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    MicroRNA in porcine sperms (2008)

    Art
    Poster
    Autoren
    Bergbauer, R.
    Sharbati-Tehrani, S.
    Hultschig, C.
    Einspanier, R.
    Kongress
    18. Tagung der DVG-Fachgruppe Physiologie und Biochemie
    Leipzig, 09. – 11.03.2008
    Quelle
    Abstr. in: Cermak, Rainer [Hrsg.] :Deutsche Veterinärmedizinische Gesellschaft / Fachgruppe Physiologie und Biochemie : Proceedings 18. Tagung der DVG-Fachgruppe Physiologie und Biochemie
    — S. 107
    ISBN: 978-3-934178-92-2
    Sprache
    Englisch
    Verweise
    URL (Volltext): http://www.vetmed.uni-leipzig.de/blaue-hefte/archiv/0004_DVG18-PhysiolBioch/free-online/LBH_DVG18-PhysiolBioch-Webversion.pdf
    Kontakt
    Institut für Veterinär-Biochemie

    Oertzenweg 19 b
    14163 Berlin
    Tel.+49 30 838 62225 Fax.+49 30 838-62584
    email:biochemie@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    MicroRNAs (miRNAs) are small endogenous non-coding RNA molecules and considered as major
    regulators in eukaryotic gene expression. After maturation they become single-stranded and bind to
    target mRNA molecules decreasing their translation. This process is called RNA interference (RNAi).
    MiRNAs are broadly distributed in many tissues and regulate amongst others differentiaton,
    carcinogenesis and pluripotency. There are reports of miRNAs in murine and human sperms that enter
    the oocyte at fertilisation (Amanai et al. 2006; Ostermeier et al. 2005). But the levels of these spermborne
    miRNAs are low compared to those of unfertilized oocytes, and fertilization did not alter the oocyte
    miRNA repertoire. So their role within fertilization remains unclear (Amanai et al. 2006).
    The aim of this study is to find and characterise potential regulating microRNA in porcine sperms and
    try to figure out, wether they interfere with fertilization.
    Material & Methods: Fresh porcine semen of different pig races was filtered through gaze and
    diluted with DiluPorc (Sinus Biochemistry & Electrophoresis GmbH). Swim up was used to separate the
    motile sperms from nonmotile ones and somatic cells, followed by several washing steps with PBS
    (PAA). Total RNA was isolated from agile sperms using mirVana miRNA Isolation Kit (Ambion). After
    quality control with RNA Nano Chips (Agilent Technologies) in a Agilent 2100 bioanalyser the RNA was
    labeled with fluorescent Dyes (Cy3 or Cy5, GE Healthcare) and hybridised to a miRNA microarray
    consisting of the mirVana miRNA Probe Set 1564V1Aug05 (Ambion) which was spotted onto Nexterion
    E Slides (Schott).The scanning was performed using a GenePix 4000B (Axon Instruments).
    Results: The testing of different boars revealed a set of 37 different miRNAs present in the sperms
    so far. Ten of them are present in all animals. These include miR_let_7a, let_7b, let_7c, miR_16,
    miR_191, miR_26a, miR_30d and miR_449. Five miRNAs are present in almost two animals for example
    miR_15b and miR_122a and the sperms of one animal expresses 17 miRNAs like miR_103 and
    miR_345.
    Conclusion: A compilation of different miRNAs is found in porcine sperms. Their distribution is
    obviously changing between individuals.