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PLoS One. 2009 Dec 4;4(12):e8185. doi: 10.1371/journal.pone.0008185.

Temporal control of immediate early gene induction by light.

Author information

1
Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.

Abstract

BACKGROUND:

The light-gated cation channel channelrhodopsin-2 (ChR2) is a powerful tool for the optical induction of action potentials in neurons. Mutations of the cysteine 128 (C128) residue have been shown to greatly extend the lifetime of the conducting state of ChR2. However, until now, only subthreshold depolarizations have been reported from C128 mutants.

METHODS AND FINDINGS:

Here we report the induction of long high-frequency spike trains by brief light pulses in ChR2(C128A)-transfected pyramidal cells in hippocampal slice culture. ChR2(C128A)-mediated spike bursts triggered expression of the immediate early gene c-fos in pyramidal neurons. Robust and cell-specific expression of c-Fos protein was detected after a single blue light pulse and depended on action potential firing, but not on synaptic activity. However, photocurrents diminished upon repeated stimulation and limited the number of action potential bursts that could be elicited.

CONCLUSIONS:

We conclude that the C128A mutant is not suitable for chronic stimulation of neurons, but very useful for light-controlled induction of immediate early genes. This property of ChR2(C128A) could be harnessed to control the expression of proteins under control of the c-fos promoter with precise timing and single cell specificity.

PMID:
19997631
PMCID:
PMC2780714
DOI:
10.1371/journal.pone.0008185
[Indexed for MEDLINE]
Free PMC Article

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