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Patch clamp recording from enteric neurons in situ

Nature Protocols volume 6, pages 15–27 (2011)Cite this article

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A Publisher Correction to this article was published on 22 October 2018

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Abstract

The study of enteric neurons is key to understanding intestinal motility and crucial to the design of therapeutic strategies for dealing with neurogenic disorders. However, enteric neurons have historically been inaccessible to patch-clamp recording. We report here the first technique that allows patch-clamp recording of neurons from the intact myenteric plexus of the mouse duodenum. The mucosa, submucosa and circular muscles are removed, exposing the myenteric plexus on the longitudinal muscle. Proteolytic treatment of exposed ganglia combined with gentle cell-surface cleaning allows gigaseal formation. Compared with previous studies using intracellular microelectrode recordings or cultured myenteric neurons, this technique provides an opportunity to explore properties of single or multiple ion channels in myenteric neurons in their native environment. The protocol—from the tissue preparation to patch-clamp recording—can be completed in ∼4 h.

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Figure 1: Equipment.
Figure 2: Schematic illustration of the perfusion system on the patch clamp setup.
Figure 3: Illustrated dissection procedure.
Figure 4: Visualization of myenteric neurons.
Figure 5: Whole-cell voltage clamp recording.
Figure 6: Whole-cell current clamp recording.

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Change history

  • 09 June 2011

    In the version of this article initially published, two important references were omitted in the discussion of the development of this method. In the fifth paragraph of the Introduction (at the top of page 16), the following sentence has been inserted before "This technique requires...": There were early demonstrations of patch clamping of myenteric neurons in the guinea pig51 and mouse52. The corresponding references are listed below: 51. Kunze, W. et al. The soma and neurites of primary afferent neurons in the guinea-pig intestine respond differently to deformation. J. Physiol. 526, 375–385 (2000). 52. Mao, Y., Wang, B. & Kunze, W. Characterization of myenteric sensory neurons in the mouse small intestine. J. Neurophysiol. 96, 998–1010 (2006). This information has been added to the HTML and PDF versions of the article.

  • 22 October 2018

    In the HTML version of this article published online, the abstract contains a typo in the first sentence: "key to understanding intestinal motility anGutn of therapeutic strategies" should read "key to understanding intestinal motility and crucial to the design of therapeutic strategies." The PDF version of the article is correct.

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Acknowledgements

This study was supported by the CNRS and by grants from the Agence Nationale de la Recherche, Fondation Schlumberger, ARCInca-2006, UPSA, IRME and Fondation pour la Recherche Médicale.

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  1. Centre de Recherche en Neurobiologie et Neurophysiologie de Marseille, Université de la Méditerranée, Marseille, France

    Nancy Osorio & Patrick Delmas

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  1. Nancy Osorio
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N.O. and P.D. conceived, designed and performed the experiments. N.O. and P.D. wrote the paper.

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Correspondence to Patrick Delmas.

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Osorio, N., Delmas, P. Patch clamp recording from enteric neurons in situ. Nat Protoc 6, 15–27 (2011). https://doi.org/10.1038/nprot.2010.172

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