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

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 (PIP(2)) by PLC is a crucial step in the still-unclear light activation, while the generation of Diacylglyc...

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Autores principales: Rhodes-Mordov, Elisheva, Brandwine-Shemmer, Tal, Zaguri, Rachel, Gutorov, Rita, Peters, Maximilian, Minke, Baruch
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10093889/
https://www.ncbi.nlm.nih.gov/pubmed/37047261
http://dx.doi.org/10.3390/ijms24076289
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author Rhodes-Mordov, Elisheva
Brandwine-Shemmer, Tal
Zaguri, Rachel
Gutorov, Rita
Peters, Maximilian
Minke, Baruch
author_facet Rhodes-Mordov, Elisheva
Brandwine-Shemmer, Tal
Zaguri, Rachel
Gutorov, Rita
Peters, Maximilian
Minke, Baruch
author_sort Rhodes-Mordov, Elisheva
collection PubMed
description 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 (PIP(2)) 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 TRPL(F557I) 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.
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spelling pubmed-100938892023-04-13 Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells Rhodes-Mordov, Elisheva Brandwine-Shemmer, Tal Zaguri, Rachel Gutorov, Rita Peters, Maximilian Minke, Baruch Int J Mol Sci Article 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 (PIP(2)) 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 TRPL(F557I) 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. MDPI 2023-03-27 /pmc/articles/PMC10093889/ /pubmed/37047261 http://dx.doi.org/10.3390/ijms24076289 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rhodes-Mordov, Elisheva
Brandwine-Shemmer, Tal
Zaguri, Rachel
Gutorov, Rita
Peters, Maximilian
Minke, Baruch
Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells
title Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells
title_full Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells
title_fullStr Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells
title_full_unstemmed Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells
title_short Diacylglycerol Activates the Drosophila Light Sensitive Channel TRPL Expressed in HEK Cells
title_sort diacylglycerol activates the drosophila light sensitive channel trpl expressed in hek cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10093889/
https://www.ncbi.nlm.nih.gov/pubmed/37047261
http://dx.doi.org/10.3390/ijms24076289
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