Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Ultra light-sensitive and fast neuronal activation with the Ca2+-permeable channelrhodopsin CatCh

Nature Neuroscience volume 14, pages 513–518 (2011)Cite this article

Subjects

Abstract

The light-gated cation channel channelrhodopsin-2 (ChR2) has rapidly become an important tool in neuroscience, and its use is being considered in therapeutic interventions. Although wild-type and known variant ChR2s are able to drive light-activated spike trains, their use in potential clinical applications is limited by either low light sensitivity or slow channel kinetics. We present a new variant, calcium translocating channelrhodopsin (CatCh), which mediates an accelerated response time and a voltage response that is ∼70-fold more light sensitive than that of wild-type ChR2. CatCh's superior properties stem from its enhanced Ca2+ permeability. An increase in [Ca2+]i elevates the internal surface potential, facilitating activation of voltage-gated Na+ channels and indirectly increasing light sensitivity. Repolarization following light-stimulation is markedly accelerated by Ca2+-dependent BK channel activation. Our results demonstrate a previously unknown principle: shifting permeability from monovalent to divalent cations to increase sensitivity without compromising fast kinetics of neuronal activation. This paves the way for clinical use of light-gated channels.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Homology model of ChR2 based on the sensory rhodopsin 2 structure (PDB accession number 1H2S).
Figure 2: Biophysical characterization of CatCh in HEK293 cells and Xenopus oocytes.
Figure 3: CatCh expression in hippocampal cultured neurons.
Figure 4: Fast and high-sensitivity neural photostimulation.

References

  1. Nagel, G. et al. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc. Natl. Acad. Sci. USA 100, 13940–13945 (2003).

    Article  CAS  Google Scholar 

  2. Nagel, G. et al. Light activation of channelrhodopsin-2 in excitable cells of Caenorhabditis elegans triggers rapid behavioral responses. Curr. Biol. 15, 2279–2284 (2005).

    Article  CAS  Google Scholar 

  3. Boyden, E.S., Zhang, F., Bamberg, E., Nagel, G. & Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat. Neurosci. 8, 1263–1268 (2005).

    Article  CAS  PubMed  Google Scholar 

  4. Zhang, F. et al. Multimodal fast optical interrogation of neural circuitry. Nature 446, 633–639 (2007).

    Article  CAS  PubMed  Google Scholar 

  5. Bamann, C., Gueta, R., Kleinlogel, S., Nagel, G. & Bamberg, E. Structural guidance of the photocycle of channelrhodopsin-2 by an interhelical hydrogen bond. Biochemistry 49, 267–278 (2010).

    Article  CAS  PubMed  Google Scholar 

  6. Berndt, A., Yizhar, O., Gunaydin, L., Hegemann, P. & Deisseroth, K. Bi-stable neural state switches. Nat. Neurosci. 12, 229–234 (2009).

    Article  CAS  Google Scholar 

  7. Lin, J.Y., Lin, M., Steinbach, P. & Tsien, R. Characterization of engineered channelrhodopsin variants with improved properties and kinetics. Biophys. J. 96, 1803–1814 (2009).

    Article  CAS  PubMed  Google Scholar 

  8. Lagali, P.S. et al. Light-activated channels targeted to ON bipolar cells restore visual function in retinal degeneration. Nat. Neurosci. 11, 667–675 (2008).

    Article  CAS  Google Scholar 

  9. Bi, A. et al. Ectopic expression of a microbial-type rhodopsin restores visual responses in mice with photoreceptor degeneration. Neuron 50, 23–33 (2006).

    Article  CAS  PubMed  Google Scholar 

  10. Frankenhaeuser, B. & Hodgkin, A. The action of calcium on the electrical properties of squid axons. J. Physiol. (Lond.) 137, 218–244 (1957).

    Article  CAS  Google Scholar 

  11. Hille, B. Ion Channels of Excitable Membranes 3rd edn., 649–662 (Sinauer, Sunderland, Massachusetts, USA, 2001).

  12. Gunaydin, L.A. et al. Ultrafast optogenetic control. Nat. Neurosci. 13, 387–392 (2010).

    Article  CAS  PubMed  Google Scholar 

  13. Bamann, C., Kirsch, T., Nagel, G. & Bamberg, E. Spectral characteristics of the photocycle of channelrhodopsin-2 and its implication for channel function. J. Mol. Biol. 375, 686–694 (2008).

    Article  CAS  PubMed  Google Scholar 

  14. Feldbauer, K. et al. Channelrhodopsin-2 is a leaky proton pump. Proc. Natl. Acad. Sci. USA 106, 12317–12322 (2009).

    Article  CAS  PubMed  Google Scholar 

  15. Caldwell, J. et al. Increases in intracellular calcium triggered by channelrhodopsin-2 potentiate the response of metabotropic glutamate receptor mGluR7. J. Biol. Chem. 283, 24300–24307 (2008).

    Article  CAS  PubMed  Google Scholar 

  16. Weber, W. Ion currents in Xenopus laevis oocytes: state of the art. Biochim. Biophys. Acta 1421, 213–233 (1999).

    Article  CAS  PubMed  Google Scholar 

  17. Thyagarajan, S. et al. Visual function in mice with photoreceptor degeneration and transgenic expression of channelrhodopsin 2 in ganglion cells. J. Neurosci. 30, 8745–8758 (2010).

    Article  CAS  PubMed  Google Scholar 

  18. Hille, B., Woodhull, B. & Shapiro, B. Negative surface charge near sodium channels of nerve: divalent ions, monovalent ions, and pH. Phil. Trans. R. Soc. Lond. B 270, 301–318 (1975).

    Article  CAS  Google Scholar 

  19. Muller, R.U. & Finkelstein, A. The effect of surface charge on the voltage-dependent conductance induced in thin lipid membranes by monazomycin. J. Gen. Physiol. 60, 285–306 (1972).

    Article  CAS  PubMed  Google Scholar 

  20. Faber, E.S. & Sah, P. Calcium-activated potassium-channels: multiple contributions to neuronal function. Neuroscientist 9, 181–194 (2003).

    Article  CAS  PubMed  Google Scholar 

  21. Joh, N.H., Oberai, A., Yang, D., Whitelegge, J. & Bowie, J. Similar energetic contributions of packing in the core of membrane and water-soluble proteins. J. Am. Chem. Soc. 131, 10846–10847 (2009).

    Article  CAS  PubMed  Google Scholar 

  22. Subramaniam, S., Faruqi, A., Oesterhelt, D. & Henderson, R. Electron diffraction studies of light-induced conformational changes in the Leu-93→Ala bacteriorhodopsin mutant. Proc. Natl. Acad. Sci. USA 94, 1767–1772 (1997).

    Article  CAS  PubMed  Google Scholar 

  23. Subramaniam, S., Greenhalgh, D., Rath, P., Rothschild, K. & Khorana, H. Replacement of leucine-93 by alanine or threonine slows down the decay of the N and O intermediates in the photocycle of bacteriorhodopsin: implications for proton uptake and 13-cis-retinal all-trans-retinal reisomerization. Proc. Natl. Acad. Sci. USA 88, 6873–6877 (1991).

    Article  CAS  PubMed  Google Scholar 

  24. Nack, M. et al. The DC gate in Channelrhodopsin-2: crucial hydrogen bonding interaction between C128 and D156. Photochem. Photobiol. Sci. 9, 194–198 (2010).

    Article  CAS  PubMed  Google Scholar 

  25. Cady, C., Evans, M.S. & Brewer, G.J. Age-related differences in NMDA responses in cultured rat hippocampal neurons. Brain Res. 921, 1–11 (2001).

    Article  CAS  PubMed  Google Scholar 

  26. Busskamp, V. et al. Genetic reactivation of cone photoreceptors restores visual responses in retinitis pigmentosa. Science 329, 413–417 (2010).

    Article  CAS  Google Scholar 

  27. Humphrey, W., Dalke, A. & Schulten, K. VMD: visual molecular dynamics. J. Mol. Graph. 14, 33–38 (1996).

    Article  CAS  PubMed  Google Scholar 

  28. Lorenz, C., Pusch, M. & Jentsch, T. Heteromultimeric ClC chloride channels with novel properties. Proc. Natl. Acad. Sci. USA 93, 13362–13366 (1996).

    Article  CAS  PubMed  Google Scholar 

  29. Allocca, M. et al. Novel adeno-associated virus serotypes efficiently transduce murine photoreceptors. J. Virol. 81, 11372–11380 (2007).

    Article  CAS  PubMed  Google Scholar 

  30. de Felipe, P. et al. E unum pluribus: multiple proteins from a self-processing polyprotein. Trends Biotechnol. 24, 68–75 (2006).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to thank I. Bartnik for the preparation of the hippocampal neuron cultures, S. O'Shea for the help with the calcium imaging experiments, V. Busskamp for the support in recombinant adeno-associated virus construction, H. Biehl for excellent technical assistance, and K. Hartung, H. Spors, U. Terpitz and M. van Wyk for helpful discussions. The work was supported by grants from the Deutsche Forschungsgemeinschaft Sonderforschungsbereich 807, Centre of Excellence Frankfurt Macromolecular Complexes and the Federal Ministry of Education and Research of Germany (01GQ0815) to E.B., and by the Max Planck Society.

Author information

Author notes

  1. Robert E Dempski

    Present address: Present address: Department of Chemistry and Biochemistry, Worcester Polytechnic Institute, Worcester, Massachusetts, USA.,

  2. Katrin Feldbauer and Robert E Dempski: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biophysical Chemistry, Max Planck Institute of Biophysics, Frankfurt am Main, Germany

    Sonja Kleinlogel, Katrin Feldbauer, Robert E Dempski, Heike Fotis, Phillip G Wood, Christian Bamann & Ernst Bamberg

  2. Chemical and Pharmaceutical Sciences Department, Johann-Wolfgang-Goethe-University, Frankfurt am Main, Germany

    Ernst Bamberg

Authors
  1. Sonja Kleinlogel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Katrin Feldbauer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Robert E Dempski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heike Fotis
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Phillip G Wood
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christian Bamann
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ernst Bamberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

S.K., R.E.D., P.G.W. and E.B. conceived the experiments. S.K., K.F., H.F. and C.B. carried out the experiments. S.K., C.B. and K.F. performed the data analysis. S.K., C.B. and E.B. wrote the manuscript.

Corresponding author

Correspondence to Ernst Bamberg.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3, Supplementary Note and Supplementary Discussion (PDF 251 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kleinlogel, S., Feldbauer, K., Dempski, R. et al. Ultra light-sensitive and fast neuronal activation with the Ca2+-permeable channelrhodopsin CatCh. Nat Neurosci 14, 513–518 (2011). https://doi.org/10.1038/nn.2776

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn.2776

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing