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.
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Yuste, R. Electrical compartmentalization in dendritic spines. Annu. Rev. Neurosci. 36, 429–449 (2013).
Grunditz, Å., Holbro, N., Tian, L., Zuo, Y. & Oertner, T. G. Spine neck plasticity controls postsynaptic calcium signals through electrical compartmentalization. J. Neurosci. 28, 13457–13466 (2008).
Hao, J. & Oertner, T. G. Depolarization gates spine calcium transients and spike-timing-dependent potentiation. Curr. Opin. Neurobiol. 22, 509–515 (2012).
Grutzendler, J., Kasthuri, N. & Gan, W.-B. Long-term dendritic spine stability in the adult cortex. Nature 420, 812–816 (2002).
Yang, G., Pan, F. & Gan, W.-B. Stably maintained dendritic spines are associated with lifelong memories. Nature 462, 920–924 (2009).
Yuste, R. & Bonhoeffer, T. Morphological changes in dendritic spines associated with long-term synaptic plasticity. Annu. Rev. Neurosci. 24, 1071–1089 (2001).
Yuste, R. & Denk, W. Dendritic spines as basic functional units of neuronal integration. Nature 375, 682–684 (1995).
Müller, W. & Connor, J. A. Dendritic spines as individual neuronal compartments for synaptic Ca2+ responses. Nature 354, 73–76 (1991).
Bloodgood, B. L. & Sabatini, B. L. Neuronal activity regulates diffusion across the neck of dendritic spines. Science 310, 866–869 (2005).
Harnett, M. T., Makara, J. K., Spruston, N., Kath, W. L. & Magee, J. C. Synaptic amplification by dendritic spines enhances input cooperativity. Nature 491, 599–602 (2012).
Perkel, D. H. & Perkel, D. J. Dendritic spines: role of active membrane in modulating synaptic efficacy. Brain Res. 325, 331–335 (1985).
Shepherd, G. et al. Signal enhancement in distal cortical dendrites by means of interactions between active dendritic spines. Proc. Natl Acad. Sci. USA 82, 2192–2195 (1985).
Miller, J. P., Rall, W. & Rinzel, J. Synaptic amplification by active membrane in dendritic spines. Brain Res. 325, 325–330 (1985).
Bloodgood, B. L., Giessel, A. J. & Sabatini, B. L. Biphasic synaptic Ca influx arising from compartmentalized electrical signals in dendritic spines. PLoS Biol. 7, 1–10 (2009).
Araya, R., Jiang, J., Eisenthal, K. B. & Yuste, R. The spine neck filters membrane potentials. Proc. Natl Acad. Sci. USA 103, 17961–17966 (2006).
Acker, C. D., Yan, P. & Loew, L. M. Single-voxel recording of voltage transients in dendritic spines. Biophys. J. 101, L11–L13 (2011).
Segev, I. & Rall, W. Computational study of an excitable dendritic spine. J. Neurophysiol. 60, 499–523 (1988).
Koch, C. & Zador, A. The function of dendritic spines: devices subserving biochemical rather than electrical computation. J. Neurosci. 13, 413–422 (1993).
Svoboda, K., Tank, D. W. & Denk, W. Direct measurement of coupling between dendritic spines and shafts. Science 272, 716–719 (1996).
Tønnesen, J., Katona, G., Rózsa, B. & Nägerl, U. V. Spine neck plasticity regulates compartmentalization of synapses. Nat. Neurosci. 17, 678–685 (2014).
Palmer, L. M. & Stuart, G. J. Membrane potential changes in dendritic spines during action potentials and synaptic input. J. Neurosci. 29, 6897–6903 (2009).
Hochbaum, D. R. et al. All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins. Nat. Methods 11, 825–833 (2014).
Acker, C. D., Hoyos, E. & Loew, L. M. EPSPs measured in proximal dendritic spines of cortical pyramidal neurons. eneuro 3, 1–13 (2016).
Popovic, M. A., Gao, X., Carnevale, N. T. & Zecevic, D. Cortical dendritic spine heads are not electrically isolated by the spine neck from membrane potential signals in parent dendrites. Cerebral Cortex 24, 385–395 (2012).
Popovic, M. A., Carnevale, N., Rozsa, B. & Zecevic, D. Electrical behaviour of dendritic spines as revealed by voltage imaging. Nat. Commun. 6, 8436 (2015).
Nuriya, M., Jiang, J., Nemet, B., Eisenthal, K. B. & Yuste, R. Imaging membrane potential in dendritic spines. Proc. Natl Acad. Sci. USA 103, 786–790 (2006).
Qing, Q. et al. Free-standing kinked nanowire transistor probes for targeted intracellular recording in three dimensions. Nat. Nanotech. 9, 142–147 (2014).
Qing, Q. et al. Nanowire transistor arrays for mapping neural circuits in acute brain slices. Proc. Natl Acad. Sci. USA 107, 1882–1887 (2010).
Robinson, J. T. et al. Vertical nanowire electrode arrays as a scalable platform for intracellular interfacing to neuronal circuits. Nat. Nanotech. 7, 180–184 (2012).
Tian, B. et al. Three-dimensional, flexible nanoscale field-effect transistors as localized bioprobes. Science 329, 830–834 (2010).
Xie, C., Lin, Z., Hanson, L., Cui, Y. & Cui, B. Intracellular recording of action potentials by nanopillar electroporation. Nat. Nanotech. 7, 185–190 (2012).
Singhal, R. et al. Multifunctional carbon-nanotube cellular endoscopes. Nat. Nanotech. 6, 57–64 (2011).
Novak, P. et al. Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels. Neuron 79, 1067–1077 (2013).
Spira, M. E. & Hai, A. Multi-electrode array technologies for neuroscience and cardiology. Nat. Nanotech. 8, 83–94 (2013).
Fendyur, A. & Spira, M. E. Toward on-chip, in-cell recordings from cultured cardiomyocytes by arrays of gold mushroom-shaped microelectrodes. Front. Neuroeng. 5, 1–10 (2012).
Hai, A., Shappir, J. & Spira, M. E. In-cell recordings by extracellular microelectrodes. Nat. Methods 7, 200–202 (2010).
Angle, M. R. & Schaefer, A. T. Neuronal recordings with solid-conductor intracellular nanoelectrodes (SCINEs). PLoS ONE 7, e43194 (2012).
Alivisatos, A. P. et al. Nanotools for neuroscience and brain activity mapping. ACS Nano 7, 1850–1866 (2013).
Hu, H. & Jonas, P. A supercritical density of Na+ channels ensures fast signaling in GABAergic interneuron axons. Nat. Neurosci. 17, 686–693 (2014).
Stuart, G. J. & Sakmann, B. Active propagation of somatic action potentials into neocortical pyramidal cell dendrites. Nature 367, 69–72 (1994).
Stuart, G. J. & Häusser, M. Dendritic coincidence detection of EPSPs and action potentials. Nat. Neurosci. 4, 63–71 (2001).
Holthoff, K., Zecevic, D. & Konnerth, A. Rapid time course of action potentials in spines and remote dendrites of mouse visual cortex neurons. J. Physiol. 588, 1085–1096 (2010).
Andrasfalvy, B. K. et al. Quantum dot-based multiphoton fluorescent pipettes for targeted neuronal electrophysiology. Nat. Methods 11, 1237–1241 (2014).
Purves, R. Microelectrode Methods for Intracellular Recording and Ionophoresis (Academic, 1981).
Ince, C. et al. Intracellular microelectrode measurements in small cells evaluated with the patch clamp technique. Biophys. J. 50, 1203–1209 (1986).
Brette, R. et al. High-resolution intracellular recordings using a real-time computational model of the electrode. Neuron 59, 379–391 (2008).
Tsay, D. & Yuste, R. On the electrical function of dendritic spines. Trend. Neurosci. 27, 77–83 (2004).
Lin, Z. C., Xie, C., Osakada, Y., Cui, Y. & Cui, B. Iridium oxide nanotube electrodes for sensitive and prolonged intracellular measurement of action potentials. Nat. Commun. 5, 3206 (2014).
Bi, G.-q. & Poo, M.-m. Distributed synaptic modification in neural networks induced by patterned stimulation. Nature 401, 792–796 (1999).
Duan, X. et al. Intracellular recordings of action potentials by an extracellular nanoscale field-effect transistor. Nat. Nanotech. 7, 174–179 (2012).
Fu, T.-M. et al. Sub-10-nm intracellular bioelectronic probes from nanowire–nanotube heterostructures. Proc. Natl Acad. Sci. USA 111, 1259–1264 (2014).
Almquist, B. D. & Melosh, N. A. Fusion of biomimetic stealth probes into lipid bilayer cores. Proc. Natl Acad. Sci. USA 107, 5815–5820 (2010).
Angle, M. R., Cui, B. & Melosh, N. A. Nanotechnology and neurophysiology. Curr. Opin. Neurobiol. 32, 132–140 (2015).
Beaudoin III, G. M. et al. Culturing pyramidal neurons from the early postnatal mouse hippocampus and cortex. Nat. Protoc. 7, 1741–1754 (2012).
Packer, A. M. Understanding the Nervous System as an Information Processing Machine: Dense, Nonspecific, Canonical Microcircuit Architecture of Inhibition in Neocortex and a Neural Circuit for Angular Velocity Computation PhD thesis, Columbia Univ. (2011).
Garaj, S. et al. Graphene as a subnanometre trans-electrode membrane. Nature 467, 190–193 (2010).
Araya, R. Input transformation by dendritic spines of pyramidal neurons. Front. Neuroanatomy, 8, 1–18 (2014).
Denk, W., Yuste, R., Svoboda, K. & Tank, D. W. Imaging calcium dynamics in dendritic spines. Curr. Opin. Neurobiol. 6, 372–378 (1996).
Thevenaz, P., Ruttimann, U. E. & Unser, M. A pyramid approach to subpixel registration based on intensity. IEEE Trans. Image Processing 7, 27–41 (1998).
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).
The authors declare no competing financial interests.
About this article
Cite this article
Jayant, K., Hirtz, J., Plante, IL. 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
Current Pharmacology Reports (2021)
Journal of General Physiology (2021)
Large, Stable Spikes Exhibit Differential Broadening in Excitatory and Inhibitory Neocortical Boutons
Cell Reports (2021)
Angewandte Chemie International Edition (2021)
Science Advances (2021)