Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells

Elisheva Rhodes-Mordov, Tal Brandwine-Shemmer, Rachel Zaguri, Rita Gutorov, Maximilian Peters, Baruch Minke*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Physiological activation by light of the Drosophila TRP and TRP-like (TRPL) channels requires the activation of phospholipase Cβ (PLC). The hydrolysis of phosphatidylinositol 4,5, bisphosphate (PIP2) by PLC is a crucial step in the still-unclear light activation, while the generation of Diacylglycerol (DAG) by PLC seems to be involved. In this study, we re-examined the ability of a DAG analogue 1-oleoyl-2-acetyl-sn-glycerol (OAG) to activate the TRPL channels expressed in HEK cells. Unlike previous studies, we added OAG into the cytosol via a patch-clamp pipette and observed robust activation of the expressed TRPL channels. However, TRPL channel activation was much slower than the physiologically activated TRPL by light. Therefore, we used a picosecond-fast optically activated DAG analogue, OptoDArG. Inactive OptoDArG was added into the intracellular solution with the patch-clamp pipette, and it slowly accumulated on the surface membrane of the recorded HEK cell in the dark. A fast application of intense UV light to the recorded cell resulted in a robust and relatively fast TRPL-dependent current that was greatly accelerated by the constitutively active TRPLF557I pore-region mutation. However, this current of the mutant channel was still considerably slower than the native light-induced TRPL current, suggesting that DAG alone is not sufficient for TRPL channel activation under physiological conditions.

Original languageEnglish
Article number6289
JournalInternational Journal of Molecular Sciences
Volume24
Issue number7
DOIs
StatePublished - Apr 2023

Bibliographical note

Publisher Copyright:
© 2023 by the authors.

Keywords

  • (OptoDArG)
  • 1-oleoyl-2-acetyl-sn-glycerol (OAG)
  • 3-hydroxypropane-1,2-diylbis(4-(4-((E)-(4-butylphenyl) diazenyl) phenyl
  • Diacylglycerol (DAG)
  • Drosophila TRP/TRPL channels
  • phospholipase C (PLC)

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