ICC Clinical Science

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Figure 1.1: Overview of the distribution of ICC of Cajal in normal human sigmoid colon (panel A) and in the sigmoid colon of a patient with slow transit constipation (panel B). Each panel is made up of a montage of several fields of view. C-Kit immunoreactive structures are shown in white. A decrease in c-Kit positive immunoreactivity in all regions of the sigmoid colon is evident in panel B when compared to panel A (LM= longitudinal muscle, MY= myenteric plexus region, CM: circular muscle, SM: submucosal border). Sections cut parallel to longitudinal muscle layer for both panels. Scale bar = 100 µM for both panels.

Figure 1.2: Distribution of ICC and neuronal structures in the normal human sigmoid colon. Longitudinal muscle layer is shown in panel A, myenteric plexus region in panel B, circular smooth muscle layer in panel C, and submucosal border (yellow line with arrows) in panel D. C-Kit positive immunoreactivity is shown in red and PGP 9.5 positive immunoreactivity in gray. Scale bar = 50 µm for all panels.

Figure 1.3: Distribution of ICC and neuronal structures in human sigmoid colon taken from a patient with slow transit constipation. The longitudinal muscle layer is shown in panel A, myenteric plexus region in panel B, circular smooth muscle layer in panel C, and submucosal border (yellow line with arrows) in panel D. C-Kit positive immunoreactivity is shown in red and PGP 9.5 positive immunoreactivity in gray. Note the decreased number of immunoreactive structures for both c-Kit (ICC) and PGP 9.5 (neuronal structures). Scale bar = 50 µm for all panels.

Figure 1.4: High magnification view of ICC in the circular muscle layer. Panel A shows a single slice obtained from normal human sigmoid colon and panel B a reconstruction of 20 consecutive slices at 0.5 µm depth increments (Z axis). Note the fine processes of the individual ICC Cajal (arrow). Panels C and D show similar data from human sigmoid colon of a patient with slow transit constipation. Note the irregular surface markings and the loss of fine processes (arrow). Scale bar = 10 µM.

Figure 2.1. Computer reconstruction of c-kit immunoreactivity in cecum for both control tissue and slow transit constipation specimens. Panels represent separate reconstructions in submucosal border (SMB), circular smooth muscle layer (CM), myenteric plexus region (MPR), and longitudinal muscle layer (LM).

Figure 2.2. Computer reconstruction of c-kit immunoreactivity in ascending colon for both control tissue and slow transit constipation specimens. Panels represent separate reconstructions in submucosal border (SMB), circular smooth muscle layer (CM), myenteric plexus region (MPR), and longitudinal muscle layer (LM).

Figure 2.3. Computer reconstruction of c-kit immunoreactivity in transverse colon for both control tissue and slow transit constipation specimens. Panels represent separate reconstructions in submucosal border (SMB), circular smooth muscle layer (CM), myenteric plexus region (MPR), and longitudinal muscle layer (LM).

Figure 2.4. Computer reconstruction of c-kit immunoreactivity in sigmoid for both control tissue and slow transit constipation specimens. Panels represent separate reconstructions in submucosal border (SMB), circular smooth muscle layer (CM), myenteric plexus region (MPR), and longitudinal muscle layer (LM).

Figure 2.5. c-kit immunoreactivity expressed as percent volume for submucosal border (SMB), circular smooth muscle layer (CM), myenteric plexus region (MPR), and longitudinal muscle layer (LM). Error bars represent SEM.

Figure 3.1. Distribution of ICC. Overview of the distribution of interstitial cells of Cajal the normal human jejunum (left) and from the jejunum from the patient with insulin-dependent diabetes (right). Each panel is made up of a montage of several fields of view. C-kit immunopositive structures are shown in white. A decrease in c-kit positive immunoreactivity was seen in all regions of the jejunum in the diabetic patient compared to the control (Bar: 100 um). LM = longitudinal muscle; ICC-MY = ICC in the myenteric plexus region. CM = circular muscle.

Figure 3.2. Three-dimensional reconstruction of ICC networks. Panel A shows a three-dimensional reconstruction of the ICC-MP and panel C a three-dimensional reconstruction of ICC in the inner circular muscle layer. The panels on the left are from control tissue while the panels on the right are from tissue from the patient with diabetes. Note the decrease in c-kit immunopositivity in the myenteric plexus region of the patient compared to the control and the absence of ICC in the inner circular muscle layer of the patient. Bar: 20 um.

Figure 3.3. PGP 9.5 immunoreactivity. Nerve fiber distribution within the circular muscle layer of control tissue(left panel) and from tissue from the patient with diabetes (right panel). PGP 9.5 immunoreactivity was markedly decreased in the patient with diabetes reflecting a decrease in total number of nerve fibers within the circular muscle layer. Bar: 20 um.

Figure 3.4. Inhibitory innervation. The left hand panels show the distribution of immunopositive nerve fibers in the circular muscle layer for (A) NOS, (B) VIP, and (C) PACAP in control tissue while the right panels show the corresponding images from tissue from the diabetic patient. Immunoreactivity for nNOS, VIP, and PACAP was reduced in the patient compared to controls suggesting a decrease in inhibitory innervation. Bar: 100 um

Figure 3.5. Excitatory innervation. The left panel shows substance P immunoreactive nerve fibers in the circular muscle layer from control tissue and the right from tissue from the patient with diabetes. Substance P immunoreactivity was increased in the diabetic patient compared to controls suggesting an increase in excitatory innervation. Bar: 20 um