Tetracycline-inducible gene expression in cultured rat renal CD cells and in intact CD from transgenic mice.
The renal collecting duct (CD) plays a key role in the 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 CD cell line that allows inducible gene expression using the tetracycline-inducible system (Tet-On). We 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 tetracycline transactivator under the control of the human cytomegalovirus promoter were suitable for inducible gene expression in tubule epithelial cells. The results indicate 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 for the expression of genes of interest in an inducible manner in the collecting duct.
Puttini
S
S
Institut National de la Santé et de la Recherche Médicale U478, Federative Institute of Research 02, Bichat Medical School, 75870 Paris Cedex 18, France.
Beggah
A T
AT
Ouvrard-Pascaud
A
A
Legris
C
C
Blot-Chabaud
M
M
Farman
N
N
Jaisser
F
F
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004318:Doxycycline / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005796:Genes; D017930:Genes, Reporter; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D007685:Kidney Tubules, Collecting / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*; D015533:Transcriptional Activation; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D017405:Herpes Simplex Virus Protein Vmw65; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase; D013752:Tetracycline; D004318:Doxycycline