Patch clamp recording from enteric neurons in situ.

The study of enteric neurons is key to understanding intestinal motility anGutn 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.