GVI phospholipase A2 role in the stimulatory effect of sphingosine-1-phosphate on TRPC5 cationic channels

Eman AL-Shawaf, Sarka Tumova, Jacqueline Naylor, Yasser Majeed, Jing Li, David J. Beech

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The Transient Receptor Potential Canonical 5 (TRPC5) protein forms calcium-permeable cationic channels that are stimulated by G protein-coupled receptor agonists. The signaling pathways of such agonist effects are poorly understood. Here we investigated the potential for involvement of lysophosphatidylcholine (LPC) and arachidonic acid generated by group 6 (GVI) phospholipase A2 (PLA2) enzymes, focusing on stimulation of TRPC5 by sphingosine-1-phosphate (S1P) which acts via a pertussis toxin-sensitive (Gi/o protein) pathway without Ca2+-release. Experiments were on HEK 293 cells containing conditional expression of human TRPC5. Channel activity was recorded using an intracellular calcium indicator or whole-cell patch-clamp and PLA2 activity was detected using 3H-arachidonic acid. S1P stimulated PLA2 and TRPC5 activities. Both effects were suppressed by the GVI PLA2 inhibitor bromoenol lactone. Knock-down of GVI PLA2 by RNA interference suppressed channel activity evoked by S1P whereas activity evoked by the direct channel stimulator LPC was unaffected. Arachidonic acid did not stimulate the channels. Prior exposure of channels to LPC but not arachidonic acid suppressed channel activity evoked by S1P but not gadolinium, a putative direct stimulator of the channels. The data suggest roles of LPC and GVI PLA2 in S1P-evoked TRPC5 activity.

Original languageEnglish
Pages (from-to)343-350
Number of pages8
JournalCell Calcium
Volume50
Issue number4
DOIs
Publication statusPublished - 1 Jan 2011
Externally publishedYes

Fingerprint

Phospholipases A2
Lysophosphatidylcholines
Arachidonic Acid
Phospholipase A2 Receptors
Calcium
HEK293 Cells
Pertussis Toxin
Gadolinium
G-Protein-Coupled Receptors
RNA Interference
Proteins
sphingosine 1-phosphate
Enzymes

Keywords

  • Calcium-permeable channel
  • Cationic channel
  • Lysophosphatidylcholine
  • Phospholipase
  • Sphingosine-1-phosphate
  • Transient receptor potential

ASJC Scopus subject areas

  • Physiology
  • Molecular Biology
  • Cell Biology

Cite this

GVI phospholipase A2 role in the stimulatory effect of sphingosine-1-phosphate on TRPC5 cationic channels. / AL-Shawaf, Eman; Tumova, Sarka; Naylor, Jacqueline; Majeed, Yasser; Li, Jing; Beech, David J.

In: Cell Calcium, Vol. 50, No. 4, 01.01.2011, p. 343-350.

Research output: Contribution to journalArticle

AL-Shawaf, Eman ; Tumova, Sarka ; Naylor, Jacqueline ; Majeed, Yasser ; Li, Jing ; Beech, David J. / GVI phospholipase A2 role in the stimulatory effect of sphingosine-1-phosphate on TRPC5 cationic channels. In: Cell Calcium. 2011 ; Vol. 50, No. 4. pp. 343-350.
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