In situ recording from gut pacemaker cells

Interstitial cells of Cajal (ICC) associated with the myenteric plexus of the small intestine are crucial players in gut physiology performing pacemaker functions and directing peristalsis and segmentation. ICC have been studied after chemical isolation and under culture conditions, but concerns that these methods affect the intrinsic properties have hindered progress in our understanding of ICC. To overcome this problem, we have developed a method to obtain electrophysiological recordings from ICC in situ. The critical feature is the ability to make high resistance seals onto cells that are embedded within tissue to obtain patch clamp recordings. Our first results show a prominent presence of a chloride channel, one of the proposed ICC pacemaker channels. The developed method can be applied to auxiliary cells of the enteric nervous system such as glial cells or fibroblasts and will be ideal for the study of cell–cell communication in tissue.

[1]  G. Farrugia Interstitial cells of Cajal in health and disease , 2008, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.

[2]  S. Ward,et al.  Effects of the gap junction blocker glycyrrhetinic acid on gastrointestinal smooth muscle cells. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[3]  F. Wehner,et al.  18-β-Glycyrrhetinic Acid (BGA) as an electrical uncoupler for intracellular recordings in confluent monolayer cultures , 2001, Pflügers Archiv.

[4]  A. Nissan,et al.  Coupling among interstitial cells of Cajal in the human ileum , 2004, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.

[5]  D. Bar-Yehuda,et al.  Space-clamp problems when voltage clamping neurons expressing voltage-gated conductances. , 2008, Journal of neurophysiology.

[6]  M. Anvari,et al.  Intramuscular interstitial cells of Cajal associated with mast cells survive nitrergic nerves in achalasia , 2006, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.

[7]  Evidence for non-independent gating of P2X2 receptors expressed in Xenopus oocytes , 2002, BMC Neuroscience.

[8]  A. Reichenbach,et al.  Morphology of horseradish peroxidase (HRP)-injected glial cells in the myenteric plexus of the guinea-pig , 1994, Cell and Tissue Research.

[9]  J. Rumessen,et al.  Interstitial cells in the musculature of the gastrointestinal tract: Cajal and beyond. , 2003, International review of cytology.

[10]  J. Huizinga,et al.  Inwardly rectifying chloride channel activity in intestinal pacemaker cells. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[11]  J. Huizinga,et al.  Interstitial cells of Cajal in health and disease. Part I: Normal ICC structure and function with associated motility disorders , 2007, Histopathology.

[12]  J. Huizinga,et al.  High-conductance chloride channels generate pacemaker currents in interstitial cells of Cajal. , 2002, Gastroenterology.

[13]  K. Sanders,et al.  A Ca2+‐inhibited non‐selective cation conductance contributes to pacemaker currents in mouse interstitial cell of Cajal , 2002, The Journal of physiology.

[14]  John Malysz,et al.  W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity , 1995, Nature.

[15]  David W. Andrews,et al.  Interstitial cells of Cajal generate a rhythmic pacemaker current , 1998, Nature Medicine.

[16]  Areles Molleman,et al.  Patch Clamping: An Introductory Guide To Patch Clamp Electrophysiology , 2002 .

[17]  J. Bornstein,et al.  Intracellular recording from myenteric neurons of the guinea‐pig ileum that respond to stretch , 1998, The Journal of physiology.

[18]  S. Ward,et al.  Spontaneous electrical rhythmicity in cultured interstitial cells of Cajal from the murine small intestine , 1998, The Journal of physiology.