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TRPC channels promote cerebellar granule neuron survival

Nature Neuroscience volume 10, pages 559–567 (2007)Cite this article

Abstract

Channels formed by the transient receptor potential (TRP) family of proteins have a variety of physiological functions. Here we report that two members of the TRP cation channel (TRPC) subfamily, TRPC3 and 6, protected cerebellar granule neurons (CGNs) against serum deprivation–induced cell death in cultures and promoted CGN survival in rat brain. In CGN cultures, blocking TRPC channels or downregulating TRPC3 or 6 suppressed brain-derived neurotrophic factor (BDNF)–mediated protection, BDNF-triggered intracellular Ca2+ elevation and BDNF-induced CREB activation. By contrast, overexpressing TRPC3 or 6 increased CREB-dependent reporter gene transcription and prevented apoptosis in the neurons deprived of serum, and this protection was blocked by the dominant negative form of CREB. Furthermore, downregulating TRPC3 or 6 induced CGN apoptosis in neonatal rat cerebellum, and this effect was rescued by overexpressing either TRPC3 or 6. Thus, our findings provide in vitro and in vivo evidence that TRPC channels are important in promoting neuronal survival.

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Figure 1: TRPC3 and 6 are required for BDNF's neuroprotective effect.
Figure 2: PLC and IP3R are necessary for the BDNF neuroprotection.
Figure 3: TRPC3 and 6 contribute to BDNF-triggered [Ca2+]i elevation in CGNs' soma.
Figure 4: TRPC3 and 6 are required for BDNF-induced CREB and ERK activation.
Figure 5: Activation of CREB is necessary for TRPC3 and 6 neuroprotective effects.
Figure 6: Expression pattern of TRPC channels in developing cerebellum and establishment of in vivo electroporation in neonatal rat cerebellum.
Figure 7: TRPC3 and 6 channels promote CGN survival in vivo.

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Acknowledgements

We thank J.W. Putney, Jr., T. Gudermann, D.J. Linden, M.E. Greenberg, L. Li and Z.J. Cheng for providing constructs, C. Montell for TRPC3 antibody, Y.Q. Ding for pCAGGS-GFP, M.M. Poo for comments and Q. Hu for confocal imaging. This work was supported by grants from the 973 Program (2006CB806600) and 30225025, 30621062, U0632006 from National Natural Science Foundation of China.

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Author notes

  1. Yichang Jia and Jian Zhou: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Neural Signal Transduction, Institute of Neuroscience, Shanghai Institutes of Biological Sciences, Key Laboratory of Neurobiology, Shanghai, 200031, China

    Yichang Jia, Jian Zhou, Yilin Tai & Yizheng Wang

  2. The Graduate School, Chinese Academy of Science, 320 Yue Yang Road, Shanghai, 200031, China

    Yichang Jia, Jian Zhou & Yilin Tai

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  1. Yichang Jia
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Contributions

Y.J. and J.Z. conducted the experiments and wrote the manuscript. Y.T. prepared some RNAi constructs, and Y.W. supervised the project and wrote the manuscript.

Corresponding author

Correspondence to Yizheng Wang.

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Supplementary information

Supplementary Fig. 1

Effect of a putative inhibitor of TRPC channels, SKF96365, on BDNF protective effect on cultured CGNs. (PDF 2229 kb)

Supplementary Fig. 2

Transfection efficiency of chemically synthesized siRNA and plasmid on cultured CGNs. (PDF 3899 kb)

Supplementary Fig. 3

An IP3R antagonist, 2-aminoethoxydiphenyl borate (2ApB), suppressed BDNF-dependent neuroprotective effect. (PDF 3491 kb)

Supplementary Fig. 4

Quantitative analysis of the peak ΔR/R stimulated by BDNF together with EGTA. (PDF 721 kb)

Supplementary Fig. 5

The surface expression of TRPC3 and 6 is not altered by BDNF. (PDF 1302 kb)

Supplementary Fig. 6

Effects of SKF96365 on CRE-luciferase activity stimulated by BDNF. (PDF 458 kb)

Supplementary Fig. 7

Wild-type form of human TRPC3 or mouse TRPC6 (pCAGGS-TRPC3/6-IRES-GFP) can be expressed in cultured CGNs. (PDF 773 kb)

Supplementary Fig. 8

Quantitative analysis of phosphorylation of CREB and ERK shown in Figure 5c. (PDF 437 kb)

Supplementary Fig. 9

In vivo electroporation in rat developing cerebellum. (PDF 7082 kb)

Supplementary Fig. 10

The proportion of GFP-positive cells in different layers of rat cerebella as shown in Figure 7b. (PDF 2066 kb)

Supplementary Fig. 11

Schematic diagram depicting a working model for TRPCs neuroprotective effect. (PDF 930 kb)

Supplementary Fig. 12

Effect of downregulation of TRPC3 and 6 on neuronal protective effect of BDNF and IGF-1. (PDF 625 kb)

Supplementary Table 1

siRNA sequences of rat TRPC1, 3, 6 and PLCγ-1. (PDF 589 kb)

Supplementary Methods (PDF 475 kb)

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Jia, Y., Zhou, J., Tai, Y. et al. TRPC channels promote cerebellar granule neuron survival. Nat Neurosci 10, 559–567 (2007). https://doi.org/10.1038/nn1870

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