Renal principal cell-specific expression of green fluorescent protein in transgenic mice.
Renal principal cell-specific expression of green fluorescent protein in transgenic mice.
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DOI：
10.1152/ajprenal.0224.2001
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发表时间：
2002-12
期刊：
影响因子：
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通讯作者：
L. Zharkikh;Xiaohong Zhu;P. Stricklett;D. Kohan;Greg Chipman;S. Breton;Dennis Brown;R. Nelson
中科院分区：
文献类型：
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作者：
L. Zharkikh;Xiaohong Zhu;P. Stricklett;D. Kohan;Greg Chipman;S. Breton;Dennis Brown;R. Nelson
本研究的目的是建立绿色荧光蛋白（GFP）主要细胞特异性表达的转基因小鼠。在克隆和测序小鼠水通道蛋白2（AQP 2）基因后，使用9.5kb的启动子驱动GFP在转基因小鼠中表达。在转基因小鼠中，GFP选择性地表达在肾集合管的主细胞中，而不是在闰细胞中。小鼠脱水后表达增加。AQP 2和GFP的表达维持在原代培养的肾髓质，刺激cAMP或加压素类似物。然后通过荧光激活细胞分选分离GFP表达细胞。RT-PCR分析显示AQP 2、AQP 3、AQP 4、加压素2型受体和cAMP反应元件结合蛋白表达，但不表达H+-ATP酶B1亚基或阴离子交换蛋白1。在培养中扩增这些细胞后，RT-PCR分析显示相同基因的持续表达。这种基因表达模式是主细胞而不是闰细胞的模式。该转基因小鼠模型可用于肾主细胞发育、分化和成熟过程中基因表达的进一步研究。
The purpose of this study is to develop transgenic mice with principal cell-specific expression of green fluorescent protein (GFP). After the cloning and sequencing of the mouse aquaporin-2 (AQP2) gene, 9.5 kb of the promoter were used to drive expression of GFP in transgenic mice. In transgenic mice, GFP was selectively expressed in principal cells of the renal collecting duct and not in intercalated cells. Expression was increased by dehydration of mice. AQP2 and GFP expression was maintained in primary cultures of renal medulla that were stimulated with cAMP or vasopressin analogs. GFP-expressing cells were then isolated by fluorescence-activated cell sorting. RT-PCR analysis showed expression of AQP2, AQP3, AQP4, vasopressin type 2 receptor, and cAMP response element binding protein but not H+-ATPase B1 subunit or anion exchanger 1. After expansion of these cells in culture, RT-PCR analysis showed continued expression of the same genes. This pattern of gene expression is that of principal cells rather than intercalated cells. This transgenic mouse model can be used in future studies of gene expression during the development, differentiation, and maturation of renal principal cells.