title: 2-DE profiling of GDNF overexpression-related proteome changes in differentiating ST14A rat progenitor cells.
abstract: Targeted differentiation of neural progenitor cells (NPCs) is a challenge for treatment of neurodegenerative diseases by cell replacement therapy and cell signalling manipulation. Here, we applied a proteome profiling approach to the rat striatal progenitor model cell line ST14A in order to elucidate cellular differentiation processes. Native cells and cells transfected with the glial cell line-derived neurotrophic factor (GDNF) gene were investigated at the proliferative state and at seven time points up to 72 h after induction of differentiation. 2-DE combined with MALDI-MS was used to create a reference 2-DE-map of 652 spots of which 164 were identified and assigned to 155 unique proteins. For identification of protein expression changes during cell differentiation, spot patterns of triplicate gels were matched to the 2-DE-map. Besides proteins that display expression changes in native cells, we also noted 43 protein-spots that were differentially regulated by GDNF overexpression in more than four time points of the experiment. The expression patterns of putative differentiation markers such as annexin 5 (ANXA5), glucosidase II beta subunit (GLU2B), phosphatidylethanolamine-binding protein 1 (PEBP1), myosin regulatory light chain 2-A (MLRA), NASCENT polypeptide-associated complex alpha (NACA), elongation factor 2 (EF2), peroxiredoxin-1 (PRDX1) and proliferating cell nuclear antigen (PCNA) were verified by Western blotting. The results reflect the large rearrangements of the proteome during the differentiation process of NPCs and their strong modification by neurotrophic factors like GDNF.
authors.affiliation: Department of Neurology, Medical Faculty, Neurobiological Laboratory, University of Rostock, Rostock, Germany.
authors.forename: Raimund
authors.initials: R
authors.lastname: Hoffrogge
authors.forename: Susanne
authors.initials: S
authors.lastname: Beyer
authors.forename: Rayk
authors.initials: R
authors.lastname: Hübner
authors.forename: Stefan
authors.initials: S
authors.lastname: Mikkat
authors.forename: Eilhard
authors.initials: E
authors.lastname: Mix
authors.forename: Christian
authors.initials: C
authors.lastname: Scharf
authors.forename: Ulf
authors.initials: U
authors.lastname: Schmitz
authors.forename: Stefan
authors.initials: S
authors.lastname: Pauleweit
authors.forename: Matthias
authors.initials: M
authors.lastname: Berth
authors.forename: Igor Z
authors.initials: IZ
authors.lastname: Zubrzycki
authors.forename: Hilmar
authors.initials: H
authors.lastname: Christoph
authors.forename: Jens
authors.initials: J
authors.lastname: Pahnke
authors.forename: Olaf
authors.initials: O
authors.lastname: Wolkenhauer
authors.forename: Adelinde
authors.initials: A
authors.lastname: Uhrmacher
authors.forename: Uwe
authors.initials: U
authors.lastname: Völker
authors.forename: Arndt
authors.initials: A
authors.lastname: Rolfs
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D000943:Antigens, Differentiation; D051100:Glial Cell Line-Derived Neurotrophic Factor; D020543:Proteome