cDNA array identification of genes regulated in rat renal medulla in response to vasopressin infusion

• Published in: American journal of physiology. Renal physiology 2003-01, Vol.284 (1), p.F218-F228
• Main Authors: Brooks, Heddwen L, Ageloff, Shana, Kwon, Tae-Hwan, Brandt, William, Terris, James M, Seth, Akhil, Michea, Luis, Nielsen, Soren, Fenton, Robert, Knepper, Mark A
• Format: Article
• Language: English
• Subjects: 11-beta-Hydroxysteroid Dehydrogenase Type 2; Aldosterone - physiology; Animals; Deamino Arginine Vasopressin - pharmacology; Gene Expression - drug effects; Hydroxysteroid Dehydrogenases - genetics; Kidney Medulla - drug effects; Kidney Medulla - physiology; Male; Oligonucleotide Array Sequence Analysis - standards; Rats; Rats, Brattleboro; Renal Agents - pharmacology; Reproducibility of Results; RNA, Messenger - analysis; Sodium - metabolism; Sodium-Potassium-Exchanging ATPase - genetics; Vasopressins - genetics
• ISSN: 1931-857X; 1522-1466
• DOI: 10.1152/ajprenal.00054.2002

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 V(2) 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, beta-arrestin 2, neurofibromin, casein kinase IIbeta, 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 IIbeta, the beta-subunit of the epithelial Na channel (beta-ENaC), 11beta-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 IIbeta, beta-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.