title: Eps8 involvement in neuregulin1-ErbB4 mediated migration in the neuronal progenitor cell line ST14A.
abstract: Stable expression of the tyrosine kinase receptor ErbB4 confers increased migratory behavior to the neuronal progenitor cell line ST14A, in response to neuregulin 1 (NRG1) stimulation. We used gene expression profiling analysis to identify transcriptional changes associated with higher migratory activity caused by the activation of a specific ErbB4 isoform, and found constitutive up-regulation of the epidermal growth factor receptor pathway substrate 8 (Eps8), a multimodular regulator of actin dynamics. We confirmed the increase of Eps8, both at the mRNA and at the protein level, in stable clones expressing two different ErbB4 isoforms, both characterized by high migratory activity. Using Transwell assays and experimental manipulation of Eps8 expression level, we demonstrated that Eps8 synergizes with ErbB4 to increase both basal and ligand induced cell migration, whereas siRNA mediated Eps8 silencing strongly impairs cell motility and NRG1 induced actin cytoskeleton remodeling. By transient knockdown of Eps8 through in vivo siRNA electroporation, followed by explant primary cultures, we demonstrated that Eps8 down-regulation affects migration of normal neuronal precursors. In conclusion, our data demonstrate that Eps8 is a key regulator of motility of neuronal progenitor cells expressing ErbB4, both in basal conditions and in response to external motogenic cues.
authors.affiliation: Department of Animal and Human Biology, University of Turin, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Fregnan
authors.forename: Veselin
authors.initials: V
authors.lastname: Petrov
authors.forename: Donatella
authors.initials: D
authors.lastname: Garzotto
authors.forename: Silvia
authors.initials: S
authors.lastname: De Marchis
authors.forename: Nina
authors.initials: N
authors.lastname: Offenhäuser
authors.forename: Enrico
authors.initials: E
authors.lastname: Grosso
authors.forename: Giovanna
authors.initials: G
authors.lastname: Chiorino
authors.forename: Isabelle
authors.initials: I
authors.lastname: Perroteau
authors.forename: Giovanna
authors.initials: G
authors.lastname: Gambarotta
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D015153:Blotting, Western; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000502:physiology; D066246:ErbB Receptors / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis; D058953:Neural Stem Cells / Q000502:physiology; D020890:Neuregulin-1 / Q000378:metabolism*; D020033:Protein Isoforms / Q000502:physiology; D011506:Proteins / Q000502:physiology*; D051381:Rats; D066247:Receptor, ErbB-4; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C498142:Eps8 protein, rat; D020890:Neuregulin-1; D020033:Protein Isoforms; D011506:Proteins; C587689:ERBB4 protein, human; D066246:ErbB Receptors; C587690:Erbb4 protein, mouse; C587691:Erbb4 protein, rat; D066247:Receptor, ErbB-4