A proteomic approach to studying the differentiation of neural stem cells.
The mechanisms that regulate the maintenance of stem cell self-renewal versus differentiation are complex and remain mostly unknown. Understanding neurogenesis and neural cell differentiation presents a unique challenge for the treatment of nervous system disorders. To gain more insight into molecular mechanisms of the differentiation of neural cells, we combined the advantage of porcine fetal neural stem cells (NSCs) in vitro differentiation model and proteomic analysis. Using 2-DE followed by MS, we profiled constituent proteins of NSCs and their differentiated progenies at first and then indicated protein species that were significantly up- or down-regulated during the differentiation. The largest identified group of constituent proteins was related to RNA and protein metabolism and processing, including chaperones, and the second largest consisted of proteins involved in cell organization (cytoskeleton and annexins). Differentiation of neural cells was found to be accompanied by changes in the expression of proteins involved in DNA and RNA binding, mRNA processing and transport, stress responses, iron storage, and redox regulation. Additional immunoblot analysis verified the induction of alpha-B crystallin and heterogeneous nuclear ribonucleoproteins (hnRNPs) A1 and A2/B1. Furthermore, immunocytochemistry demonstrated specific localization of alpha-B crystallin in the cytoplasm or nucleus of glial cells and confirmed cellular expression patterns of hnRNPs A1 and A2/B1. These findings represent a significant step towards understanding neural cell differentiation and identification of the regulatory proteins associated with this process.
Skalnikova
Helena
H
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov, Czech Republic.
Halada
Petr
P
Vodicka
Petr
P
Motlik
Jan
J
Rehulka
Pavel
P
Hørning
Ole
O
Chmelik
Josef
J
Nørregaard Jensen
Ole
O
Kovarova
Hana
H
D000818:Animals; D001921:Brain / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B / Q000378:metabolism; D013058:Mass Spectrometry; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D040901:Proteomics; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D013552:Swine; D038203:alpha-Crystallin B Chain / Q000378:metabolism
D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D038203:alpha-Crystallin B Chain; C439768:hnRNP A2