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Visualization of interstitial cells of Cajal in living, intact tissues
Author(s) -
Hanani M.,
Belzer V.,
Rich A.,
FaussonePellegrini S.M.
Publication year - 1999
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/(sici)1097-0029(19991201)47:5<336::aid-jemt5>3.0.co;2-5
Subject(s) - interstitial cell of cajal , pathology , anatomy , myenteric plexus , staining , lucifer yellow , biology , immunohistochemistry , chemistry , microbiology and biotechnology , medicine , intracellular , gap junction
Interstitial cells of Cajal (ICC) appear to be a major element in pacing and signal transmission in the gastrointestinal tract. A prominent problem in the study of ICC has been the difficulty in observing them in intact tissues. We used several methods to visualize living ICC in freshly‐dissected tissues: (1) Placing small crystals of the lipophilic dye DiI in the submucosal‐circular muscle border in the mouse colon resulted in the labeling of living ICC‐like cells. Two main morphological cell types, bipolar and multipolar, were noted. The DiI stain could be converted into a stable, electron‐opaque product. Electron‐microscopic observations showed that the labeled cells had the typical appearance of ICC reported in previous studies. (2) Living ICC in the region of the myenteric plexus (ICC‐MP) in the small intestines of mice and guinea‐pigs were observed with Nomarski optics. This enabled the visualization of ICC in living tissues, and the impalement of the cells with Lucifer yellow‐filled microelectrodes. The dye‐labeled cells had the morphological features of ICC‐MP, and about 30% of them were found to be dye coupled to 1–21 other ICC. The identity of the cells as ICC was verified by electron‐microscopy following photoconversion, and by c‐kit immunohistochemistry. (3) Living ICC were labeled with a c‐kit antibody that does not require tissue fixation. This resulted in the fluorescent staining of the entire ICC network. Single cells were labeled by dye injection, which provided a detailed picture of ICC morphology. This method was found to be suitable for a wide range of tissues. We expect that these three methods for identifying ICC in intact, living tissues will be useful for physiological and pharmacological investigations of ICC in a variety of gastrointestinal tissues. Microsc. Res. Tech. 47:336–343, 1999. © 1999 Wiley‐Liss, Inc.