Transcriptomic Profiling Discloses Molecular and Cellular Events Related to Neuronal Differentiation in SH-SY5Y Neuroblastoma Cells

Francesco Pezzini, Laura Bettinetti, Francesca Di Leva, Marzia Bianchi, Elisa Zoratti, Rosalba Carrozzo, Filippo Maria Santorelli, Massimo Delledonne, Maciej Lalowski, Alessandro Simonati

Research output: Contribution to journalArticle

Abstract

Human SH-SY5Y neuroblastoma cells are widely utilized in in vitro studies to dissect out pathogenetic mechanisms of neurodegenerative disorders. These cells are considered as neuronal precursors and differentiate into more mature neuronal phenotypes under selected growth conditions. In this study, in order to decipher the pathways and cellular processes underlying neuroblastoma cell differentiation in vitro, we performed systematic transcriptomic (RNA-seq) and bioinformatic analysis of SH-SY5Y cells differentiated according to a two-step paradigm: retinoic acid treatment followed by enriched neurobasal medium. Categorization of 1989 differentially expressed genes (DEGs) identified in differentiated cells functionally linked them to changes in cell morphology including remodelling of plasma membrane and cytoskeleton, and neuritogenesis. Seventy-three DEGs were assigned to axonal guidance signalling pathway, and the expression of selected gene products such as neurotrophin receptors, the functionally related SLITRK6, and semaphorins, was validated by immunoblotting. Along with these findings, the differentiated cells exhibited an ability to elongate longer axonal process as assessed by the neuronal cytoskeletal markers biochemical characterization and morphometric evaluation. Recognition of molecular events occurring in differentiated SH-SY5Y cells is critical to accurately interpret the cellular responses to specific stimuli in studies on disease pathogenesis.

Original languageEnglish
Pages (from-to)1-18
Number of pages18
JournalCellular and Molecular Neurobiology
DOIs
Publication statusAccepted/In press - Jul 15 2016

Keywords

  • Axonal elongation
  • Axonal guidance signalling
  • Neuronal markers
  • RNA-seq analysis
  • Semaphorins
  • SH-SY5Y differentiation

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

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  • Cite this

    Pezzini, F., Bettinetti, L., Di Leva, F., Bianchi, M., Zoratti, E., Carrozzo, R., Santorelli, F. M., Delledonne, M., Lalowski, M., & Simonati, A. (Accepted/In press). Transcriptomic Profiling Discloses Molecular and Cellular Events Related to Neuronal Differentiation in SH-SY5Y Neuroblastoma Cells. Cellular and Molecular Neurobiology, 1-18. https://doi.org/10.1007/s10571-016-0403-y