A minimal Ksp-cadherin promoter linked to a green fluorescent protein reporter gene exhibits tissue-specific expression in the developing kidney and genitourinary tract
A minimal Ksp-cadherin promoter linked to a green fluorescent protein reporter gene exhibits tissue-specific expression in the developing kidney and genitourinary tract
复制标题
DOI：
10.1097/01.asn.0000016443.50138.cd
复制
发表时间：
2002-07-01
影响因子：
13.6
通讯作者：
Igarashi, P
中科院分区：
文献类型：
--
作者：
Shao, XL;Johnson, JE;Igarashi, P
KSP-钙粘蛋白是细胞黏附分子钙粘蛋白家族中唯一的组织特异性成员，仅在肾脏和发育中的生殖道的肾小管上皮细胞中表达。携带3425个碱基的KSP-钙粘蛋白5‘侧翼区域与LacZ报告基因相连的转基因小鼠只在肾脏表达β-半乳糖苷酶，尽管表达模式不完整(Am J Physiol277：F599-F610,1999)。为了进一步确定介导组织特异性表达的区域，制备了携带绿色荧光蛋白(GFP)报告基因5‘侧翼区1341bp或324bp的转基因小鼠。携带5‘侧翼区1341bp的转基因小鼠在所有内源性表达KSP-钙粘蛋白的胚胎组织中都表达GFP，包括输尿管芽、Wolffian管、Mullerian管以及中肾和后肾中发育中的小管。在成人肾脏中，GFP在Henle‘s环和集合管的粗大升支高表达，在近端小管和Bowman’s囊弱表达。携带324bp 5‘侧翼区的转基因小鼠仅在发育中的肾脏和胃肠道的肾小管上皮细胞中表达。免疫印迹分析表明，GFP在成年小鼠肾脏中仅有表达。综上所述，这些结果表明，KSP-钙粘蛋白5‘侧翼区的324个碱基足以在发育中的肾脏和胃肠道引导上皮特异性表达。在中肾、后肾、输尿管芽和性管中表达GFP的转基因小鼠可能对细胞谱系研究有用。
Ksp-cadherin is a unique, tissue-specific member of the cadherin family of cell adhesion molecules that is expressed exclusively in tubular epithelial cells in the kidney and developing genitourinary (GU) tract. Transgenic mice carrying 3425 bp of the Ksp-cadherin 5' flanking region linked to a lacZ reporter gene express beta-galactosidase exclusively in the kidney, although the expression pattern is incomplete (Am J Physiol 277: F599-F610, 1999). To further define the region that mediates tissue-specific expression, transgenic mice carrying 1341 bp or 324 bp of the 5' flanking region linked to a green fluorescent protein (GFP) reporter gene were produced. Transgenic mice carrying 1341 bp of the 5' flanking region expressed GFP in all embryonic tissues that endogenously express Ksp-cadherin, including the ureteric bud, Wolffian duct, Mullerian duct, and developing tubules in the mesonephros and metanephros. In the adult kidney, GFP was highly expressed in thick ascending limbs of Henle's loops and collecting ducts and weakly expressed in proximal tubules and Bowman's capsules. Transgenic mice carrying 324 bp of the 5' flanking region exhibited expression exclusively in tubular epithelia] cells in the developing kidney and GU tract. Immunoblot analysis showed that the expression of GFP was restricted to the kidney in adult mice. Taken together, these results demonstrate that 324 bp of the Ksp-cadherin 5' flanking region is sufficient to direct epithelial-specific expression in the developing kidney and GU tract. Transgenic mice that express GFP in the mesonephros, metanephros, ureteric bud, and sex ducts may be useful for cell lineage studies.