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microRNA regulation of endothelin-1 mRNA in renal collecting duct cells

Authors
  • Jacobs, Mollie E.
  • Jeffers, Lauren A.
  • Welch, Amanda K.
  • Wingo, Charles S.
  • Cain, Brian D.1, 2, 3, 2, 4
  • 1 Department of Biochemistry and Molecular Biology
  • 2 University of Florida
  • 3 Department of Nephrology
  • 4 North Florida/South Georgia Veterans Health System
Type
Published Article
Journal
Life Sciences
Publisher
Elsevier
Publication Date
Jan 01, 2014
Accepted Date
Mar 01, 2014
Identifiers
DOI: 10.1016/j.lfs.2014.03.003
Source
Elsevier
Keywords
License
Unknown

Abstract

AimsRecently, microRNAs (miRNAs) have been implicated in control of Edn1 mRNA in several tissues. Here we examined the role of miRNA action on Edn1 mRNA expression in renal distal collecting duct cells. Main methodsA microarray study was conducted to provide a comprehensive assessment of miRNAs present in a murine inner medullary collecting duct (mIMCD-3) cell line. The experiment was designed as a comparison between mIMCD-3 cells grown in the presence and absence of aldosterone. Argonaute (Ago) immunoprecipitation experiments were used to investigate binding of the RNA induced silencing complex (RISC) to Edn1 mRNA. Key findingsThirty-four miRNAs were detected in very high abundance in mIMCD-3 cells, and a large number of others were present at lower levels. The microarray experiments were validated by quantitative PCR analysis of selected miRNAs. The microarray data, in combination with in silico examination of the Edn1 3′ UTR provided a panel of candidate miRNAs that could act upon the Edn1 expression. Edn1 mRNA was co-immunoprecipitated with an Argonaute protein antibody, and this interaction was blocked by anti-miR-709 oligonucleotides. SignificanceThese results define the miRNA landscape of the mIMCD-3 cell line. Moreover, Edn1 was shown to interact with Argonaute protein suggesting that it is a target of the RNA induced silencing complex (RISC).

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