Severe Intestinal Inflammation in the Small Intestine of Mice Induced by Controllable Deletion of Claudin-7.
Severe Intestinal Inflammation in the Small Intestine of Mice Induced by Controllable Deletion of Claudin-7.
复制标题
可控删除 Claudin-7 诱导小鼠小肠严重炎症。
DOI：
10.1007/s10620-018-4973-z
复制
发表时间：
2018-05
影响因子：
3.1
通讯作者：
Ding L
中科院分区：
文献类型：
--
作者：
Li WJ;Xu C;Wang K;Li TY;Wang XN;Yang H;Xing T;Li WX;Chen YH;Gao H;Ding L
Claudin-7 （Cldn7）作为一种潜在的肿瘤抑制基因，作为紧密连接的组成部分，可能在结直肠癌的发生发展中发挥重要作用。建立可诱导条件Cldn7在肠内的敲除小鼠模型，并分析他莫昔芬诱导后小鼠的表型。我们构建了Cldn7-flox转基因小鼠，并将其与villin-CreERT2小鼠杂交。Cldn7诱导条件敲除小鼠出生时表现正常，发育良好。我们通过给小鼠注射不同剂量的他莫昔芬诱导Cldn7基因缺失，并进一步进行表型分析。葵花籽油中他莫昔芬200μl（每次10 mg/ml）连续诱导5 d后，小鼠出现脱水、体温降低、不活动或死亡。苏木精-伊红染色结果显示，Cldn7诱导条件敲除小鼠肠道存在严重的肠道缺陷，包括上皮细胞脱落、坏死、炎症和增生。由于ICKO小鼠死亡，我们将他莫昔芬的剂量调整为每4天10 mg/ml /只（8周龄以上）的葵花籽油中100μl的剂量。我们可以诱导非典型增生和肠腺瘤。免疫荧光染色显示肠上皮结构被破坏。电镜实验分析表明，Cldn7诱导条件敲除小鼠肠内基底外膜细胞间隙增加，细胞与基质接触松动。我们建立了肠道Cldn7诱导条件敲除小鼠模型。他莫昔芬诱导小鼠肠道Cldn7缺失引发炎症和增生。
As a potential tumor suppressor gene, Claudin-7 (Cldn7), which is a component of tight junctions, may play an important role in colorectal cancer occurrence and development. To generate a knockout mouse model of inducible conditional Cldn7 in the intestine and analyze the phenotype of the mice after induction with tamoxifen. We constructed Cldn7-flox transgenic mice and crossed them with villin-CreERT2 mice. The Cldn7 inducible conditional knockout mice appeared normal and were well-developed at birth. We induced Cldn7 gene deletion by injecting different dosage of Tamoxifen into the mice and then conducted a further phenotypic analysis. After induction for five days in succession at a dose of 200μl Tamoxifen in sunflower oil at 10 mg/ml per mouse every time, the mice appeared dehydrated, had a lower temperature, and displayed inactivity or death. The results of hematoxylin-eosin staining showed that the intestines of the Cldn7 inducible conditional knockout mice had severe intestinal defects that included epithelial cell sloughing, necrosis, inflammation and hyperplasia. Owing to the death of ICKO mice, we adjusted the dose of Tamoxifen to a dose of 100μl in sunflower oil at 10 mg/ml per mouse (aged more than 8 weeks old) every four days. And we could induce atypical hyperplasia and adenoma in the intestine. Immunofluorescent staining indicated that the intestinal epithelial structure was destroyed. Electron microscopy experimental analysis indicated that the intercellular gap along the basolateral membrane of Cldn7 inducible conditional knockout mice in the intestine was increased and that contact between the cells and matrix was loosened. We generated a model of intestinal Cldn7 inducible conditional knockout mice. Intestinal Cldn7 deletion induced by tamoxifen initiated inflammation and hyperplasia in mice.
登录
查看更多内容
影响因子：
8
作者：
Bhat AA;Pope JL;Smith JJ;Ahmad R;Chen X;Washington MK;Beauchamp RD;Singh AB;Dhawan P
通讯作者：
Dhawan P
影响因子：
--
作者：
Philip R;Heiler S;Mu W;Büchler MW;Zöller M;Thuma F
通讯作者：
Thuma F
影响因子：
3.7
作者：
Okuyama T;Isoe Y;Hoki M;Suehiro Y;Yamagishi G;Naruse K;Kinoshita M;Kamei Y;Shimizu A;Kubo T;Takeuchi H
通讯作者：
Takeuchi H
影响因子：
14.9
作者：
Chan W;Costantino N;Li R;Lee SC;Su Q;Melvin D;Court DL;Liu P
通讯作者：
Liu P
影响因子：
1.5
作者：
El Marjou, F;Janssen, KP;Robine, S
通讯作者：
Robine, S