ABSTRACT
Cell migration is crucial in embryonic development, tissue repair and cancer metastasis, driven by the actin and tubulin cytoskeletons that control cell shape, polarity, adhesion and movement in response to various cues. Although heterotrimeric G proteins are known to be involved in cell migration, the specific mechanisms, especially during development, remain elusive. This study examines the role of Gαi2, a heterotrimeric G-protein subunit, in cranial neural crest (NC) cell migration during Xenopus embryonic development. Our research reveals that Gαi2 interacts directly with the microtubule-associated protein EB1, regulating microtubule dynamics. We show that Gαi2 knockdown stabilizes microtubules, disrupts cell polarity and morphology, increases Rac1-GTP at the leading edge and cell-cell contacts, and impairs actin localization and focal adhesion disassembly. Additionally, RhoA-GTP is reduced at cell-cell contacts and concentrated at the leading edge in Gαi2 knockdown cells, providing evidence of a role for Gαi2 in polarity. Treatment with nocodazole, a microtubule-depolymerizing agent, reduces Rac1 activity, restoring cranial NC cell morphology, actin distribution and overall migration. Our findings highlight a crucial role for Gαi2 in cranial NC cell migration by modulating microtubule dynamics through EB1 and Rac1 activity.
Footnotes
Author contributions
Conceptualization: S.V., M.T.; Formal analysis: S.V., J.I.L., L.M.T., M.J.R., H.M.-N., R.M., M.T.; Funding acquisition: R.M., M.T.; Investigation: S.V., J.I.L., L.M.T., M.J.R., M.T.; Methodology: S.V., J.G., H.M.-N.; Project administration: M.T.; Resources: R.M., M.T.; Software: J.I.L., J.G., H.M.-N.; Supervision: M.T.; Visualization: J.I.L.; Writing – original draft: S.V., R.M., M.T.; Writing – review & editing: S.V., R.M., M.T.
Funding
This work was supported by grants from the Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT) (1180926 to M.T.), the Vicerrectoría de Investigación, Desarrollo e Innovación (VRID) (2022000483INV to M.T.), the Faculty of Biological Sciences Universidad de Concepcion (FCB-I-2022-03 to M.T.), Núcleo Milenio Información y Coordinación en Redes, ICR (NCN2024_068 to M.T.), the Medical Research Council (MR/S007792/1 to R.M.), the Biotechnology and Biological Sciences Research Council (M008517 and BB/T013044 to R.M.) and the Wellcome Trust (102489/Z/13/Z to R.M.).
Data and resource availability
All relevant data can be found within the article and its supplementary information.
