Renal principal cell-specific expression of green fluorescent protein in transgenic mice.
The purpose of this study is to develop transgenic mice with principal cell-specific expression of green fluorescent protein (GFP). After the cloning and sequencing of the mouse aquaporin-2 (AQP2) gene, 9.5 kb of the promoter were 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 analogs. GFP-expressing cells were then isolated by fluorescence-activated cell sorting. RT-PCR analysis showed expression of AQP2, AQP3, AQP4, vasopressin type 2 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 of gene expression during the development, differentiation, and maturation of renal principal cells.
Zharkikh
Ludmilla
L
Department of Pediatrics, University of Utah, Salt Lake City, Utah 84132, USA.
Zhu
Xiaohong
X
Stricklett
Peter K
PK
Kohan
Donald E
DE
Chipman
Greg
G
Breton
Sylvie
S
Brown
Dennis
D
Nelson
Raoul D
RD
D000822:Animal Husbandry; D000818:Animals; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins / Q000235:genetics; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D007679:Kidney Medulla / Q000166:cytology / Q000378:metabolism*; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice / Q000235:genetics; D008822:Mice, Transgenic / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes
C496024:Aqp2 protein, mouse; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins