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1 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892; and 2 Department of Cell Biology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark
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 medullas of Brattleboro rats infused with the V2
receptor-selective vasopressin analog desamino-Cys1,D-Arg8
vasopressin (dDAVP) for 72 h. Of the 1,176 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 six selected transcripts
(Wilms' tumor protein, 
-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 II, the -subunit of the epithelial Na channel
(-ENaC), 11-hydroxysteroid dehydrogenase type 2, and c-Fos.
Additional cDNA arrays revealed that several transcripts that were
increased in abundance after 72 h of dDAVP were also increased
after 4 h, including casein kinase II, -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 that could play roles in the regulation of salt
and water excretion.
aldosterone; kidney; sodium; epithelia
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