Dendritic patch-clamp recording

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.

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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.

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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.).

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Correspondence to Michael Häusser.

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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)

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