title: Immunoreactivity for Ano1 detects depletion of Kit-positive interstitial cells of Cajal in patients with slow transit constipation.
abstract: BACKGROUND
  Depletion of interstitial cells of Cajal (ICC) is associated with several gastrointestinal (GI) motility disorders. Changes in ICC networks are usually detected by immunolabeling for the receptor tyrosine kinase Kit. Ano1 (DOG1 or TMEM16A) was recently described as a marker of ICC in GI tract. Our aim was to determine whether Ano1 immunoreactivity can be used as a reliable marker for ICC in tissues from patients with motility disorders.
  
  
  METHODS
  Four tissues from patients with normal ICC numbers and four tissues from patients with slow transit constipation and loss of Kit-positive ICC were studied. Interstitial cells of Cajal were detected by double labeling using antisera to Kit and Ano1.
  
  
  RESULTS
  Both the processes and cell bodies of ICC in tissue from controls and slow transit constipation were immunoreactive for Ano1. There was a near complete overlap between Kit and Ano1 immunoreactivity. Tissues from patients with slow transit constipation contained significantly fewer Ano1-positive ICC than control tissues. The numbers of ICC identified by Ano1 and Kit immunoreactivity were nearly identical across the range of ICC numbers from an average of 1.64 to 7.05 cells per field and correlated with an R(2) value of 0.99.
  
  
  CONCLUSIONS
  Ano1 is a reliable and sensitive marker for detecting changes in ICC networks in humans. Labeling with antibodies selective for Ano1 reproducibly detects depletion of Kit-positive ICC in tissues from patients with slow transit constipation.
authors.affiliation: Enteric NeuroScience Program and Department of Gastroenterology and Hepatology, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
authors.forename: P
authors.initials: P
authors.lastname: Kashyap
authors.forename: P J
authors.initials: PJ
authors.lastname: Gomez-Pinilla
authors.forename: M J
authors.initials: MJ
authors.lastname: Pozo
authors.forename: R R
authors.initials: RR
authors.lastname: Cima
authors.forename: E J
authors.initials: EJ
authors.lastname: Dozois
authors.forename: D W
authors.initials: DW
authors.lastname: Larson
authors.forename: T
authors.initials: T
authors.lastname: Ordog
authors.forename: S J
authors.initials: SJ
authors.lastname: Gibbons
authors.forename: G
authors.initials: G
authors.lastname: Farrugia
mesh_terms: D000328:Adult; D000818:Animals; D000075369:Anoctamin-1; D018118:Chloride Channels; D003248:Constipation / Q000473:pathology*; D005260:Female; D005769:Gastrointestinal Motility / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D056885:Interstitial Cells of Cajal / Q000166:cytology / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000378:metabolism*; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*
chemical_list: C476213:ANO1 protein, human; D000075369:Anoctamin-1; D018118:Chloride Channels; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D019009:Proto-Oncogene Proteins c-kit