Expression of an AQP2 Cre recombinase transgene in kidney and male reproductive system of transgenic mice
Expression of an AQP2 Cre recombinase transgene in kidney and male reproductive system of transgenic mice
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DOI：
10.1152/ajpcell.1998.275.1.c216
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发表时间：
1998-07-01
影响因子：
5.5
通讯作者：
Kohan, DE
中科院分区：
文献类型：
--
作者：
Nelson, RD;Stricklett, P;Kohan, DE
采用转基因小鼠的方法研究了水通道蛋白2（AQP 2）在肾集合管中主要细胞特异性表达的机制。RT-PCR和免疫细胞化学显示，小鼠AQP 2表达在肾集合管的主细胞，输精管上皮细胞和睾丸内的曲细精管。在大鼠输精管表达得到证实。RT-PCR和免疫细胞化学显示，14 kb的人5 '-侧翼区赋予核靶向和表位标记的Cre重组酶在肾集合管内的主细胞中的特异性表达，在输精管的上皮细胞中，和在转基因小鼠的睾丸内。这些结果表明，AQP 2的细胞特异性表达是在转录水平介导的，并且14 kb的人AQP 2 5 '侧翼区含有足以使AQP 2细胞特异性表达的顺式元件。最后，Cre重组酶的肾主细胞表达是实现细胞特异性基因敲除的第一步，从而允许集中检查该细胞类型中的基因功能。
A transgenic mouse approach was used to examine the mechanism of principal cell-specific expression of aquaporin-2 (AQP2) within the renal collecting duct. RT-PCR and immunocytochemistry revealed that murine AQP2 was expressed in principal cells in the renal collecting duct, epithelial cells of the vas deferens, and seminiferous tubules within testis. The vas deferens expression was confirmed in rats. RT-PCR and immunocytochemistry showed that 14 kb of the human 5'-flanking region confers specific expression of a nucleus-targeted and epitope-tagged Cre recombinase in the principal cells within the renal collecting duct, in the epithelial cells of the vas deferens, and within the testis of transgenic mice. These results suggest that cell-specific expression of AQP2 is mediated at the transcriptional level and that 14 kb of the human AQP2 5'-flanking region contain cis elements that are sufficient for cell-specific expression of AQP2. Finally, renal principal cell expression of Cre recombinase is the first step in achieving cell-specific gene knockouts, thereby allowing focused examination of gene function in this cell type.