Castrop, H. and Oppermann, M. and Weiss, Y. and Huang, Y. and Mizel, D. and Lu, H. and Germain, S. and Schweda, F. and Theilig, F. and Bachmann, S. and Briggs, J. and Kurtz, A. and Schnermann, J. (2006) Reporter gene recombination in juxtaglomerular granular and collecting duct cells by human renin promoter-Cre recombinase transgene. Physiological Genomics 25 (2), pp. 277-285.
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To assess the feasibility of using the renin promoter for expressing Cre recombinase in juxtaglomerular (JG) cells only, we generated five independent transgenic mouse lines (designated hRen-Cre) expressing Cre recombinase under control of a 12.2-kb human renin promoter. In the kidneys of adult mice Cre mRNA (RT-PCR) was found in the renal cortex, with Cre protein (immunohistochemistry) being localized in afferent arterioles and to a lower degree in interlobular arteries. Cre mRNA levels were regulated in a renin-typical fashion by changes in oral salt intake, water restriction, or isoproterenol infusion, indicating the presence of key regulatory elements within 12.2 kb of the 5'-flanking region of the human renin gene. hRen-Cre mice were interbred with both the ROSA26-EGFP and ROSA26-lacZ reporter strains to assess renin promoter activity from Cre-mediated excision of a floxed stop cassette and subsequent enhanced green fluorescent protein (EGFP) and ß-galactosidase (ß-gal) detection. In adult mice, ß-gal staining and EGFP were observed in afferent arterioles and interlobular arteries, overlapping with Cre protein expression. In addition, intense ß-gal staining was found in cortical and medullary collecting ducts where Cre expression was minimal. In embryonic kidneys, ß-gal staining was detected in the developing collecting duct system beginning at embryonic day 12, showing substantial activity of the human renin promoter in the branching ureteric bud. Our data indicate that besides its well-known activity in JG cells and renal vessels the human renin promoter is transiently active in the collecting duct system during kidney development, complicating the use of this approach for JG cell-specific excision of floxed targets.
|Institutions:|| Medicine > Lehrstuhl für Innere Medizin II|
Biology, Preclinical Medicine > Institut für Physiologie
|Keywords:||renin; kidney development|
|Subjects:||500 Science > 570 Life sciences|
600 Technology > 610 Medical sciences Medicine
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Partially|
|Deposited On:||13 Jul 2006|
|Last Modified:||05 Aug 2009 13:22|