Assessment of the Role of the Inositol 1,4,5-Trisphosphate Receptor in the Activation of Transient Receptor Potential Channels and Store-operated Ca 2+ Entry Channels

Hong Tao Ma, Kartik Venkatachalam, Hong Sheng Li, Craig Montell, Tomohiro Kurosaki, Randen L. Patterson, Donald L. Gill

Research output: Contribution to journalArticle

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Abstract

The mechanism for coupling between Ca2+ stores and store-operated channels (SOCs) is an important but unresolved question. Although SOCs have not been molecularly identified, transient receptor potential (TRP) channels share a number of operational parameters with SOCs. The question of whether activation of SOCs and TRP channels is mediated by the inositol 1,4,5-trisphosphate receptor (InsP3R) was examined using the permeant InsP3R antagonist, 2-aminoethoxydiphenyl borate (2-APB) in both mammalian and invertebrate systems. In HEK293 cells stably transfected with human TRPC3 channels, the actions of 2-APB to block carbachol-induced InsP3R-mediated store release and carbachol-induced Sr2+ entry through TRPC3 channels were both reversed at high agonist levels, suggesting InsP3Rs mediate TRPC3 activation. However, electroretinogram recordings of the light-induced current in Drosophila revealed that the TRP channel-mediated responses in wild-type as well as trp and trpl mutant flies were all inhibited by 2-APB. This action of 2-APB is likely InsP3R-independent since InsP3Rs are dispensable for the light response. We used triple InsP3R knockout DT40 chicken B-cells to further assess the role of InsP3Rs in SOC activation. 45Ca2+ flux analysis revealed that although DT40 wild-type cells retained normal InsP3Rs mediating 2-APB-sensitive Ca2+ release, the DT40InsP3R-k/o cells were devoid of functional InsP3Rs. Using intact cells, all parameters of Ca 2+ store function and SOC activation were identical in DT40wt and DT40InsP3R-k/o cells. Moreover, in both cell lines SOC activation was completely blocked by 2-APB, and the kinetics of action of 2-APB on SOCs (time dependence and IC50) were identical. The results indicate that (a) the action of 2-APB on Ca2+ entry is not mediated by the InsP3R and (b) the effects of 2-APB provide evidence for an important similarity in the function of invertebrate TRP channels, mammalian TRP channels, and mammalian storeoperated channels.

Original languageEnglish (US)
Pages (from-to)18888-18896
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number22
DOIs
StatePublished - Jun 1 2001
Externally publishedYes

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Transient Receptor Potential Channels
Inositol 1,4,5-Trisphosphate Receptors
Chemical activation
Carbachol
Invertebrates
Cells
2-aminoethoxydiphenyl borate
Light
HEK293 Cells
Induced currents
Diptera
Inhibitory Concentration 50
Drosophila
Chickens
B-Lymphocytes
Fluxes
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Assessment of the Role of the Inositol 1,4,5-Trisphosphate Receptor in the Activation of Transient Receptor Potential Channels and Store-operated Ca 2+ Entry Channels. / Ma, Hong Tao; Venkatachalam, Kartik; Li, Hong Sheng; Montell, Craig; Kurosaki, Tomohiro; Patterson, Randen L.; Gill, Donald L.

In: Journal of Biological Chemistry, Vol. 276, No. 22, 01.06.2001, p. 18888-18896.

Research output: Contribution to journalArticle

Ma, Hong Tao ; Venkatachalam, Kartik ; Li, Hong Sheng ; Montell, Craig ; Kurosaki, Tomohiro ; Patterson, Randen L. ; Gill, Donald L. / Assessment of the Role of the Inositol 1,4,5-Trisphosphate Receptor in the Activation of Transient Receptor Potential Channels and Store-operated Ca 2+ Entry Channels. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 22. pp. 18888-18896.
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abstract = "The mechanism for coupling between Ca2+ stores and store-operated channels (SOCs) is an important but unresolved question. Although SOCs have not been molecularly identified, transient receptor potential (TRP) channels share a number of operational parameters with SOCs. The question of whether activation of SOCs and TRP channels is mediated by the inositol 1,4,5-trisphosphate receptor (InsP3R) was examined using the permeant InsP3R antagonist, 2-aminoethoxydiphenyl borate (2-APB) in both mammalian and invertebrate systems. In HEK293 cells stably transfected with human TRPC3 channels, the actions of 2-APB to block carbachol-induced InsP3R-mediated store release and carbachol-induced Sr2+ entry through TRPC3 channels were both reversed at high agonist levels, suggesting InsP3Rs mediate TRPC3 activation. However, electroretinogram recordings of the light-induced current in Drosophila revealed that the TRP channel-mediated responses in wild-type as well as trp and trpl mutant flies were all inhibited by 2-APB. This action of 2-APB is likely InsP3R-independent since InsP3Rs are dispensable for the light response. We used triple InsP3R knockout DT40 chicken B-cells to further assess the role of InsP3Rs in SOC activation. 45Ca2+ flux analysis revealed that although DT40 wild-type cells retained normal InsP3Rs mediating 2-APB-sensitive Ca2+ release, the DT40InsP3R-k/o cells were devoid of functional InsP3Rs. Using intact cells, all parameters of Ca 2+ store function and SOC activation were identical in DT40wt and DT40InsP3R-k/o cells. Moreover, in both cell lines SOC activation was completely blocked by 2-APB, and the kinetics of action of 2-APB on SOCs (time dependence and IC50) were identical. The results indicate that (a) the action of 2-APB on Ca2+ entry is not mediated by the InsP3R and (b) the effects of 2-APB provide evidence for an important similarity in the function of invertebrate TRP channels, mammalian TRP channels, and mammalian storeoperated channels.",
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