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Am J Physiol Gastrointest Liver Physiol (April 16, 2009). doi:10.1152/ajpgi.00074.2009
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Submitted on February 27, 2009
Revised on April 8, 2009
Accepted on April 9, 2009

Ano1 is a selective marker of interstitial cells of Cajal in the human and mouse gastrointestinal tract

Pedro J. Gomez-Pinilla1, Simon J Gibbons1, Michael R Bardsley1, Andrea Lorincz1, Maria J. Pozo2, Pankaj J Pasricha3, Matt Van de Rijn4, Robert B. West5, Michael G Sarr6, Michael L. Kendrick, Robert R. Cima1, Eric J. Dozois1, David W. Larson1, Tamas Ordog1, and Gianrico Farrugia1*

1 Mayo Clinic
2 University of Extremadura
3 STANFORD UNIVERSITY
4 Stanford University
5 Stanford Universiity Medical Center
6 Mayo Clinic College of Medicine

* To whom correspondence should be addressed. E-mail: farrugia.gianrico{at}mayo.edu.

Populations of interstitial cells of Cajal (ICC) are altered in several gastrointestinal neuromuscular disorders. ICC are identified typically by ultrastructure and expression of Kit (CD117) a protein that is also expressed on mast cells. No other molecular marker currently exists to independently identify ICC. The expression of ANO1 (DOG1, TMEM16A), a Ca2+-activated Cl- channel, in gastrointestinal stromal tumors suggests it may be useful as an ICC marker. The aims of this study were therefore to determine the distribution of Ano1-immunoreactivity compared to Kit and to establish whether Ano1 is a reliable marker for human and mouse ICC. Expression of Ano1 in human and mouse stomach, small intestine, and colon was investigated by immunofluorescence-labeling using antibodies to Ano1 alone and in combination with antibodies to Kit. Colocalization of immunoreactivity was demonstrated by epifluorescence and confocal microscopy. In the muscularis propria, Ano1-immunoreactivity was restricted to cells with the morphology and distribution of ICC. All Ano1-positive cells in the muscularis propria were also Kit-positive. Kit-expressing mast cells were not Ano1-positive. Some non-ICC in the mucosa and submucosa of human tissues were Ano1-positive but Kit-negative. A few (3.2%) Ano1-positive cells in the human gastric muscularis propria were labeled weakly for Kit. Ano1 labels all classes of ICC and represents a highly specific marker for studying the distribution of ICC in mouse and human tissues with an advantage over Kit as it does not label mast cells.




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