Σφακιανάκης Αλέξανδρος
ΩτοΡινοΛαρυγγολόγος
Αναπαύσεως 5 Άγιος Νικόλαος
Κρήτη 72100
00302841026182
00306932607174
alsfakia@gmail.com

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Τετάρτη 2 Δεκεμβρίου 2015

An Inducible, Large-Intestine-Specific Transgenic Mouse Model for Colitis and Colitis-Induced Colon Cancer Research

Abstract

Background

Animal models are an important tool to understand intestinal biology. Our laboratory previously generated C57BL/6-Tg(Car1-cre)5Flt transgenic mice (CAC) with large-intestine-specific Cre recombinase (Cre) expression as a model to study colon health.

Aim

To expand the utility of the CAC mouse model by determining the impact of chemically induced colitis on CAC transgene expression.

Methods

CAC mice were crossed to Rosa reporter mice (Rosa26R flox/flox ) with a lox-STOP-lox signal controlling β-galactosidase (βgal) expression and then further crossed with ApcCKO/CKO mice in some experiments to delete Apc alleles (ApcΔ580 ). Initially, 8-week-old CACTg/WT;Rosa26R flox/WT;Apc Δ580/WT mice were treated with dextran sulfate sodium (DSS) in drinking water (5 days, 0, 0.65, 1.35, or 2.0 %). Colon tissue damage and βgal labeling were analyzed 10 day after stopping DSS. Next, 8-week-old CACTg/WT;Rosa26Rflox/flox mice were treated with 0 or 1.35 % DSS, and colonic βgal labeling was assessed at 30 day post-DSS treatment. Finally, 10-week-old CACTg/WT;Apc Δ580/WT mice were treated with DSS (0 or 2 %) for 5 days and colonic tumors were analyzed at 20 weeks.

Results

CACTg/WT;Rosa26R flox/WT;Apc Δ580/WT mice had a DSS dose-dependent increase in colon epithelial damage that correlated with increased epithelial βgal labeling at 10 days (r 2 = 0.9, β = 0.75). The βgal labeling in CACTg/WT;Rosa26Rflox/flox mice colon remained high at 30 days, especially in the crypts of the healed ulcer. DSS also increased colon tumor incidence and multiplicity in CACTg/WT;Apc Δ580/WT mice.

Conclusions

DSS-mediated epithelial damage induces a persistent, Cre-mediated recombination of floxed alleles in CAC mice. This enables the examination of gene function in colon epithelium during experimental colitis and colitis-induced colon cancer.



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