Neural stem cells (NSC) capable of differentiating into neurons, astrocytes and oligodendrocytes are a promising source of cells for the treatment of central nervous system diseases. Access to signaling proteins present in such cells in low copies and with specific regulatory functions has been very restrictive until now as judged by classical proteomic approaches and limitations due to scarcity of stem cell populations. Hence, we utilized the Kinex Antibody Microarray analysis where profiles of the proliferating porcine NSC and differentiated counterparts were compared and selected changes were verified by immunoblotting. Differentiated neural cells exhibited an increased level of RafB proto-oncogene-encoded protein-serine kinase, MAP kinase protein-serine kinase 3, heme oxygenase 2 (HO2) and protein phosphatase 4 catalytical subunit. On the other hand, relatively high level of G protein-coupled receptor-serine kinase 2 and enhanced phosphorylations of alphaB-crystallin (S45), protein-serine kinase C gamma (T655), protein-serine kinase D (PKCmu; S738+S742) together with eukaryotic translation initiation factor 2 alpha (eIF2alpha) (S51) raised intriguing questions as regards their potential functionality within stem cells. In-depth study of HO2 and phospho-S45 alphaB-crystallin confirmed expression profiles and intense cytoplasmic localization of HO2 in neurons but a weaker signal in glial cells. Phospho-S45 alphaB-crystallin was localized in nuclei of differentiated neural cells. Computer simulation of possible interaction network connecting regulated proteins, exposed additional relationships including direct interactions of HO2 with amyloid precursor protein or huntingtin-associated protein 1.

Institute of Animal Physiology and Genetics Academy of Sciences of the Czech Republic Libechov Czech Republic

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