gms | German Medical Science

61st Annual Meeting of the German Society of Neurosurgery (DGNC) as part of the Neurowoche 2010
Joint Meeting with the Brazilian Society of Neurosurgery on the 20 September 2010

German Society of Neurosurgery (DGNC)

21 - 25 September 2010, Mannheim

Transfection of ganglionic eminence-derived cells from neural precursors using proneuronal genes increases the number of DARPP-32 positive neurones

Meeting Abstract

  • Marie-Christin Pauly - Labor ofür Molekular Neurochirurgie, Abteilung für Stereotaktische Neurochirurgie, Neuorzentrum, Albert-Ludwigs-Universität Freiburg, Germany
  • Robert Kirch - Labor ofür Molekular Neurochirurgie, Abteilung für Stereotaktische Neurochirurgie, Neuorzentrum, Albert-Ludwigs-Universität Freiburg, Germany
  • Máté Döbrössy - Labor ofür Molekular Neurochirurgie, Abteilung für Stereotaktische Neurochirurgie, Neuorzentrum, Albert-Ludwigs-Universität Freiburg, Germany
  • Guido Nikkhah - Labor ofür Molekular Neurochirurgie, Abteilung für Stereotaktische Neurochirurgie, Neuorzentrum, Albert-Ludwigs-Universität Freiburg, Germany

Deutsche Gesellschaft für Neurochirurgie. 61. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC) im Rahmen der Neurowoche 2010. Mannheim, 21.-25.09.2010. Düsseldorf: German Medical Science GMS Publishing House; 2010. DocP1726

DOI: 10.3205/10dgnc197, URN: urn:nbn:de:0183-10dgnc1975

Published: September 16, 2010

© 2010 Pauly et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.


Outline

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Objective: Clinical transplantation of neuronal precursors has been shown to alter the progression of Huntington's disease (HD). Due to the limited availability of transplantable tissue, strategies to enhance the striatal population of fetal precursor cells are needed. In this study, the impact of transient transfection of two proneuronal transcription factors, Mash1 and Helt, on differentiation and maturation of ganglionic eminence-derived precursors cells is investigated.

Methods: Fetal ganglionic eminences (lateral and medial) were dissected from rat embryos on E13.5, E14.5 and E15.5. Coding regions of Mash1 and rat Helt were cloned into an expression vector under transcriptional control of elongation factor 1 alpha. Cells were grown adherent and chemically transfected for 24 hrs. Transfected cells were selected, and cultured for 7 days, and afterwards processed for immunocytochemistry.

Results: Data indicate that the expression of the transcription factors lasts about 3 to 4 days. Double transfection with Helt and Mash1 significantly increases the number of neuronal precursors (Nestin), as well as pre-mature and mature neurons (Tuj1 and Map2a/b). Furthermore, a drastic increase of DARPP-32 positive cells is observed after double transfection compared to single transfection. In addition, double transfection slightly increases the differentiation of astrocytes, but inhibits generation of oligodentrocytes. Semi-qRT-PCR reveals an increase in striatal markers on a transcriptional level 3 days after transfection already.

Conclusions: We conclude that transient transfection provides a non-hazard method for genetic manipulation of neural precursors. Transient expression of Mash1 and Helt is sufficient to increase neurogenesis and astrogenesis, and further to induce a striatal-like phenotype in vitro.