Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
Exp Anim. 2014;63(1):45-53. doi: 10.1538/expanim.63.45.
Serine protease inhibitor Kazal type 1 (SPINK1; mouse homologue Spink3) was initially discovered as a trypsin-specific inhibitor in the pancreas. However, previous studies have suggested that SPINK1/Spink3 is expressed in a wide range of normal tissues and tumors, although precise characterization of its gene expression has not been described in adulthood. To further analyze Spink3 expression, we generated two mouse lines in which either lacZ or Cre recombinase genes were inserted into the Spink3 locus by Cre-loxP technology. In Spink3(lacZ) mice, β-galactosidase activity was found in acinar cells of the pancreas and kidney, as well as epithelial cells of the bronchus in the lung, but not in the gastrointestinal tract or liver. Spink3(cre) knock-in mice were crossed with Rosa26 reporter (R26R) mice to monitor Spink3 promoter activity. In Spink3(cre);R26R mice, β-galactosidase activity was found in acinar cells of the pancreas, kidney, lung, and a small proportion of cells in the gastrointestinal tract and liver. These data suggest that Spink3 is widely expressed in endoderm-derived tissues, and that Spink3(cre) knock-in mice are a useful tool for establishment of a conditional knockout mice to analyze Spink3 function not only in normal tissues, but also in tumors that express SPINK1/Spink3.
丝氨酸蛋白酶抑制剂 Kazal 型 1(SPINK1;小鼠同源物 Spink3)最初在胰腺中被发现为一种胰蛋白酶特异性抑制剂。然而,先前的研究表明 SPINK1/Spink3 在广泛的正常组织和肿瘤中表达,尽管其基因表达的精确特征在成年期尚未描述。为了进一步分析 Spink3 的表达,我们利用 Cre-loxP 技术在 Spink3 基因座中插入了 lacZ 或 Cre 重组酶基因,生成了两种小鼠品系。在 Spink3(lacZ) 小鼠中,β-半乳糖苷酶活性在胰腺和肾脏的腺泡细胞以及肺部的支气管上皮细胞中被发现,但在胃肠道或肝脏中未被发现。Spink3(cre) 敲入小鼠与 Rosa26 报告基因(R26R)小鼠杂交,以监测 Spink3 启动子活性。在 Spink3(cre);R26R 小鼠中,β-半乳糖苷酶活性在胰腺、肾脏、肺部和胃肠道及肝脏的一小部分细胞中的腺泡细胞中被发现。这些数据表明 Spink3 在内胚层来源的组织中广泛表达,Spink3(cre) 敲入小鼠是建立条件性敲除小鼠以分析 Spink3 功能的有用工具,不仅在正常组织中,而且在表达 SPINK1/Spink3 的肿瘤中。