Expression of TRPC3 in Chinese hamster ovary cells results in calcium- activated cation currents not related to store depletion

Christof Zitt, Alexander Obukhov, Carsten Strübing, Andrea Zobel, Frank Kalkbrenner, Andreas Lückhoff, Günter Schultz

Research output: Contribution to journalArticle

213 Citations (Scopus)

Abstract

TRPC3 (or Htrp3) is a human member of the trp family of Ca2+-permeable cation channels. Since expression of TRPC3 cDNA results in markedly enhanced Ca2+ influx in response to stimulation of membrane receptors linked to phospholipase C (Zhu, X., J. Meisheng, M. Peyton, G. Bouley, R. Hurst, E. Stefani, and L. Birnbaumer. 1996. Cell. 85:661-671), we tested whether TRPC3 might represent a Ca2+ entry pathway activated as a consequence of depletion of intracellular calcium stores. CHO cells expressing TRPC3 after intranuclear injection of cDNA coding for TRPC3 were identified by fluorescence from green fluorescent protein. Expression of TRPC3 produced cation currents with little selectivity for Ca2+ over Na+. These currents were constitutively active, not enhanced by depletion of calcium stores with inositol-1,4,5-trisphosphate or thapsigargin, and attenuated by strong intracellular Ca2+ buffering. Ionomycin led to profound increases of currents, but this effect was strictly dependent on the presence of extracellular Ca2+. Likewise, infusion of Ca2+ into cell through the patch pipette increased TRPC3 currents. Therefore, TRPC3 is stimulated by a Ca2+-dependent mechanism. Studies on TRPC3 in inside-out patches showed cation-selective channels with 60-pS conductance and short (2+ concentration on the cytosolic side of inside-out patches (from 0 to 1 and 30 μM), however, failed to stimulate channel activity, even in the presence of calmodulin (0.2 μM). We conclude that TRPC3 codes for a Ca2+permeable channel that supports Ca2+-induced Ca2+entry but should not be considered store operated.

Original languageEnglish (US)
Pages (from-to)1333-1341
Number of pages9
JournalJournal of Cell Biology
Volume138
Issue number6
DOIs
StatePublished - Sep 22 1997
Externally publishedYes

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Cricetulus
Cations
Ovary
Calcium
Complementary DNA
Ionomycin
Inositol 1,4,5-Trisphosphate
Thapsigargin
CHO Cells
Type C Phospholipases
Calmodulin
Green Fluorescent Proteins
Fluorescence
Injections
Membranes

ASJC Scopus subject areas

  • Cell Biology

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Expression of TRPC3 in Chinese hamster ovary cells results in calcium- activated cation currents not related to store depletion. / Zitt, Christof; Obukhov, Alexander; Strübing, Carsten; Zobel, Andrea; Kalkbrenner, Frank; Lückhoff, Andreas; Schultz, Günter.

In: Journal of Cell Biology, Vol. 138, No. 6, 22.09.1997, p. 1333-1341.

Research output: Contribution to journalArticle

Zitt, Christof ; Obukhov, Alexander ; Strübing, Carsten ; Zobel, Andrea ; Kalkbrenner, Frank ; Lückhoff, Andreas ; Schultz, Günter. / Expression of TRPC3 in Chinese hamster ovary cells results in calcium- activated cation currents not related to store depletion. In: Journal of Cell Biology. 1997 ; Vol. 138, No. 6. pp. 1333-1341.
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abstract = "TRPC3 (or Htrp3) is a human member of the trp family of Ca2+-permeable cation channels. Since expression of TRPC3 cDNA results in markedly enhanced Ca2+ influx in response to stimulation of membrane receptors linked to phospholipase C (Zhu, X., J. Meisheng, M. Peyton, G. Bouley, R. Hurst, E. Stefani, and L. Birnbaumer. 1996. Cell. 85:661-671), we tested whether TRPC3 might represent a Ca2+ entry pathway activated as a consequence of depletion of intracellular calcium stores. CHO cells expressing TRPC3 after intranuclear injection of cDNA coding for TRPC3 were identified by fluorescence from green fluorescent protein. Expression of TRPC3 produced cation currents with little selectivity for Ca2+ over Na+. These currents were constitutively active, not enhanced by depletion of calcium stores with inositol-1,4,5-trisphosphate or thapsigargin, and attenuated by strong intracellular Ca2+ buffering. Ionomycin led to profound increases of currents, but this effect was strictly dependent on the presence of extracellular Ca2+. Likewise, infusion of Ca2+ into cell through the patch pipette increased TRPC3 currents. Therefore, TRPC3 is stimulated by a Ca2+-dependent mechanism. Studies on TRPC3 in inside-out patches showed cation-selective channels with 60-pS conductance and short (2+ concentration on the cytosolic side of inside-out patches (from 0 to 1 and 30 μM), however, failed to stimulate channel activity, even in the presence of calmodulin (0.2 μM). We conclude that TRPC3 codes for a Ca2+permeable channel that supports Ca2+-induced Ca2+entry but should not be considered store operated.",
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AU - Zitt, Christof

AU - Obukhov, Alexander

AU - Strübing, Carsten

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AU - Kalkbrenner, Frank

AU - Lückhoff, Andreas

AU - Schultz, Günter

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AB - TRPC3 (or Htrp3) is a human member of the trp family of Ca2+-permeable cation channels. Since expression of TRPC3 cDNA results in markedly enhanced Ca2+ influx in response to stimulation of membrane receptors linked to phospholipase C (Zhu, X., J. Meisheng, M. Peyton, G. Bouley, R. Hurst, E. Stefani, and L. Birnbaumer. 1996. Cell. 85:661-671), we tested whether TRPC3 might represent a Ca2+ entry pathway activated as a consequence of depletion of intracellular calcium stores. CHO cells expressing TRPC3 after intranuclear injection of cDNA coding for TRPC3 were identified by fluorescence from green fluorescent protein. Expression of TRPC3 produced cation currents with little selectivity for Ca2+ over Na+. These currents were constitutively active, not enhanced by depletion of calcium stores with inositol-1,4,5-trisphosphate or thapsigargin, and attenuated by strong intracellular Ca2+ buffering. Ionomycin led to profound increases of currents, but this effect was strictly dependent on the presence of extracellular Ca2+. Likewise, infusion of Ca2+ into cell through the patch pipette increased TRPC3 currents. Therefore, TRPC3 is stimulated by a Ca2+-dependent mechanism. Studies on TRPC3 in inside-out patches showed cation-selective channels with 60-pS conductance and short (2+ concentration on the cytosolic side of inside-out patches (from 0 to 1 and 30 μM), however, failed to stimulate channel activity, even in the presence of calmodulin (0.2 μM). We conclude that TRPC3 codes for a Ca2+permeable channel that supports Ca2+-induced Ca2+entry but should not be considered store operated.

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