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Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation
Published on: March 4, 2014
The Rho guanine nucleotide exchange factor Trio is required for neural crest cell migration and interacts with
Marie-Claire Kratzer1,2, Sarah F S Becker3, Anita Grund1
1Philipps-Universität Marburg, Faculty of Biology, Molecular Embryology, 35043 Marburg, Germany.
Abstract:
Directional migration during embryogenesis and tumor progression faces the challenge that numerous external signals need to converge to precisely control cell movement. The Rho guanine exchange factor (GEF) Trio is especially well suited to relay signals, as it features distinct catalytic domains to activate Rho GTPases. Here, we show that Trio is required for Xenopus cranial neural crest (NC) cell migration and cartilage formation. Trio cell-autonomously controls protrusion formation of NC cells and Trio morphant NC cells show a blebbing phenotype. Interestingly, the Trio GEF2 domain is sufficient to rescue protrusion formation and migration of Trio morphant NC cells. We show that this domain interacts with the DEP/C-terminus of Dishevelled (DVL). DVL - but not a deletion construct lacking the DEP domain - is able to rescue protrusion formation and migration of Trio morphant NC cells. This is likely mediated by activation of Rac1, as we find that DVL rescues Rac1 activity in Trio morphant embryos. Thus, our data provide evidence for a novel signaling pathway, whereby Trio controls protrusion formation of cranial NC cells by interacting with DVL to activate Rac1.
Insights
The Rho guanine exchange factor Trio is essential for Xenopus cranial neural crest cell migration. Trio interacts with Dishevelled (DVL) to activate Rac1, controlling cell protrusion formation.
Area of Science:
- Developmental Biology
- Cell Biology
- Molecular Signaling
Background:
- Cell migration is crucial for embryogenesis and tumor progression, requiring precise control of external signals.
- Rho guanine exchange factor (GEF) Trio is a key signal relay protein with distinct catalytic domains for Rho GTPase activation.
Purpose of the Study:
- To investigate the role of Trio in Xenopus cranial neural crest (NC) cell migration and cartilage formation.
- To elucidate the molecular mechanism by which Trio regulates NC cell protrusion and migration.
Main Methods:
- Utilized Xenopus model system to study Trio's function in cranial neural crest cells.
- Employed morpholino knockdown (morphant) to assess Trio's necessity.
- Investigated the interaction between Trio's GEF2 domain and Dishevelled (DVL).
- Assessed the rescue capabilities of DVL and its domains on Trio morphant phenotypes.
- Measured Rac1 activity in Trio morphant embryos.
Main Results:
- Trio is required for Xenopus cranial neural crest cell migration and cartilage formation.
- Trio cell-autonomously controls NC cell protrusion formation; Trio morphants exhibit blebbing.
- The Trio GEF2 domain rescues protrusion formation and migration in Trio morphant NC cells.
- Trio's GEF2 domain interacts with the DEP/C-terminus of DVL.
- DVL, but not a DEP-deleted construct, rescues protrusion formation and migration in Trio morphants.
- DVL rescues Rac1 activity in Trio morphant embryos.
Conclusions:
- Trio plays a critical role in cranial neural crest cell migration and protrusion formation.
- A novel signaling pathway involves Trio interacting with DVL to activate Rac1, thereby controlling cell movement.
- This pathway is essential for cranial neural crest cell migration during embryogenesis.
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