The patch-clamp technique allows investigation of the electrical excitability of neurons and the functional properties and densities of ion channels. Most patch-clamp recordings from neurons have been made from the soma, the largest structure of individual neurons, while their dendrites, which form the majority of the surface area and receive most of the synaptic input, have been relatively neglected. This protocol describes techniques for recording from the dendrites of neurons in brain slices under direct visual control. Although the basic technique is similar to that used for somatic patching, we describe refinements and optimizations of slice quality, microscope optics, setup stability and electrode approach that are required for maximizing the success rate for dendritic recordings. Using this approach, all configurations of the patch-clamp technique (cell-attached, inside-out, whole-cell, outside-out and perforated patch) can be achieved, even for relatively distal dendrites, and simultaneous multiple-electrode dendritic recordings are also possible. The protocol—from the beginning of slice preparation to the end of the first successful recording—can be completed in 3 h.
Note: In the version of this article initially published online: P. 1235, left column, last line: Quotation marks were misplaced. The sentence should begin: “Although some early, ‘laborious’ efforts..." P. 1236, right column, last four lines, and p. 1238, first text line: References were inserted in the wrong place and misspelled. The sentences should read: “Alternative principal anions are gluconate and methanesulfonate; note that all internal solutions are associated with washout of intracellular factors and some may also have pharmacological effects45 (also see Kaczorowski, C.C., Disterhoft, J.F. & Spruston, N. Soc. Neurosci. Abst. 31, 737.17, 2005). A fluorescent dye (e.g., 1–25 μM Alexa 594) can be included…” P. 1238, first line under EQUIPMENT SETUP: “Recording” was omitted. The sentence should read: “An illustration of a typical setup used for dendritic patch-clamp recording is shown in Figure 1.” P. 1239, Table 1, first item in right column: Mispunctuated. The sentence should read: “Cut sagittally, as parallel to the midline of the cerebellum as possible, on either side of the cerebellar vermis.” P. 1240, last paragraph in Step 11: Text was misplaced. The sentences should read: “It is possible, however, to follow a dendrite deep into the slice from the soma to a distal, more superficial location. Even if the dendrite seems to disappear at points it is possible to spot the same dendrite again at a more distal location. P. 1246, Table 2, last item in right column: Punctuation was misplaced. The sentence should read: “Minimize slice swelling (see above) and rig vibration, and make manipulators as smooth and stable as possible.” These errors have been corrected in all versions of the article.
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Llinas, R. & Nicholson, C. Electrophysiological properties of dendrites and somata in alligator Purkinje cells. J. Neurophysiol. 34, 532–551 (1971).
Wong, R.K., Prince, D.A. & Basbaum, A.I. Intradendritic recordings from hippocampal neurons. Proc. Natl. Acad. Sci. USA 76, 986–990 (1979).
Llinas, R. & Sugimori, M. Electrophysiological properties of in vitro Purkinje cell dendrites in mammalian cerebellar slices. J. Physiol. 305, 197–213 (1980).
Stuart, G.J., Dodt, H.U. & Sakmann, B. Patch-clamp recordings from the soma and dendrites of neurons in brain slices using infrared video microscopy. Pflugers Arch. 423, 511–518 (1993).
Stuart, G. & Häusser, M. Initiation and spread of sodium action potentials in cerebellar Purkinje cells. Neuron 13, 703–712 (1994).
Stuart, G.J. & Sakmann, B. Active propagation of somatic action potentials into neocortical pyramidal cell dendrites. Nature 367, 69–72 (1994).
Magee, J.C. & Johnston, D. Characterization of single voltage-gated Na+ and Ca2+ channels in apical dendrites of rat CA1 pyramidal neurons. J. Physiol. 487 (Pt 1): 67–90 (1995).
Magee, J.C. Dendritic hyperpolarization-activated currents modify the integrative properties of hippocampal CA1 pyramidal neurons. J. Neurosci. 18, 7613–7624 (1998).
Bekkers, J.M. Distribution and activation of voltage-gated potassium channels in cell-attached and outside-out patches from large layer 5 cortical pyramidal neurons of the rat. J. Physiol. 525 Pt 3: 611–620 (2000).
Korngreen, A. & Sakmann, B. Voltage-gated K+ channels in layer 5 neocortical pyramidal neurones from young rats: subtypes and gradients. J. Physiol. 525 Pt 3: 621–639 (2000).
Schaefer, A.T., Helmstaedter, M., Sakmann, B. & Korngreen, A. Correction of conductance measurements in non-space-clamped structures: 1. Voltage-gated K+ channels. Biophys. J. 84, 3508–3528 (2003).
Hoffman, D.A., Magee, J.C., Colbert, C.M. & Johnston, D. K+ channel regulation of signal propagation in dendrites of hippocampal pyramidal neurons. Nature 387, 869–875 (1997).
Colbert, C.M., Magee, J.C., Hoffman, D.A. & Johnston, D. Slow recovery from inactivation of Na+ channels underlies the activity-dependent attenuation of dendritic action potentials in hippocampal CA1 pyramidal neurons. J. Neurosci. 17, 6512–6521 (1997).
Martina, M., Yao, G.L. & Bean, B.P. Properties and functional role of voltage-dependent potassium channels in dendrites of rat cerebellar Purkinje neurons. J. Neurosci. 23, 5698–5707 (2003).
Poolos, N.P. & Johnston, D. Calcium-activated potassium conductances contribute to action potential repolarization at the soma but not the dendrites of hippocampal CA1 pyramidal neurons. J. Neurosci. 19, 5205–5212 (1999).
Roth, A. & Häusser, M. Compartmental models of rat cerebellar Purkinje cells based on simultaneous somatic and dendritic patch-clamp recordings. J. Physiol. 535, 445–472 (2001).
Stuart, G. & Spruston, N. Determinants of voltage attenuation in neocortical pyramidal neuron dendrites. J. Neurosci. 18, 3501–3510 (1998).
Golding, N.L., Mickus, T.J., Katz, Y., Kath, W.L. & Spruston, N. Factors mediating powerful voltage attenuation along CA1 pyramidal neuron dendrites. J. Physiol. 568, 69–82 (2005).
Larkum, M.E., Zhu, J.J. & Sakmann, B. Dendritic mechanisms underlying the coupling of the dendritic with the axonal action potential initiation zone of adult rat layer 5 pyramidal neurons. J. Physiol. 533, 447–466 (2001).
Williams, S.R. Spatial compartmentalization and functional impact of conductance in pyramidal neurons. Nat. Neurosci. 7, 961–967 (2004).
Gasparini, S., Migliore, M. & Magee, J.C. On the initiation and propagation of dendritic spikes in CA1 pyramidal neurons. J. Neurosci. 24, 11046–11056 (2004).
Pouille, F. & Scanziani, M. Enforcement of temporal fidelity in pyramidal cells by somatic feed-forward inhibition. Science 293, 1159–1163 (2001).
London, M. & Häusser, M. Dendritic computation. Annu. Rev. Neurosci. 28, 503–532 (2005).
Gulledge, A.T., Kampa, B.M. & Stuart, G.J. Synaptic integration in dendritic trees. J. Neurobiol. 64, 75–90 (2005).
Migliore, M. & Shepherd, G.M. Emerging rules for the distributions of active dendritic conductances. Nat. Rev. Neurosci. 3, 362–370 (2002).
Häusser, M., Spruston, N. & Stuart, G.J. Diversity and dynamics of dendritic signaling. Science 290, 739–744 (2000).
Magee, J.C. & Johnston, D. Plasticity of dendritic function. Curr. Opin. Neurobiol. 15, 334–342 (2005).
Edwards, F.A., Konnerth, A., Sakmann, B. & Takahashi, T. A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system. Pflugers Arch. 414, 600–612 (1989).
Usowicz, M.M., Sugimori, M., Cherksey, B. & Llinas, R. P-type calcium channels in the somata and dendrites of adult cerebellar Purkinje cells. Neuron 9, 1185–1199 (1992).
Huguenard, J.R., Hamill, O.P. & Prince, D.A. Sodium channels in dendrites of rat cortical pyramidal neurons. Proc. Natl. Acad. Sci. USA 86, 2473–2477 (1989).
Dodt, H.U., Frick, A., Kampe, K. & Zieglgansberger, W. NMDA and AMPA receptors on neocortical neurons are differentially distributed. Eur. J. Neurosci. 10, 3351–3357 (1998).
Williams, S.R. & Stuart, G.J. Dependence of EPSP efficacy on synapse location in neocortical pyramidal neurons. Science 295, 1907–1910 (2002).
Häusser, M., Stuart, G., Racca, C. & Sakmann, B. Axonal initiation and active dendritic propagation of action potentials in substantia nigra neurons. Neuron 15, 637–647 (1995).
Martina, M., Schultz, J.H., Ehmke, H., Monyer, H. & Jonas, P. Functional and molecular differences between voltage-gated K+ channels of fast-spiking interneurons and pyramidal neurons of rat hippocampus. J. Neurosci. 18, 8111–8125 (1998).
Williams, S.R. & Stuart, G.J. Action potential backpropagation and somato-dendritic distribution of ion channels in thalamocortical neurons. J. Neurosci. 20, 1307–1317 (2000).
Larkum, M.E., Rioult, M.G. & Luscher, H.R. Propagation of action potentials in the dendrites of neurons from rat spinal cord slice cultures. J. Neurophysiol. 75, 154–170 (1996).
Geiger, J.R. & Jonas, P. Dynamic control of presynaptic Ca(2+) inflow by fast-inactivating K(+) channels in hippocampal mossy fiber boutons. Neuron 28, 927–939 (2000).
Forsythe, I.D. Direct patch recording from identified presynaptic terminals mediating glutamatergic EPSCs in the rat CNS, in vitro. J. Physiol. 479 (Pt 3): 381–387 (1994).
Southan, A.P. & Robertson, B. Patch-clamp recordings from cerebellar basket cell bodies and their presynaptic terminals reveal an asymmetric distribution of voltage-gated potassium channels. J. Neurosci. 18, 948–955 (1998).
Gulledge, A.T. & Stuart, G.J. Excitatory actions of GABA in the cortex. Neuron 37, 299–309 (2003).
Stuart, G. Patch-pipet recording in brain slices. Curr. Prot. Neurosci. 6, 1–10 (1998).
Blanton, M.G., Lo Turco, J.J. & Kriegstein, A.R. Whole cell recording from neurons in slices of reptilian and mammalian cerebral cortex. J. Neurosci. Methods 30, 203–210 (1989).
Geiger, J.R. et al. Patch-clamp recording in brain slices with improved slicer technology. Pflugers Arch. 443, 491–501 (2002).
Aghajanian, G.K. & Rasmussen, K. Intracellular studies in the facial nucleus illustrating a simple new method for obtaining viable motoneurons in adult rat brain slices. Synapse 3, 331–338 (1989).
Velumian, A.A., Zhang, L., Pennefather, P. & Carlen, P.L. Reversible inhibition of IK, IAHP, Ih and ICa currents by internally applied gluconate in rat hippocampal pyramidal neurones. Pflugers Arch 433, 343–450 (1997).
Hamill, O.P., Marty, A., Neher, E., Sakmann, B. & Sigworth, F.J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391, 85–100 (1981).
Frick, A., Magee, J. & Johnston, D. LTP is accompanied by an enhanced local excitability of pyramidal neuron dendrites. Nat. Neurosci. 7, 126–135 (2004).
Frick, A. & Johnston, D. Plasticity of dendritic excitability. J. Neurobiol. 64, 100–115 (2005).
Dodt, H.U., Eder, M., Schierloh, A. & Zieglgansberger, W. Infrared-guided laser stimulation of neurons in brain slices. Sci. STKE, PL 2(2002).
Dodt, H.U. & Zieglgansberger, W. Visualizing unstained neurons in living brain slices by infrared DIC-videomicroscopy. Brain Res. 537, 333–336 (1990).
Lanni, F. & Keller, H.E. Microscopy and Microscope Optical Systems (Eds. Yuste, R., Lanni, F. & Konnerth, A.) Cold Spring Harbor Laboratory Press, 1999.
Foskett, J.K. Simultaneous Nomarski and fluorescence imaging during video microscopy of cells. Am. J. Physiol. 255, C566–C571 (1988).
We thank M. London, F. Pouille, M. Scanziani and J. Sjöström for their helpful comments on the manuscript. This work was supported by the Gatsby Foundation (M.H.), the Wellcome Trust (M.H., J.T.D, E.A.R.), the NH&MRC of Australia (G.J.S., M.H.P.K., J.J.L.) and the US National Institutes of Health (NS-35180, NS-46064 to N.S.).
The authors declare no competing financial interests.
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