the present study was to investigate the participation of TRP channels, especially from TRPC and TRPV groups, in memory consolidation and reconsolidation and memory retrieval processes in a passive avoidance task in one-day old chicks. TRP channels were blocked by the injection of the unspecific channel modulators SKF 96365 (2.5 µl 30 µM/hemisphere) and 2-APB (2.5 µl 10 µM/hemisphere) directly into the intermediate medial mesopallium (IMM) region of the chick brain immediately after initial training or after a reminder. The inhibition of specific TRP channels (TRPV1, TRPV3 or TRPC3) was achieved by the application of selective antibodies. Our results demonstrate that the inhibition of TRP channels by the application of both modulators disrupted memory consolidation, resulting in permanent task amnesia. The inhibition of the TRPV1, TRPC3 and TRPV3 channels by specific antibodies resulted in similar amnesia. Moreover, the inhibition of TRP channels by SKF 96365 and 2-APB at different time points after initial training or after the reminder also resulted in amnesia, indicating the role of TRP channels in memory retrieval.
The inhibition of calcium influx through these channels resulted in permanent memory disruption, which suggests that the calcium signal generated by TRP channels is crucial for memory formation and retrieval processes. For the first time, the important role of TRPV3 channels in memory formation was demonstrated.
TRPC channels: Structure, function, regulation and recent advances in small molecular probes
2020, Pharmacology and Therapeutics
Citation Excerpt :
For other TRPC channels, PIP2 dependence has been reported for native channels in vascular smooth muscle cells likely composed of TRPC1/C5 heteromers, where TRPC1 appeared to confer the PIP2 regulation (Shi et al., 2012). In heterologous systems, PIP2 activated overexpressed TRPC6 and C7 in inside-out patches (Lemonnier, Trebak, & Putney Jr, 2008), but depleting PIP2 in rabbit mesenteric artery smooth muscle cells augmented endogenous TRPC6-like activity (Albert, Saleh, & Large, 2008). With the use of membrane depolarization in whole-cell recordings to dephosphorylate PIP2 through voltage-sensitive phosphatases, PIP2 was found to be essential in supporting TRPC3, C6, and C7 function, and the three TRPC members exhibited differences in the sensitivity to PIP2 (Imai, Itsuki, Okamura, Inoue, & Mori, 2012).
Transient receptor potential canonical (TRPC) channels constitute a group of receptor-operated calcium-permeable nonselective cation channels of the TRP superfamily. The seven mammalian TRPC members, which can be further divided into four subgroups (TRPC1, TRPC2, TRPC4/5, and TRPC3/6/7) based on their amino acid sequences and functional similarities, contribute to a broad spectrum of cellular functions and physiological roles. Studies have revealed complexity of their regulation involving several components of the phospholipase C pathway, G_i and G_o proteins, and internal Ca²⁺ stores. Recent advances in cryogenic electron microscopy have provided several high-resolution structures of TRPC channels. Growing evidence demonstrates the involvement of TRPC channels in diseases, particularly the link between genetic mutations of TRPC6 and familial focal segmental glomerulosclerosis. Because TRPCs were discovered by the molecular identity first, their pharmacology had lagged behind. This is rapidly changing in recent years owning to great efforts from both academia and industry. A number of potent tool compounds from both synthetic and natural products that selective target different subtypes of TRPC channels have been discovered, including some preclinical drug candidates. This review will cover recent advancements in the understanding of TRPC channel regulation, structure, and discovery of novel TRPC small molecular probes over the past few years, with the goal of facilitating drug discovery for the study of TRPCs and therapeutic development.
Monotherapy of RAAS blockers and mobilization of aldosterone: A mechanistic perspective study in kidney disease
2020, Chemico-Biological Interactions
In patients with acute kidney injury progressively converting into chronic kidney disease (CKD), proteinuria and high blood pressure predict progression to end-stage renal disease (ESRD). Although, Renin-angiotensin-aldosterone system (RAAS) regulates blood pressure and kidney disease through both direct and indirect mechanisms. RAAS blockers that act at the level of angiotensin or lower in the cascade can cause compensatory increases in the plasma renin and angiotensin II level. Here, in this review article, we are exploring the evidence-based on RAAS blockade action releases of aldosterone and hypothesizing the molecular mechanism for converting the acute kidney injury into chronic kidney disease to end-stage renal disease.

View all citing articles on Scopus

¹: Present address: Inserm U800, Universite de Lille 1, Villeneuve d’Ascq F-59650, France.

²: Present address: Center for Cardiovascular Sciences, Albany Medical Center, 47 New Scotland Avenue, Albany, New York 12208, United States.

³: These authors contributed equally to this work.

View full text

Published by Elsevier Ltd.

Complex regulation of the TRPC3, 6 and 7 channel subfamily by diacylglycerol and phosphatidylinositol-4,5-bisphosphate

Abstract

Section snippets

Methods

Results

Discussion

Acknowledgements

Cell Calcium

J. Biol. Chem.

Curr. Opin. Neurobiol.

Cell Calcium

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Neuron

Biochim. Biophys. Acta

Mol. Cell

Neuron

J. Biol. Chem.

The TRP ion channel family

Nat. Rev. Neurosci.

Regulation of trp channels via lipid second messengers

Ann. Rev. Physiol.

Functional characterization and physiological relevance of the TRPC3/6/7 subfamily of cation channels

Naunyn Schmiedebergs Arch. Pharmacol.

TRPC3: a versatile transducer molecule that serves integration and diversification of cellular signals

Naunyn Schmiedebergs Arch. Pharmacol.

The complex and intriguing lives of PIP2 with ion channels and transporters

Science's STKE

Cloning and expression of the human transient receptor potential 4 (TRP4) gene: localization and functional expression of human TRP4 and TRP3

Biochem. J.