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Visualization of the Interstitial Cells of Cajal (ICC) Network in Mice
Published on: July 27, 2011
Visualization of interstitial cells of Cajal in the mouse colon by vital staining
M Hanani1, V Louzon, S M Miller
1Laboratory of Experimental Surgery, Hadassah University Hospital, Mount Scopus, Jerusalem 91240, Israel. hananim@cc.huji.ac.il
Insights
Interstitial cells of Cajal (ICCs) are key to gut rhythm. A new DiI staining method allows better visualization of these cells in intact tissues for physiological studies.
Area of Science:
- Gastroenterology
- Cell Biology
- Neuroscience
Background:
- Interstitial cells of Cajal (ICCs) are crucial for gastrointestinal motility.
- Observing ICCs in intact tissues presents significant challenges.
Purpose of the Study:
- To develop and validate a novel staining technique for visualizing ICCs in intact gastrointestinal tissue.
- To characterize ICC morphology using this new method.
Main Methods:
- Utilized the lipophilic dye DiI to stain ICCs in freshly dissected mouse colon.
- Analyzed stained cells using light and electron microscopy.
- Quantified morphological features of identified ICC subtypes.
Main Results:
- DiI successfully labeled ICC-like cells at the submucosal-circular muscle border.
- Identified two main ICC morphologies: bipolar and multipolar cells.
- Bipolar cells exhibited longer primary processes than multipolar cells (P<0.02).
- Electron microscopy confirmed the typical ultrastructure of ICCs.
Conclusions:
- DiI staining provides a reliable method for observing ICCs in intact gastrointestinal tissues.
- This technique facilitates detailed morphological analysis and future physiological investigations of ICCs.
Abstract:
Interstitial cells of Cajal (ICCs) are believed to be a major element in generating the spontaneous rhythm of the gastrointestinal tract. A prominent problem in the study of these cells has been the difficulty in observing them in intact tissues. We used the lipophilic dye DiI to stain ICCs in the submucosal-circular muscle border of freshly dissected mouse colon. The placement of small DiI crystals in this area resulted in the labeling of ICC-like cells. Two main morphological cell types, viz., bipolar and multipolar, were noted. Bipolar cells had two primary processes emerging from the poles of an elongated soma. The mean length of these processes was 78.7 microm. These cells constituted 42.3% of the sample (n=105). Multipolar cells (54.3% of total) had a less elongated soma and extended 3-6 main processes whose mean length was 56.3 microm. These processes showed no preferred direction. The length of the primary processes of bipolar cells was 40% greater than that of multipolar cells (P<0.02). Three cells (2.9%) had only one primary process. The DiI stain could be converted into a stable electron-opaque product. Electron-microscopic observations showed that these cells had the typical appearance of ICCs reported in previous studies. This staining method should be useful for physiological investigations of ICCs in gastrointestinal tissues.
