A Src-Tks5 Pathway Is Required for Neural Crest Cell Migration during Embryonic Development

Abstract

In the adult organism, cell migration is required for physiological processes such as angiogenesis and immune surveillance, as well as pathological events such as tumor metastasis. The adaptor protein and Src substrate Tks5 is necessary for cancer cell migration through extracellular matrix in vitro and tumorigenicity in vivo. However, a role for Tks5 during embryonic development, where cell migration is essential, has not been examined. We used morpholinos to reduce Tks5 expression in zebrafish embryos, and observed developmental defects, most prominently in neural crest-derived tissues such as craniofacial structures and pigmentation. The Tks5 morphant phenotype was rescued by expression of mammalian Tks5, but not by a variant of Tks5 in which the Src phosphorylation sites have been mutated. We further evaluated the role of Tks5 in neural crest cells and neural crest-derived tissues and found that loss of Tks5 impaired their ventral migration. Inhibition of Src family kinases also led to abnormal ventral patterning of neural crest cells and their derivatives. We confirmed that these effects were likely to be cell autonomous by shRNA-mediated knockdown of Tks5 in a murine neural crest stem cell line. Tks5 was required for neural crest cell migration in vitro, and both Src and Tks5 were required for the formation of actin-rich structures with similarity to podosomes. Additionally, we observed that neural crest cells formed Src-Tks5-dependent cell protrusions in 3-D culture conditions and in vivo. These results reveal an important and novel role for the Src-Tks5 pathway in neural crest cell migration during embryonic development. Furthermore, our data suggests that this pathway regulates neural crest cell migration through the generation of actin-rich pro-migratory structures, implying that similar mechanisms are used to control cell migration during embryogenesis and cancer metastasis.

Prognostic significance of Tks5 expression in gliomas

“…18,22 Moreover, several proteins involved in actin and membrane regulation that are critical for invadopodia functional activity bind to Tks5 SH3 domains and/or Src phosphorylation sites (for example, tyrosine Y557) including N-WASP, dynamin and Nck adaptor proteins. [22][23][24][25] Recently, an essential role for a Src-Tks5 signalling pathway involving Tks5 tyrosine phosphorylation was uncovered in migration of neural crest-derived cells during zebrafish embryogenesis 26 which supports the notion that developmental cell migrations may provide default mechanisms that drive invasion and metastasis. 26 The transcription factor Twist1, which can stimulate the epithelial-mesenchymal transition, induces invadopodia formation and metastasis via a Src-Tks5 dependent pathway.…”

“…[22][23][24][25] Recently, an essential role for a Src-Tks5 signalling pathway involving Tks5 tyrosine phosphorylation was uncovered in migration of neural crest-derived cells during zebrafish embryogenesis 26 which supports the notion that developmental cell migrations may provide default mechanisms that drive invasion and metastasis. 26 The transcription factor Twist1, which can stimulate the epithelial-mesenchymal transition, induces invadopodia formation and metastasis via a Src-Tks5 dependent pathway. 27 The actin filament-binding protein cortactin, which additionally promotes actin polymerization and branching by activating the Arp2/3 actin nucleation complex, is also a central mediator of invadopodia formation.…”

Sulforaphane protects against ethanol‐induced oxidative stress and apoptosis in neural crest cells by the induction of Nrf2‐mediated antioxidant response

“…To determine whether SFN can reduce ethanol-induced cell death in NCCs, JoMa 1.3 cells were treated with ethanol or/and SFN at different concentrations for 24 h or treated with 100 mM ethanol or/and 1 mM SFN for 24, 48 or 72 h. JoMa 1.3 cells were chosen as a model for the proposed studies because (i) they are NCCs derived from mouse embryos; (ii) this cell line expresses early NCC markers and can be instructed to differentiate into neurons, glia, smooth muscle cells, melanocyte and chondrocytes (Maurer et al, 2007); and (iii) this cell line represents a powerful tool for studying the mechanisms of NCC development (Murphy et al, 2011) and has been used to elucidate the role of miRNAs in NCC development (Cordes et al, 2009;Sheehy et al, 2010). As shown in Figure 1A-C, while SFN at 0.5, 1 or 2 mM and at 0.25, 0.5, 1 or 2 mM significantly reduced cell death in NCCs exposed to 50 and 100 mM ethanol, respectively, SFN at the concentrations up to 4 mM did not significantly diminish cell death in NCCs treated with 200 mM ethanol.…”

Frank-ter Haar Syndrome Protein Tks4 Regulates Epidermal Growth Factor-dependent Cell Migration

“…In this process Nck adaptor proteins, Nck1 and Nck2, seem to link Tks5 to invadopodia actin regulation and extracellular matrix degradation (10). Finally, Tks5 has been shown to be required for migration of neural crest cell during development of zebrafish embryos (11).…”