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Articles in PresS, published online ahead of print August 6, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.0224.2001
Submitted on July 18, 2001
Accepted on July 30, 2002
1 Department of Pediatrics, University of Utah, Salt Lake City, UT, USA
2 Department of Medicine, University of Utah, Salt Lake City, UT, USA
3 Program in Membrane Biology & Renal Unit, Massachusetts General Hospital, Boston, MA, USA; Department of Medicine, Harvard Medical School, Boston, MA, USA
4 Program in Membrane Biology & Renal Unit, Massachusetts General Hospital, Boston, MA, USA
* To whom correspondence should be addressed. E-mail: Raoul.Nelson{at}hsc.utah.edu.
The purpose of this paper is to develop transgenic mice with principal cell-specific expression of green fluorescent protein (GFP). After cloning and sequencing the mouse AQP2 gene, 9.5 kb of the promoter was 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 analogues. GFP expressing cells were then isolated by fluorescence-activated cell sorting. RT-PCR analysis showed expression of AQP2, AQP3, AQP4, type 2-vasopressin 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 gene expression during the development, differentiation and maturation of renal principal cells.
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