Fig 2.

Ano1-positive ICC are decreased in all layers of the colon wall in slow transit constipation. (A) In colon tissue from control patients, Ano1 and Kit-immunolabeling showed numerous ICC in the circular muscle, submucosal border of circular muscle, myenteric plexus and the longitudinal muscle. The merged image shows overlap of Ano1 and Kit-mmunolabeling. The arrow points to a mast cell in the mucosa which was Kit-positive but negative for Ano1. (B) In colon tissue from patients with slow transit constipation, Ano1 and Kit-immunolabeling show markedly reduced ICC in the circular muscle, submucosal border of circular muscle, myenteric plexus and the longitudinal muscle as compared to the control tissue. The merged images show overlap of residual Ano1 and Kit- immunolabeling. Both panels A and B are montages of images that were collected using a 10× water immersion objective. The signal obtained with the rabbit antiserum (Ano1) is shown in red and the signal from mouse antiserum (Kit) is shown in green. (C) There were significantly fewer ICC in the circular muscle layer of the colon in patients with slow transit constipation as detected by Ano1-immunolabeling (1.89±0.13 vs. 6.43±0.37 cells per field, P <0.05, n = 4, unpaired t test). Closed symbols represent mean number of ICC in each patient per field with the standard error of mean while the open symbols represent mean number of ICC per field from all 4 patients with standard error of mean. (D) The number of ICC per field detected as Kit and Ano1-positive are identical in control patients and patients with slow transit constipation. The left part of the graph shows the number of ICC per field detected by Ano1 and Kit-immunoreactivity in colon tissue from patients with slow transit constipation (1.8 cells/field) and the right part shows similar data for colon tissue from control patients (6.4 cells/field) showing near complete overlap.