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Two-color, two-photon uncaging of glutamate and GABA

Nature Methods volume 7, pages 123–125 (2010)Cite this article

Abstract

We developed a caged GABA (γ-aminobutyric acid), which, when combined with an appropriate caged glutamate, allows bimodal control of neuronal membrane potential with subcellular resolution using optically independent two-photon uncaging of each neurotransmitter. We used two-color, two-photon uncaging to fire and block action potentials from rat hippocampal CA1 neurons in brain slices with 720-nm and 830-nm light, respectively. Our method should be generalizable to other chemical messenger pairs.

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Figure 1: Absorption spectra, structures and wavelength dependence of evoked currents from two-photon photolysis of CNDI-Glu and N-DCAC-GABA.
Figure 2: Optical, independent two-color, two-photon mapping of functional AMPA and GABA-A receptors on a CA1 pyramidal neuron.
Figure 3: Two-color, two-photon uncaging of glutamate and GABA fired and blocked action potentials.

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Acknowledgements

This work was supported by the US National Institutes of Health grant GM53395 (to G.C.R.E.-D.) and by Grants-in-Aids for Specially Promoted Area number 2000009 (to H.K.), for Scientific Research on Priority Areas number 20021008 (to M.M.) and for Young Scientist (A) number 19680020 (to M.M.) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and Precursory Research for Embryonic Science and Technology and Core Research for Evolutional Science and Technology, Japan Science and Technology Agency (to M.M.). In 1993, G.C.R.E.-D. discussed the idea of two-color uncaging with the late Larry Katz, who had just described the first example of glutamate uncaging in brain slices. With advent of nonlinear methods, we have realized that idea.

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Author notes

  1. Srinivas Kantevari and Masanori Matsuzaki: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA

    Srinivas Kantevari & Graham C R Ellis-Davies

  2. Laboratory of Structural Physiology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine and Center for NanoBio Integration, University of Tokyo, Tokyo, Japan

    Masanori Matsuzaki, Yuya Kanemoto & Haruo Kasai

Authors
  1. Srinivas Kantevari
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  2. Masanori Matsuzaki
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  3. Yuya Kanemoto
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  4. Haruo Kasai
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  5. Graham C R Ellis-Davies
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Contributions

G.C.R.E.-D. and S.K. made and characterized the caged compounds. H.K. designed the three-beam, two-photon microscope and theoretical treatment of two-color uncaging. M.M. and Y.K. did the biological experiments and analyzed the data. G.C.R.E.-D. and H.K. wrote the paper. All authors commented on the manuscript.

Corresponding authors

Correspondence to Haruo Kasai or Graham C R Ellis-Davies.

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Competing interests

G.C.R.E.-D. has filed a patent for dinitroindolinyl caged compounds.

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Kantevari, S., Matsuzaki, M., Kanemoto, Y. et al. Two-color, two-photon uncaging of glutamate and GABA. Nat Methods 7, 123–125 (2010). https://doi.org/10.1038/nmeth.1413

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