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A practical guide to the synthesis of dinitroindolinyl-caged neurotransmitters

Abstract

This protocol describes a method for efficient chemical synthesis of dinitroindolinyl derivatives of glutamate and γ-aminobutyric acid. These caged neurotransmitters are currently the most chemically and photochemically efficient probes for two-photon photolysis in living brain slices. The protocol only requires basic organic synthesis equipment, and no silica gel column chromatography or NMR spectroscopy is needed at any stage. HPLC is used to purify the caged transmitters at the end of the syntheses. Thus, the synthesis of dinitroindolinyl-caged neurotransmitters is within the scope of a modestly equipped chemistry laboratory.

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Figure 1
Figure 2: Scheme for the synthesis of CDNI-Glu and -GABA.
Figure 3: Analytical HPLC of CDNI-Glu synthesis.
Figure 4: Preparative HPLC purification of CDNI-Glu synthesis.
Figure 5: Preparative HPLC purification of CDNI-GABA synthesis.

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Acknowledgements

This work was supported by a grant from the US National Institutes of Health (GM53395) to G.C.R.E.-D.

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G.C.R.E.-D. performed the synthesis and wrote the paper.

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Correspondence to Graham C R Ellis-Davies.

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G.C.R.E.-D. has filed a patent in the United States of America on the synthesis of dinitroindolinyl-caged neurotransmitters.

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Ellis-Davies, G. A practical guide to the synthesis of dinitroindolinyl-caged neurotransmitters. Nat Protoc 6, 314–326 (2011). https://doi.org/10.1038/nprot.2010.193

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