title: MicroRNA regulation of endothelin-1 mRNA in renal collecting duct cells.
abstract: OBJECTIVE
  Recently, 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.
  
  
  METHODS
  A 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.
  
  
  RESULTS
  Thirty-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.
  
  
  CONCLUSIONS
  These 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).
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Mollie E
authors.initials: ME
authors.lastname: Jacobs
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Lauren A
authors.initials: LA
authors.lastname: Jeffers
authors.affiliation: Department of Nephrology, University of Florida, Gainesville, FL 32610, USA; North Florida/South Georgia Veterans Health System, Gainesville, FL 32610, USA.
authors.forename: Amanda K
authors.initials: AK
authors.lastname: Welch
authors.affiliation: Department of Nephrology, University of Florida, Gainesville, FL 32610, USA; North Florida/South Georgia Veterans Health System, Gainesville, FL 32610, USA.
authors.forename: Charles S
authors.initials: CS
authors.lastname: Wingo
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610, USA. Electronic address: bcain@ufl.edu.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Cain
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000450:Aldosterone / Q000494:pharmacology; D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism; D001665:Binding Sites; D002460:Cell Line; D019332:Endothelin-1 / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D047468:Immunoprecipitation; D007685:Kidney Tubules, Collecting / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D020413:3' Untranslated Regions; D060565:Argonaute Proteins; D019332:Endothelin-1; D035683:MicroRNAs; D012333:RNA, Messenger; D000450:Aldosterone
keywords: Endothelin-1; Kidney; miRNA