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

Heddwen L. Brooks, Shana Ageloff, Tae Hwan Kwon, William Brandt, James M. Terris, Akhil Seth, Luis Michea, Søren Nielsen, Robert Fenton, Mark A. Knepper

Research output: Contribution to journalArticlepeer-review

70 Scopus citations

Abstract

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, β-arrestin 2, neurofibromin, casein kinase IIβ, 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 aqua-porin-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 IIβ, the β-subunit of the epithelial Na channel (β-ENaC), 11β-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 IIβ, β-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.

Original languageEnglish (US)
Pages (from-to)F218-F228
JournalAmerican Journal of Physiology - Renal Physiology
Volume284
Issue number1 53-1
DOIs
StatePublished - Jan 1 2003

Keywords

  • Aldosterone
  • Epithelia
  • Kidney
  • Sodium

ASJC Scopus subject areas

  • Physiology
  • Urology

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