Src Tyrosine Kinases Regulate Neuronal Differentiation of Mouse Embryonic Stem Cells Via Modulation of Voltage-Gated Sodium Channel Activity

Kevin R. Francis, Ling Wei, Shan Ping Yu

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Voltage-gated Na+ channel activity is vital for the proper function of excitable cells and has been indicated in nervous system development. Meanwhile, the Src family of non-receptor tyrosine kinases (SFKs) has been implicated in the regulation of Na+ channel activity. The present investigation tests the hypothesis that Src family kinases influence neuronal differentiation via a chronic regulation of Na+ channel functionality. In cultured mouse embryonic stem (ES) cells undergoing neural induction and terminal neuronal differentiation, SFKs showed distinct stage-specific expression patterns during the differentiation process. ES cell-derived neuronal cells expressed multiple voltage-gated Na+ channel proteins (Nav) and underwent a gradual increase in Na+ channel activity. While acute inhibition of SFKs using the Src family inhibitor PP2 suppressed the Na+ current, chronic inhibition of SFKs during early neuronal differentiation of ES cells did not change Nav expression. However, a long-lasting block of SFK significantly altered electrophysiological properties of the Na+ channels, shown as a right shift of the current–voltage relationship of the Na+ channels, and reduced the amplitude of Na+ currents recorded in drug-free solutions. Immunocytochemical staining of differentiated cells subjected to the chronic exposure of a SFK inhibitor, or the Na+ channel blocker tetrodotoxin, showed no changes in the number of NeuN-positive cells; however, both treatments significantly hindered neurite outgrowth. These findings suggest that SFKs not only modulate the Na+ channel activation acutely, but the tonic activity of SFKs is also critical for normal development of functional Na+ channels and neuronal differentiation or maturation of ES cells.

Original languageEnglish (US)
Pages (from-to)674-687
Number of pages14
JournalNeurochemical Research
Volume40
Issue number4
DOIs
Publication statusPublished - Apr 1 2015

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Keywords

  • Neuronal differentiation
  • Sodium channels
  • Src kinases
  • Stem cells

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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