Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or . mil. Before sharing sensitive information, make sure you’re on a federal government site. VSports app下载.

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. V体育官网.

Comparative Study
. 2008 Sep;14(9):979-84.
doi: 10.1038/nm.1865.

An efficient and versatile system for acute and chronic modulation of renal tubular function in transgenic mice

Milena Traykova-Brauch  1 , Kai Schönig, Oliver Greiner, Tewfik Miloud, Anna Jauch, Manja Bode, Dean W Felsher, Adam B Glick, David J Kwiatkowski, Hermann Bujard, Jürgen Horst, Magnus von Knebel Doeberitz, Felix K Niggli, Wilhelm Kriz, Hermann-Josef Gröne, Robert Koesters
Affiliations
Comparative Study

An efficient and versatile system for acute and chronic modulation of renal tubular function in transgenic mice

Milena Traykova-Brauch et al. Nat Med. 2008 Sep.

Abstract

We describe a transgenic mouse line, Pax8-rtTA, which, under control of the mouse Pax8 promoter, directs high levels of expression of the reverse tetracycline-dependent transactivator (rtTA) to all proximal and distal tubules and the entire collecting duct system of both embryonic and adult kidneys. Using crosses of Pax8-rtTA mice with tetracycline-responsive c-MYC mice, we established a new, inducible model of polycystic kidney disease that can mimic adult onset and that shows progression to renal malignant disease. When targeting the expression of transforming growth factor beta-1 to the kidney, we avoided early lethality by discontinuous treatment and successfully established an inducible model of renal fibrosis. Finally, a conditional knockout of the gene encoding tuberous sclerosis complex-1 was achieved, which resulted in the early outgrowth of giant polycystic kidneys reminiscent of autosomal recessive polycystic kidney disease. These experiments establish Pax8-rtTA mice as a powerful tool for modeling renal diseases in transgenic mice VSports手机版. .

PubMed Disclaimer

Figures

Figure 1
Figure 1
Pax8-rtTA–mediated, doxycycline-controlled β-galactosidase expression in adult tissues. (a) The Pax8 promoter directs the expression of rtTA, which, in the presence of doxycycline (Dox), binds and transactivates the tetracycline-responsive element (TRE). The TRE is a bidirectional Ptet and allows the tetracycline-controlled expression of two target genes at a time. In the case of NZL-2 mice, these are two reporter genes encoding luciferase and nuclear β-galactosidase. (b) Enzymatic X-gal staining of cryosections of kidneys derived from Pax8-rtTA/NZL-2 mice either uninduced (without Dox) or induced with doxycycline over a period of 10 d (+Dox). β-galactosidase positivity was strictly dependent on the prior administration of Dox, and it was exclusively found in the tubular epithelium of the kidney, leaving glomeruli and blood vessels unstained. Some periportally located hepatocytes in the liver also stained positively; all other major organs, including the thyroid gland, were negative.
Figure 2
Figure 2
Pax8-rtTA–mediated, doxycycline-controlled β-galactosidase expression in adult renal tubular cells. (a) β-galactosidase positivity (nuclear staining) in Pax8-rtTA/NZL-2 double-transgenic mice induced with doxycycline was found in all renal tubular epithelial cells, including cortical and medullary regions. (b) Parietal and visceral epithelial cells, as well as mesangial and endothelial cells, of the glomeruli were unstained. Only proximal tubular epithelial cells in a parietal position stained positively in Pax8-rtTA/NZL-2 mice. (c,d) A virtually identical staining pattern to that in b in Pax8-rtTA/LC-1/Rosa26R triple-transgenic mice induced with doxycycline. This experiment uses a transgene encoding tetracycline-inducible Cre recombinase (LC-1) and a gene encoding a cytoplasmic β-galactosidase reporter (Rosa26R). Co, cortex; os, outer stripe of medulla; is, inner stripe of medulla; g, glomerulus; pt, proximal tubular cells in parietal position.
Figure 3
Figure 3
Pax8-rtTA–mediated, doxycycline-controlled β-galactosidase expression in embryonic kidney. (a) Whole-mount embryo (E10.5) stained for β-galactosidase activity. β-galactosidase–positive cells were macroscopically visible in three locations: mid-hindbrain region (mh), hindbrain (hi) and cloaca (cl). (b) Cross-section through whole-mount embryo (E10.5) at abdominal level stained for β-galactosidase activity. β-galactosidase–positive cells were found in mesonephric tubules (mt) and Wolffian duct (wd). (c) Cross-section through whole-mount embryo (E10.5) at a more caudal level than in b. β-galactosidase–positive cells were found in Wolffian duct and the ureteric bud (ub) invading the metanephric mesenchyme. Rp, prospective renal pelvis. (d) Whole-mount embryonic kidney (E17.5) stained for β-galactosidase activity. β-galactosidase positivity was strictly dependent on the prior administration of doxycycline. (e) Longitudinal cross-section of induced embryonic kidney (E17.5) stained for β-galactosidase activity. Expression was found in maturing tubular epithelium and developing collecting ducts (cd). No β-galactosidase activity was detected in the early nephron anlagen or in developing or mature glomeruli. Om, outer medulla; im, inner medulla.
Figure 4
Figure 4
Time course of transgene induction in Pax8-rtTA/LC-1–transgenic mice upon doxycycline administration. (a) Time course of luciferase induction in three individual double-transgenic mice (exposure time, 180 s). Signal intensities are plotted relative to the time after doxycycline treatment. (b) Whole-body bioluminescence of kidney-specific luciferase expression in a representative double-transgenic mouse before and after 6 h of treatment with doxycycline. The relative luminescence intensity is indicated by a color scale bar. (c) Cycle times of luciferase gene activation and deactivation via doxycycline was monitored for 28 d in Pax8-rtTA/LC-1–transgenic mice (exposure time, 1 s). The signal intensities from four mice per group were averaged and plotted relative to the time. Mice were treated with doxycycline as follows: group 1, at days 1–28; group 2, at days 1–5 and 25–28; group 3, at days 1 and 2 and 25 and 26. On day 25, treatment with doxycycline was started just after imaging. (d) One representative mouse for each of the three experimental groups in c is presented (signal intensity scale bar as in b). Data represent means ± s.d.
Figure 5
Figure 5
Renal disease models. (a–d) Histological analysis of polycystic kidneys showing different stages of malignant progression induced in Pax8-rtTA/tetO-MYC double-transgenic mice. Doxycycline was continuously administered to 3-month-old Pax8-rtTA/tetO-MYC double-transgenic mice over a period of 4 months. (a) Polycystic kidney with renal adenoma and renal cell carcinoma (rcc). (b) Renal adenoma (a) magnified from marked box in a. (c) Polycystic kidney with tubular (tc) and glomerular (gc) cysts and renal cell carcinoma. (d) Renal cell carcinoma cells magnified from c. (e,f) Renal fibrosis induced in Pax8-rtTA/tetO–Tgf-β1 double-transgenic mice. Pax8-rtTA/tetO–Tgf-β1 double-transgenic mice (3 months old) were treated for six consecutive cycles (2 d with doxycycline, 5 d without doxycycline). Overexpression of Tgf-β1 led to massive extracellular matrix accumulation and prominent renal fibrosis, as indicated here by collagen I immunohistochemistry in noninduced (e) or induced (f) mice. (g,h) Severe PKD induced in Pax8-rtTA–mediated, tetracycline-induced Tsc1-knockout mice. Pregnant Pax8-rtTA/LC-1/Tsc1flox/flox mice were treated with doxycycline from the first day of mating until delivery. After birth, doxycycline treatment was stopped. Giant polycystic kidneys developed in 3-week-old Pax8-rtTA/LC-1/Tsc1flox/flox offspring mice as shown in g. PAS staining in h revealed extensive cyst formation in all tubular segments and occasional protein deposits (p) caused by proteinuria.
See this image and copyright information in PMC

VSports最新版本 - References

    1. Gossen M, Bujard H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA. 1992;89:5547–5551. - PMC - PubMed
    1. Gossen M, et al. Transcriptional activation by tetracyclines in mammalian cells. Science. 1995;268:1766–1769. - PubMed
    1. Kistner A, et al. Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. Proc. Natl. Acad. Sci. USA. 1996;93:10933–10938. - V体育安卓版 - PMC - PubMed
    1. Poleev A, et al. PAX8, a human paired box gene: isolation and expression in developing thyroid, kidney and Wilms’ tumors. Development. 1992;116:611–623. - VSports注册入口 - PubMed
    1. Plachov D, et al. Pax8, a murine paired box gene expressed in the developing excretory system and thyroid gland. Development. 1990;110:643–651. - PubMed (V体育官网入口)

Publication types

MeSH terms

Substances