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2010 ; 138
(3
): 1068-78.e1-2
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A model to study the phenotypic changes of interstitial cells of Cajal in
gastrointestinal diseases
#MMPMID19917283
Ro S
; Park C
; Jin J
; Zheng H
; Blair PJ
; Redelman D
; Ward SM
; Yan W
; Sanders KM
Gastroenterology
2010[Mar]; 138
(3
): 1068-78.e1-2
PMID19917283
show ga
BACKGROUND & AIMS: Interstitial cells of Cajal (ICC) express the receptor
tyrosine kinase, KIT, the receptor for stem cell factor. In the gastrointestinal
(GI) tract, ICC are pacemaker cells that generate spontaneous electrical slow
waves, and mediate inputs from motor neurons. Absence or loss of ICC are
associated with GI motility disorders, including those consequent of diabetes.
Studies of ICC have been hampered by the low density of these cells and
difficulties in recognizing these cells in cell dispersions. METHODS:
Kit(+/copGFP) mice harboring a copepod super green fluorescent protein (copGFP)
complementary DNA, inserted at the Kit locus, were generated. copGFP(+) ICC from
GI muscles were analyzed using confocal microscopy and flow cytometry. copGFP(+)
ICC from the jejunum were purified by a fluorescence-activated cell sorter and
validated by cell-specific markers. Kit(+/copGFP) mice were crossbred with
diabetic Lep(+/ob) mice to generate compound Kit(+/copGFP);Lep(ob/ob) mutant
mice. copGFP(+) ICC from compound transgenic mice were analyzed by confocal
microscopy. RESULTS: copGFP in Kit(+/copGFP) mice colocalized with KIT
immunofluorescence and thus was predominantly found in ICC. In other smooth
muscles, mast cells were also labeled, but these cells were relatively rare in
the murine GI tract. copGFP(+) cells from jejunal muscles were Kit(+) and free of
contaminating cell-specific markers. Kit(+/copGFP);Lep(ob/ob) mice displayed ICC
networks that were dramatically disrupted during the development of diabetes.
CONCLUSIONS: Kit(+/copGFP) mice offer a powerful new model to study the function
and genetic regulation of ICC phenotypes. Isolation of ICC from animal models
will help determine the causes and responses of ICC to therapeutic agents.
|Animals
[MESH]
|Biomarkers/metabolism
[MESH]
|Cell Separation/methods
[MESH]
|Cells, Cultured
[MESH]
|Crosses, Genetic
[MESH]
|Diabetes Mellitus, Type 2/*metabolism/pathology
[MESH]