title: Detection of 1α,25-dihydroxyvitamin D-regulated miRNAs in zebrafish by whole transcriptome sequencing.
abstract: The sterol hormone, 1α,25-dihydroxyvitamin D₃ (1α,25(OH)₂D₃), regulates gene expression and messenger RNA (mRNA) concentrations in zebrafish in vivo. Since mRNA concentrations and translation are influenced by micro-RNAs (miRNAs), we examined the influence of 1α,25(OH)₂D₃ on miRNA expression in zebrafish in vivo with whole transcriptome RNA sequencing, searched for miRNA binding sites in 1α,25(OH)₂D₃-sensitive genes, and performed correlation analyses between 1α,25(OH)₂D₃-sensitive miRNAs and mRNAs. In vehicle- and 1α,25(OH)₂D₃-treated, 7-day postfertilization larvae, between 282 and 295 known precursor miRNAs were expressed, and in vehicle- and 1α,25(OH)₂D₃-treated fish, between 83 and 122 novel miRNAs were detected. Following 1α,25(OH)₂D₃ treatment, 31 precursor miRNAs were differentially expressed (p<0.05). The differentially expressed miRNAs are predicted to potentially alter mRNAs for metabolic enzymes, transcription factors, growth factors, and Jak-STAT signaling. We verified the role of a 1α,25(OH)₂D₃-sensitive miRNA, miR125b, by demonstrating alterations in the concentrations of the mRNA of a 1α,25(OH)₂D₃-regulated gene, Cyp24a1, following transfection of renal cells with a miR125b miRNA mimic. Changes in the Cyp24a1 mRNA concentration by the miR125b miRNA mimic were associated with changes in the protein for Cyp24a1. Our data show that 1α,25(OH)₂D₃ regulates miRNA in zebrafish larvae in vivo and could thereby influence vitamin D-sensitive mRNA concentrations.
authors.affiliation: 1 Nephrology and Hypertension Research, Department of Internal Medicine, Mayo Clinic , Rochester, Minnesota.
authors.forename: Theodore A
authors.initials: TA
authors.lastname: Craig
authors.forename: Yuji
authors.initials: Y
authors.lastname: Zhang
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Magis
authors.forename: Cory C
authors.initials: CC
authors.lastname: Funk
authors.forename: Nathan D
authors.initials: ND
authors.lastname: Price
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Ekker
authors.forename: Rajiv
authors.initials: R
authors.lastname: Kumar
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017423:Sequence Analysis, RNA; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D065668:Vitamin D3 24-Hydroxylase / Q000235:genetics / Q000378:metabolism; D014815:Vitamins / Q000378:metabolism / Q000494:pharmacology; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C569474:MIRN125 microRNA, zebrafish; D035683:MicroRNAs; D014815:Vitamins; D029961:Zebrafish Proteins; C118756:dihydroxy-vitamin D3; D014807:Vitamin D; D065668:Vitamin D3 24-Hydroxylase