MP66-12 IDENTIFICATION OF GERM CELL SPECIFIC MIRNAS IN SEMINAL PLASMA AS EPIGENETIC BIOMARKERS PREDICTING THE QUALITY OF SPERMATOGENESIS

Abstract

You have accessJournal of UrologyInfertility: Basic Research, Physiology & Pathophysiology1 Apr 2014MP66-12 IDENTIFICATION OF GERM CELL SPECIFIC MIRNAS IN SEMINAL PLASMA AS EPIGENETIC BIOMARKERS PREDICTING THE QUALITY OF SPERMATOGENESIS Agnieszka Paradowska-Dogan, Hans-Christian Schuppe, Adrian Pilatz, Klaus Steger, and Wolfgang Weidner Agnieszka Paradowska-DoganAgnieszka Paradowska-Dogan More articles by this author , Hans-Christian SchuppeHans-Christian Schuppe More articles by this author , Adrian PilatzAdrian Pilatz More articles by this author , Klaus StegerKlaus Steger More articles by this author , and Wolfgang WeidnerWolfgang Weidner More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.2059AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Small RNA molecules, including small interfering RNAs (siRNAs), microRNAs (miRNAs), and piwi-interacting RNAs (piRNAs) have been found to be exclusively expressed in certain stages of spermatogenesis and have been detected as free circulated nucleic acids in seminal plasma of ejaculate. Vasectomy enables distinguishing between miRNAs originated form testis and epididymis, because after vasectomy seminal plasma contains only secretions of prostate and seminal vesicles. The aim of this study was to identify germ cells and Sertoli cells specific miRNA in seminal plasma as a noninvasive diagnostic parameter predicting fertility and successful sperm retrieval of infertile patients. METHODS miRNA fraction has been enriched from seminal plasma of healthy men before and 6 weeks after vasectomy as well as from patient with Sertoli Cell Only Syndrom (SCO) using HighPure miRNA Isolation Kit (Roche, USA). The expression profile of 84 miRNA-Targets from each sample applying (miRNA Finder;Qiagen) have been investigated. cDNA synthesis and preamplification was performed using universal primer mix PreAmp PCR-Mix (Qiagen) and quantitative real time PCR was carried out. The calculation of relative miRNA expression was performed using SABioscience web tool http://pcrdataanalysis.sabiosciences.com.mirna. RESULTS Differential expression of 84 known target miRNAs has been found comparing seminal plasma content prior and after vasectomy. We observed significant overexpression of 56 miRNAs (4-443 fold) 6 weeks after vasectomy, while 7 miRNAs were downregulated(0.85-2fold). The highest expression of hsa-miR-141-3p and the lowest level of hsa-miR-181a-5p has been indicated after vasectomy. Furthermore, we found 26 overexpressed miRNAs in patient with SCO (5-50 fold) and 3 miRNAs that were significantly downregulated (5-14 fold); hsa-miR-210; hsa-miR-210; hsa-miR-22-3p). The comparative analysis allowed selection of 19 testicular/epidydimal miRNAs. CONCLUSIONS In this pilot project, we successfully identified miRNAs circulating in seminal plasma and demonstrated that vasectomy modulates the expression level of 84 miRNAs. Genome wide Next Generation Sequencing and alignment of miRNAs to their target sequence will possibly unravel complex mechanism of mRNA interference for silencing of gene key factors of spermatogenesis. From clinical point of view, epigenetic biomarker in semen might facilitate diagnosis of infertility and constitute a noninvasive method predicting the outcome for surgical sperm retrieval. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e745 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Agnieszka Paradowska-Dogan More articles by this author Hans-Christian Schuppe More articles by this author Adrian Pilatz More articles by this author Klaus Steger More articles by this author Wolfgang Weidner More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...