Targeted intracellular voltage recordings from dendritic spines using quantum-dot-coated nanopipettes

Abstract

Dendritic spines are the primary site of excitatory synaptic input onto neurons, and are biochemically isolated from the parent dendritic shaft by their thin neck. However, due to the lack of direct electrical recordings from spines, the influence that the neck resistance has on synaptic transmission, and the extent to which spines compartmentalize voltage, specifically excitatory postsynaptic potentials, albeit critical, remains controversial. Here, we use quantum-dot-coated nanopipette electrodes (tip diameters 15–30 nm) to establish the first intracellular recordings from targeted spine heads under two-photon visualization. Using simultaneous somato-spine electrical recordings, we find that back propagating action potentials fully invade spines, that excitatory postsynaptic potentials are large in the spine head (mean 26 mV) but are strongly attenuated at the soma (0.5–1 mV) and that the estimated neck resistance (mean 420 MΩ) is large enough to generate significant voltage compartmentalization. Nanopipettes can thus be used to electrically probe biological nanostructures.

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Figure 1: Experimental overview.
Figure 2: The nanopipette membrane interface.
Figure 3: Nanopipette recordings from cultures.
Figure 4: Nanopipette recordings from neocortical slices.
Figure 5: Nanopipette recordings in spines reveals electrical compartmentalization.

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Acknowledgements

K.J. would like to thank M. Barbic of the applied physics and instrumentation group at Janelia Farms for initial assistance with initial SEM imaging. This work was supported by the National Institute of Mental Health (NIMH) (R01MH101218, R01MH100561) and the Kavli Institute of Brain Science. This material is also based on work supported by, or in part by, the US Army Research Laboratory and the US Army Research Office under contract number W911NF-12-1-0594 (MURI).

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K.J. and R.Y. designed and conceptualized the study. J.J.H. performed slice preparation and provided technical support. I.J.-L.P., W.D.A.M.D.B. and J.S.O. synthesized, characterized and provided quantum dots. D.M.T. and K.J. performed SEM imaging. A.S. prepared cell cultures. D.S.P. provided technical support. O.S, K.L.S. and R.Y. advised on experiments. R.Y. supervised the overall study. K.J. performed all the experiments, modelling and analysis and wrote the manuscript. All the authors discussed the manuscript and provided comments.

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Correspondence to Krishna Jayant.

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The authors declare no competing financial interests.

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Jayant, K., Hirtz, J., Plante, I. et al. Targeted intracellular voltage recordings from dendritic spines using quantum-dot-coated nanopipettes. Nature Nanotech 12, 335–342 (2017). https://doi.org/10.1038/nnano.2016.268

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