title: Vasopressin-regulated miRNAs and AQP2-targeting miRNAs in kidney collecting duct cells.
abstract: Mature microRNA (miRNA) acts as an important posttranscriptional regulator. We aimed to profile vasopressin-responsive miRNAs in kidney inner medullary collecting duct cells and to identify aquaporin-2 (AQP2)-targeting miRNAs. Microarray chip assay was carried out in inner medullary collecting duct tubule suspensions from rat kidneys in the absence or presence of desmopressin (dDAVP) stimulation (10(-9) M, 2 h). The results demonstrated 19 miRNAs, including both precursor and mature miRNAs, as potential candidates that showed significant changes in expression after dDAVP stimulation (P < 0.05). Nine mature miRNAs exhibiting >1.3-fold changes in expression on the microarray (miR-127, miR-1, miR-873, miR-16, miR-206, miR-678, miR-496, miR-298, and miR-463) were further examined by quantitative real-time PCR, and target genes of the selected miRNAs were predicted. Next, to identify AQP2-targeting miRNAs, in silico analysis was performed. Four miRNAs (miR-32, miR-137, miR-216a, and miR-216b) target the 3'-untranslated region of rat AQP2 mRNA. Target seed regions of miR-32 and miR-137 were also conserved in the 3'-untranslated region of mouse AQP2 mRNA. Quantitative real-time PCR and immunoblot analysis demonstrated that dDAVP-induced AQP2 expression was significantly attenuated in mpkCCDc14 cells when cells were transfected with miRNA mimics of miR-32 or miR-137. Moreover, luciferase reporter assay demonstrated a significant decrease of AQP2 translation in mpkCCDc14 cells transfected with miRNA mimics of miR-32 or miR-137. The present study provides novel insights into the regulation of AQP2 by RNA interference; however, vasopressin-regulated miRNAs did not include miR-32 or miR-137, indicating that the interaction of miRNAs with the AQP2 regulatory pathway requires further analysis.
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Taegu, Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University, Taegu, Korea.
authors: Jae-Eun
authors: JE
authors: Kim
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Taegu, Korea;
authors: Hyun Jun
authors: HJ
authors: Jung
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Taegu, Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University, Taegu, Korea.
authors: Yu-Jung
authors: YJ
authors: Lee
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Taegu, Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University, Taegu, Korea thkwon@knu.ac.kr.
authors: Tae-Hwan
authors: TH
authors: Kwon
mesh_terms: D000818:Animals; D051399:Aquaporin 2 / Q000378:metabolism*; D002460:Cell Line; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D007679:Kidney Medulla / Q000378:metabolism; D007685:Kidney Tubules, Collecting / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D021381:Protein Transport / Q000235:genetics; D017207:Rats, Sprague-Dawley; D014667:Vasopressins / Q000494:pharmacology*
chemical_list: D051399:Aquaporin 2; D035683:MicroRNAs; D014667:Vasopressins; D003894:Deamino Arginine Vasopressin
keywords: RNA interference; aquaporin-2; collecting duct; microRNA; vasopressin