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Role of Transient Receptor Potential Channel 1 (TRPC1) in Glutamate-Induced Cell Death in the Hippocampal Cell Line HT22

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Abstract

Transient receptor potential channel 1 (TRPC1; a cation channel activated by store depletion and/or through an intracellular messenger) is expressed in a variety of tissues, including the brain. To study the physiological function of TRPC1, we investigated the role of endogenously expressed TRPC1 in glutamate-induced cell death, using the murine hippocampal cell line HT22. Knocking down TRPC1 mRNA using TRPC1-shRNA or blocking of TRPC channels using 2-APB (≥200 μM) robustly attenuated glutamate-induced cell death after 24 h of incubation with 5 mM glutamate. Glutamate toxicity in HT22 cells seems to involve metabotropic glutamate receptor mGluR5 since MPEP (2-methyl-6-(phenylethynyl)-pyridine), an mGluR5 antagonist (≥100 μM), abrogated glutamate toxicity. Furthermore, a direct activation of mGluR5 by CHPG [(RS)-chloro-5-hydroxyphenylglycine; 100 μM or 300 μM] promoted HT22 cell death. TRPC1 knock-down markedly reduced CHPG-induced cell death. These observations suggest that glutamate-induced cell death in HT22 cells activates mGluR5 receptors, which significantly increases Ca2+ influx through TRPC1 channels. TRPC1 knock-down prevented glutamate- and CHPG-induced cell death, suggesting that glutamate-induced toxicity in HT22 cells is mediated through TRPC1 channels and an mGluR5-dependent pathway. Together, this work provides evidence for a novel receptor activation pathway of TRPC1 in glutamate-induced toxicity.

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Abbreviations

2-APB:

2-Aminoethoxydiphenyl borate

CHPG:

(RS)-Chloro-5-hydroxyphenylglycine

DAG:

Diacylglycerol

glu:

Glutamate

IP3 :

Inositol triphosphate

mGluR:

Metabotropic receptors

MPEP:

2-Methyl-6-(phenylethynyl)-pyridine

PIP2 :

Phosphatidylinositol biphosphate

PLC:

Phospholipase C

ROS:

Reactive oxygen species

shRNA:

Short hairpin RNA

TRPC1:

Transient receptor potential channel 1

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Acknowledgments

We wish to thank H. Bading for his indispensable help for this project. We also want to thank Alexei Egorov for providing reagents and Jutta Fey for technical assistance in cell culture. This study was supported by a grant from the German Research Foundation (SFB636/A5).

Competing interests

There are no competing interests in regards to this manuscript.

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Authors and Affiliations

  1. Department of Neurology, Harvard Medical School, Massachusetts General Hospital, Charlestown, MA, 02129-4404, USA

    K. Lakshmi Narayanan

  2. Institute of Anatomy and Cell Biology, Department of Molecular Embryology, University of Freiburg, 79104, Freiburg, Germany

    Klaus Unsicker

  3. Department of Neuroscience, The Scripps Research Institute, 130 Scripps way #3C2, Jupiter, FL, 33458, USA

    Srinivasa Subramaniam

  4. Department of Neurobiology, Interdisciplinary Centre for Neurosciences (IZN), University of Heidelberg, 69120, Heidelberg, Germany

    C. Peter Bengston

  5. Department of Medicine, Weill Medical College of Cornell University, New York, NY, 10065, USA

    Krithi Irmady

  6. Institute of Anatomy and Cell Biology, Universitätsmedizin Greifswald, 17487, Greifswald, Germany

    Oliver von Bohlen und Halbach

Authors
  1. K. Lakshmi Narayanan
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  2. Srinivasa Subramaniam
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  3. C. Peter Bengston
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  4. Krithi Irmady
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  5. Klaus Unsicker
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  6. Oliver von Bohlen und Halbach
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Corresponding author

Correspondence to Oliver von Bohlen und Halbach.

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Supplementary Fig. 1

Quantification of TRPC1 mRNA levels normalized to beta actin mRNA levels on the treated cells (n = 3). TRPC1-shRNA significantly reduces TRPC1 mRNA levels (determined by ANOVA followed by Newman–Keuls multiple comparison test). (JPEG 25 kb)

High resolution image (TIFF 250 kb)

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Narayanan, K.L., Subramaniam, S., Bengston, C.P. et al. Role of Transient Receptor Potential Channel 1 (TRPC1) in Glutamate-Induced Cell Death in the Hippocampal Cell Line HT22. J Mol Neurosci 52, 425–433 (2014). https://doi.org/10.1007/s12031-013-0171-9

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  • DOI: https://doi.org/10.1007/s12031-013-0171-9

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