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Inducible Podocyte-Specific Gene Expression in Transgenic Mice

Tetsuya Shigehara^*, Concepcion Zaragoza^*, Chagriya Kitiyakara^*, Hideko Takahashi^*, Huiyan Lu^*, Marcus Moeller, Lawrence B. Holzman and Jeffrey B. Kopp^*

*Kidney Disease Section, Metabolic Diseases Branch, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland; and Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan.

Correspondence to Dr. Jeffrey Kopp, 10 Center Drive, MSC 1268, Bethesda, MD 20892. Phone: 301-594-3403; Fax: 301-402-0014;

ABSTRACT. The podocyte plays a key role in glomerular function and glomerular disease. To facilitate studies of podocyte function, we have developed a transgenic mouse model with inducible expression in the podocyte. The tetracycline-inducible transgenic system facilitates gene expression with restricted cellular distribution and tight temporal control. Recently, Bujard and colleagues have developed a functionally improved reverse tetracycline–controlled transcriptional activator (rtTA) with substantially lower background in the off state (the absence of tetracycline) and greater inducibility in the on state (the presence of tetracycline). We used the human podocin (NPHS2) gene promoter to control expression of the rtTA cassette and bred these mice with a reporter mouse line that contains the cytomegalovirus minimal promoter and tetO promoter elements together with LacZ, encoding -galactosidase. Dual transgenic mice, bearing both podocin-rtTA and tetO-LacZ transgenes, had no detectable expression in kidney or other organs in the absence of tetracycline. Administration of tetracycline in the drinking water was associated with podocyte expression of -galactosidase, in a fashion that was time dependent (maximal at 1 wk) and dose-dependent (maximal at 2 mg/ml). Podocyte expression was confirmed in two ways: histochemical staining for -galactosidase and double-immunostaining using the podocyte marker WT-1 and -galactosidase. This transgenic system should aid future investigations of podocyte function. E-mail: jbkopp@nih.gov

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Copyright © 2008 by the American Society of Nephrology. Online ISSN: 1533-3450 Print ISSN: 1046-6673