Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Dendritic patch-clamp recording

This article has been updated

Abstract

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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Patch-clamp setup.
Figure 2: Imaging dendrites using videomicroscopy.
Figure 3: Steps in dendritic patching.

Change history

  • 30 November 2006

    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.

References

  1. 1

    Llinas, R. & Nicholson, C. Electrophysiological properties of dendrites and somata in alligator Purkinje cells. J. Neurophysiol. 34, 532–551 (1971).

    CAS  Article  Google Scholar 

  2. 2

    Wong, R.K., Prince, D.A. & Basbaum, A.I. Intradendritic recordings from hippocampal neurons. Proc. Natl. Acad. Sci. USA 76, 986–990 (1979).

    CAS  Article  Google Scholar 

  3. 3

    Llinas, R. & Sugimori, M. Electrophysiological properties of in vitro Purkinje cell dendrites in mammalian cerebellar slices. J. Physiol. 305, 197–213 (1980).

    CAS  Article  Google Scholar 

  4. 4

    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).

    CAS  Article  Google Scholar 

  5. 5

    Stuart, G. & Häusser, M. Initiation and spread of sodium action potentials in cerebellar Purkinje cells. Neuron 13, 703–712 (1994).

    CAS  Article  Google Scholar 

  6. 6

    Stuart, G.J. & Sakmann, B. Active propagation of somatic action potentials into neocortical pyramidal cell dendrites. Nature 367, 69–72 (1994).

    CAS  Article  Google Scholar 

  7. 7

    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).

    CAS  Article  Google Scholar 

  8. 8

    Magee, J.C. Dendritic hyperpolarization-activated currents modify the integrative properties of hippocampal CA1 pyramidal neurons. J. Neurosci. 18, 7613–7624 (1998).

    CAS  Article  Google Scholar 

  9. 9

    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).

    CAS  Article  Google Scholar 

  10. 10

    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).

    CAS  Article  Google Scholar 

  11. 11

    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).

    CAS  Article  Google Scholar 

  12. 12

    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).

    CAS  Article  Google Scholar 

  13. 13

    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).

    CAS  Article  Google Scholar 

  14. 14

    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).

    CAS  Article  Google Scholar 

  15. 15

    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).

    CAS  Article  Google Scholar 

  16. 16

    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).

    CAS  Article  Google Scholar 

  17. 17

    Stuart, G. & Spruston, N. Determinants of voltage attenuation in neocortical pyramidal neuron dendrites. J. Neurosci. 18, 3501–3510 (1998).

    CAS  Article  Google Scholar 

  18. 18

    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).

    CAS  Article  Google Scholar 

  19. 19

    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).

    CAS  Article  Google Scholar 

  20. 20

    Williams, S.R. Spatial compartmentalization and functional impact of conductance in pyramidal neurons. Nat. Neurosci. 7, 961–967 (2004).

    CAS  Article  Google Scholar 

  21. 21

    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).

    CAS  Article  Google Scholar 

  22. 22

    Pouille, F. & Scanziani, M. Enforcement of temporal fidelity in pyramidal cells by somatic feed-forward inhibition. Science 293, 1159–1163 (2001).

    CAS  Article  Google Scholar 

  23. 23

    London, M. & Häusser, M. Dendritic computation. Annu. Rev. Neurosci. 28, 503–532 (2005).

    CAS  Article  Google Scholar 

  24. 24

    Gulledge, A.T., Kampa, B.M. & Stuart, G.J. Synaptic integration in dendritic trees. J. Neurobiol. 64, 75–90 (2005).

    CAS  Article  Google Scholar 

  25. 25

    Migliore, M. & Shepherd, G.M. Emerging rules for the distributions of active dendritic conductances. Nat. Rev. Neurosci. 3, 362–370 (2002).

    CAS  Article  Google Scholar 

  26. 26

    Häusser, M., Spruston, N. & Stuart, G.J. Diversity and dynamics of dendritic signaling. Science 290, 739–744 (2000).

    Article  Google Scholar 

  27. 27

    Magee, J.C. & Johnston, D. Plasticity of dendritic function. Curr. Opin. Neurobiol. 15, 334–342 (2005).

    CAS  Article  Google Scholar 

  28. 28

    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).

    CAS  Article  Google Scholar 

  29. 29

    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).

    CAS  Article  Google Scholar 

  30. 30

    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).

    CAS  Article  Google Scholar 

  31. 31

    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).

    CAS  Article  Google Scholar 

  32. 32

    Williams, S.R. & Stuart, G.J. Dependence of EPSP efficacy on synapse location in neocortical pyramidal neurons. Science 295, 1907–1910 (2002).

    CAS  Article  Google Scholar 

  33. 33

    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).

    Article  Google Scholar 

  34. 34

    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).

    CAS  Article  Google Scholar 

  35. 35

    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).

    CAS  Article  Google Scholar 

  36. 36

    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).

    CAS  Article  Google Scholar 

  37. 37

    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).

    CAS  Article  Google Scholar 

  38. 38

    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).

    Article  Google Scholar 

  39. 39

    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).

    CAS  Article  Google Scholar 

  40. 40

    Gulledge, A.T. & Stuart, G.J. Excitatory actions of GABA in the cortex. Neuron 37, 299–309 (2003).

    CAS  Article  Google Scholar 

  41. 43

    Stuart, G. Patch-pipet recording in brain slices. Curr. Prot. Neurosci. 6, 1–10 (1998).

    Google Scholar 

  42. 44

    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).

    CAS  Article  Google Scholar 

  43. 45

    Geiger, J.R. et al. Patch-clamp recording in brain slices with improved slicer technology. Pflugers Arch. 443, 491–501 (2002).

    CAS  Article  Google Scholar 

  44. 41

    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).

    CAS  Article  Google Scholar 

  45. 42

    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).

    CAS  Article  Google Scholar 

  46. 46

    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).

    CAS  Article  Google Scholar 

  47. 47

    Frick, A., Magee, J. & Johnston, D. LTP is accompanied by an enhanced local excitability of pyramidal neuron dendrites. Nat. Neurosci. 7, 126–135 (2004).

    CAS  Article  Google Scholar 

  48. 48

    Frick, A. & Johnston, D. Plasticity of dendritic excitability. J. Neurobiol. 64, 100–115 (2005).

    CAS  Article  Google Scholar 

  49. 49

    Dodt, H.U., Eder, M., Schierloh, A. & Zieglgansberger, W. Infrared-guided laser stimulation of neurons in brain slices. Sci. STKE, PL 2(2002).

  50. 50

    Dodt, H.U. & Zieglgansberger, W. Visualizing unstained neurons in living brain slices by infrared DIC-videomicroscopy. Brain Res. 537, 333–336 (1990).

    CAS  Article  Google Scholar 

  51. 51

    Lanni, F. & Keller, H.E. Microscopy and Microscope Optical Systems (Eds. Yuste, R., Lanni, F. & Konnerth, A.) Cold Spring Harbor Laboratory Press, 1999.

    Google Scholar 

  52. 52

    Foskett, J.K. Simultaneous Nomarski and fluorescence imaging during video microscopy of cells. Am. J. Physiol. 255, C566–C571 (1988).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

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.).

Author information

Affiliations

Authors

Corresponding author

Correspondence to Michael Häusser.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Video 1

Patch-clamp recording from a cerebellar Purkinje cell dendrite. (MOV 11261 kb)

Supplementary Video 2

Patch-clamp recording from a L5 pyramidal cell dendrite. (MOV 6067 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Davie, J., Kole, M., Letzkus, J. et al. Dendritic patch-clamp recording. Nat Protoc 1, 1235–1247 (2006). https://doi.org/10.1038/nprot.2006.164

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing