EURON and THEME joint PhD meeting

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86 EURON and THEME joint meeting 2011 The impact of bifunctional microRNA-9/9* on the differentiation of human ES cell – derived neural stem cells Beate Roese-Koerner, Lodovica Borghese, Laura Mürtz, Oliver Brüstle Institute for Reconstructive Neurobiology, University of Bonn. MicroRNAs are non-coding RNA molecules about 22 nucleotide-long that regulate gene expression at a post-transcriptional level. MicroRNAs are known to be involved in many cellular processes including stem cell self-renewal and differentiation. We carried out a comparative microRNA expression profiling across the differentiation of human embryonic stem cells (hESC) into neural stem cells (lt-NES®) and their neuronal progeny. MicroRNA-9/9* (miR-9/9*) was among the microRNAs that showed a strikingly differential expression pattern. It was barely detectable in hESC, whereas it was found expressed in lt-NES® and up-regulated in differentiating neurons. To gain insight into the potential roles of miR-9/9* in our long-term self-renewing lt-NES®, we made use of a lentiviral-based overexpression system and achieved overexpression of the mature forms of both miR-9 and miR-9* at comparable levels. Despite the presence of growth factors (i.e. FGF2 and EGF) in the culture medium, miR-9/9* overexpression induced a reduction in proliferation and an increase in differentiation of lt-NES®, as assessed by BrdU incorporation and expression of neuronal markers. By exposing lt-NES® to individual synthetic mimics of miR-9 and miR-9*, we could demonstrate that the promotion of differentiation is due both to miR-9 and miR-9*, whereas the anti-proliferative effect is mainly exerted by miR-9* alone. We are currently investigating potential connections between miR-9/9* and signaling pathways that are relevant in neural stem cell maintenance and differentiation. Our experimental approach provides an elegant way to assess the role of microRNAs – including bifunctional ones such as miR-9/9* - during early human neural development. It further offers a promising tool for the modulation of microRNA levels to gain control on the fate of human stem cells.
87 EURON and THEME joint meeting 2011 Retrograde tracing of neurons in the rodents lateral bladder wall: an experimental study with fluorescent latex beads: mucosal layers of the bladder vs. small intestinal mucosa Anna Schueth 1,2 , Gommert A. van Koeveringe 1 1 Maastricht University Medical Centre, azM, Department of Urology, Maastricht, The Netherlands; 2 Institute of Anatomy, University of Luebeck, Luebeck, Germany. In previous studies it was shown that latex beads (LB) of different sizes (20 – 200 nm), experimentally applied to the intact murine small intestinal mucosa, were taken up into the tissue. The LB were detected at least 20 minutes after application in different cells lying within the epithelial and subepithelial tissue of the mucosa. Both via intravital experiments by means of 2- photon microscopy (TPLSM) and confocal microscopy (CLSM) as a reference technique using dissected parts of the small intestine, the uptake and localization of LB in the small intestinal tissue could be detected and analyzed. Especially, TPLSM was used to investigate the particle uptake and further processing in anaesthetized and living mice under physiological conditions up to 8 hours. The idea is now, to transfer this technique and experimental set-up, initially described with the small intestinal mucosa, in an adapted way to mucosal layers of the bladder in rodents. With these investigations, mapping of the whole bladder and the neurons (Koebbert et al., 2000), as a matter of special interest, starting from the lateral bladder wall and ending up in the sacral cord could take place. Unlike the small intestine, the bladder has a triple innervation (pudendus and pelvic nerve, going to the sacral part and the hypogastric nerve, going to the thoracolumbar spinal cord). This could be achieved by applying the fluorescent LB to the rodent’s bladder mucosa. A subsequent tracing of the particle routes and their distribution within the lateral bladder wall could take place. Immune-histochemical techniques can be used in order to further analyse the dissected tissue.
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EURON and THEME joint PhD meeting U
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The local organizing committee: Pro
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Information about THEME 3 EURON and
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EURON a short introduction 7 EURON
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Programme 13 EURON and THEME joint
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Thursday September 22 nd 9:00 - 10:
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Friday September 23 rd 17 EURON and
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Poster presentations 19 EURON and T
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EURON Lecture Frank Kirchhoff ADDRE
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ABSTRACT 25 EURON and THEME joint m
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THEME Lecture Eckhard Mandelkow POS
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ABSTRACT 29 EURON and THEME joint m
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Abstracts 31 EURON and THEME joint
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33 Euron PhD meeting 2010 Methodolo
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Page 105 and 106: List of Participants 103 EURON and
Page 107 and 108: Nuersailike Abuduwali University of
Page 109 and 110: Marianne Eisenhardt University of B
Page 111 and 112: Sabine Klein University of Bonn PhD
Page 113 and 114: Viktoriya Peeva University of Bonn
Page 115: Sylvie van der Kruijs Maastricht Un
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EURON and THEME joint PhD meeting

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