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Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein GST-RhoAG17A from Epithelial Cell Lysates
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Trio GEF mediates RhoA activation downstream of Slit2 and coordinates telencephalic wiring
Stéphanie Backer1,2, Ludmilla Lokmane3, Camille Landragin1,2
1Institut Cochin, Université Paris Descartes, CNRS UMR 8104, 75014 Paris, France.
Development (Cambridge, England)
|September 5, 2018
Summary
Slit2 signaling activates RhoA through Trio, a guanine nucleotide exchange factor. This pathway is crucial for forebrain wiring and the proper positioning of guidepost cells and axons.
Area of Science:
- Neuroscience
- Molecular Biology
- Developmental Biology
Background:
- Trio, a Dbl family guanine nucleotide exchange factor, is known to activate Rac1 in netrin 1/DCC signaling for axon development.
- While Trio's activation of RhoA has been suggested, upstream regulators remain unidentified.
Purpose of the Study:
- To investigate the upstream factors that induce Trio-mediated RhoA activation.
- To elucidate the role of Trio in Slit2 signaling pathways.
- To examine the function of Trio in forebrain wiring and axon guidance.
Main Methods:
- Investigated Slit2-induced RhoA activation in Trio-dependent signaling.
- Utilized Trio mutant mouse embryos to assess in vivo RhoA activity.
- Analyzed the development of the ventral telencephalon and thalamocortical axons in Trio knockout mice.
Main Results:
- Demonstrated that Slit2 induces Trio-dependent RhoA activation, indicating crosstalk between Slit and Trio/RhoA signaling.
- Confirmed impaired RhoA activity in vivo within Trio mutant mouse embryos.
- Observed phenotypes in Trio knockout mice, particularly in guidepost cell positioning and thalamocortical axon pathfinding, that closely resemble those in Slit2 knockout mice.
Conclusions:
- Trio mediates RhoA activation downstream of Slit2 signaling.
- Trio plays a critical role in the precise positioning of guidepost cells and major axonal tracts.
- Established a novel function for Trio in Slit2 signaling and forebrain wiring, relevant to human brain disorders.
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