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Am J Physiol Renal Physiol (August 30, 2001). doi:10.1152/ajprenal.0360.2000doi:10.1152/ajprenal.0360.2000
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Articles in PresS, published online ahead of print August 30, 2001
Am J Physiol Renal Physiol, 10.1152/ajprenal.0360.2000
Submitted on January 12, 2001
Accepted on July 24, 2001

Tetracycline-inducible gene expression in cultured rat rena1 collecting duct cells and in intact collecting ducts from transgenic mice

Stefania Puttini1, Ahmed T Beggah1, Antoine Ouvrard-Pascaud1, Christine Legris1, Marcel Blot-Chabaud1, Nicolette Farman1, and Frederic Jaisser1*

1 Faculte de Medicine X. Bichat, INSERM, Paris Cedex, France, France

* To whom correspondence should be addressed. E-mail: jaisser{at}bichat.inserm.fr.

The renal collecting duct (CD) plays a key role in the fine control of ion and fluid homeostasis. Several genetic diseases that involve mutations in genes encoding for ion transporters or hormone receptors specifically expressed in CD have been described. Suitable cellular or transgenic animal models expressing such mutated genes in an inducible manner should represent attractive systems for structure-function relationship analyses and the generation of appropriate physiopathological models of related diseases. Our first goal was to develop a collecting duct cell line that allows inducible gene expression using the tetracycline inducible system (tet-ON). We have designed several strategies aimed at the development of a tight and highly inducible system in RCCD1 cells, a rat cortical collecting duct (CCD) cell line exhibiting several properties of the native CCD. Analysis of reporter gene expression demonstrated that the tet-ON system is suitable for inducible gene expression in these cells. In a second step, we have tested whether transgenic tet-ON mice expressing the tet transactivator under the control of the hCMV promoter were suitable for inducible gene expression in tubule epithelial cells. The results indicated that, in vivo, the inducible expression of the LacZ reporter gene appeared to be restricted to the CD. This particular strain of transgenic mice may therefore be useful to express genes of interest in an inducible manner in the collecting duct.




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