Effects of water restriction on gene expression in mouse renal medulla: Identification of 3βHSD4 as a collecting duct protein

Qi Cai, Maggie Keck, Matthew R. McReynolds, Janet D. Klein, Kevin Greer, Kumar Sharma, James B. Hoying, Jeff M. Sands, Heddwen L Brooks

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

To identify novel gene targets of vasopressin regulation in the renal medulla, we performed a cDNA microarray study on the inner medullary tissue of mice following a 48-h water restriction protocol. In this study, 4,625 genes of the possible ∼12,000 genes on the array were included in the analysis, and of these 157 transcripts were increased and 63 transcripts were decreased by 1.5-fold or more. Quantitative, real-time PCR measurements confirmed the increases seen for 12 selected transcripts, and the decreases were confirmed for 7 transcripts. In addition, we measured transcript abundance for many renal collecting duct proteins that were not represented on the array; aquaporin-2 (AQP2), AQP3, Pax-8, and α- and β-Na-K-ATPase subunits were all significantly increased in abundance; the β- and γ-subunits of ENaC and the vasopressin type 1A receptor were significantly decreased. To correlate changes in mRNA expression with changes in protein expression, we carried out quantitative immunoblotting. For most of the genes examined, changes in mRNA abundances were not associated with concomitant protein abundance changes; however, AQP2 transcript abundance and protein abundance did correlate. Surprisingly, aldolase B transcript abundance was increased but protein abundance was decreased following 48 h of water restriction. Several transcripts identified by microarray were novel with respect to their expression in mouse renal medullary tissues. The steroid hormone enzyme 3β-hydroxysteroid dehydrogenase 4 (3βHSD4) was identified as a novel target of vasopressin regulation, and via dual labeling immunofluorescence we colocalized the expression of this protein to AQP2-expressing collecting ducts of the kidney. These studies have identified several transcripts whose abundances are regulated in mouse inner medulla in response to an increase in endogenous vasopressin levels and could play roles in the regulation of salt and water excretion.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume291
Issue number1
DOIs
StatePublished - 2006

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3-Hydroxysteroid Dehydrogenases
Aquaporin 2
Kidney
Gene Expression
Water
Vasopressins
Proteins
Genes
Collecting Kidney Tubules
Vasopressin Receptors
Messenger RNA
Fructose-Bisphosphate Aldolase
Oligonucleotide Array Sequence Analysis
Immunoblotting
Fluorescent Antibody Technique
Real-Time Polymerase Chain Reaction
Salts
Steroids
Hormones
Enzymes

Keywords

  • Microarray
  • Vasopressin

ASJC Scopus subject areas

  • Physiology

Cite this

Effects of water restriction on gene expression in mouse renal medulla : Identification of 3βHSD4 as a collecting duct protein. / Cai, Qi; Keck, Maggie; McReynolds, Matthew R.; Klein, Janet D.; Greer, Kevin; Sharma, Kumar; Hoying, James B.; Sands, Jeff M.; Brooks, Heddwen L.

In: American Journal of Physiology - Renal Physiology, Vol. 291, No. 1, 2006.

Research output: Contribution to journalArticle

Cai, Qi ; Keck, Maggie ; McReynolds, Matthew R. ; Klein, Janet D. ; Greer, Kevin ; Sharma, Kumar ; Hoying, James B. ; Sands, Jeff M. ; Brooks, Heddwen L. / Effects of water restriction on gene expression in mouse renal medulla : Identification of 3βHSD4 as a collecting duct protein. In: American Journal of Physiology - Renal Physiology. 2006 ; Vol. 291, No. 1.
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