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Articles in PresS, published online ahead of print September 11, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00054.2002
Submitted on February 7, 2002
Accepted on September 3, 2002
1 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, Bethesda, MD, USA
2 Department of Cell Biology, Institute of Anatomy, University of Aarhus, Aarhus, Jutland, Denmark; Water and Salt Institute, Aarhus University, Aarhus, Jutland, Denmark
3 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, Bethesda, MD, USA; Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, Bethesda, MD, USA
4 Department of Cell Biology, Institute of Anatomy, University of Aarhus, Aarhus, Jutland, Denmark
* To whom correspondence should be addressed. E-mail: knep{at}helix.nih.gov.
With the aim of identifying possible gene targets for direct or indirect regulation by vasopressin in the renal medulla, we have carried out cDNA array experiments in inner medullae of Brattleboro rats infused with the V2 receptor-selective vasopressin analog dDAVP for 72 hours. Of the 1176 genes on the array, 137 transcripts were increased by 2 fold or more, and 10 transcripts were decreased to 0.5 fold or less. Quantitative, real-time RT-PCR measurements confirmed increases seen for 6 selected transcripts (WT1, ß-arrestin 2, neurofibromin, casein kinase II , aquaporin-3, and aquaporin-4). To correlate changes in mRNA expression with changes in protein expression, we carried out quantitative immunoblotting for 28 of the proteins whose cDNAs were on the array. For several targets including aquaporin-2, transcript abundance and protein abundance changes did not correlate. However, for most genes examined, changes in mRNA abundances were associated with concomitant protein abundance changes. Targets with demonstrated increases in both protein and mRNA abundances included neurofibromin, casein kinase IIß, ß-ENaC, 11-ß hydroxysteroid dehydrogenase type 2 (11ßHSD2), and c-Fos. Additional cDNA arrays revealed that several transcripts that were increased in abundance after 72 hours of dDAVP were also increased after 4 hours, including casein kinase IIß, ß-ENaC, aquaporin-3, UT-A, and syntaxin 2. These studies have identified several transcripts whose abundances are regulated in the inner medulla in response to infusion of dDAVP, and which could play roles in regulation of salt and water excretion.
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