GEF-H1 functions in apical constriction and cell intercalations and is essential for vertebrate neural tube closure

Rho family GTPases regulate many morphogenetic processes during vertebrate development including neural tube closure. Here we report a function for GEF-H1/Lfc/ArhGEF2, a RhoA-specific guanine nucleotide exchange factor, in neurulation in Xenopus embryos. Morpholino-mediated depletion of GEF-H1 resulted in severe neural tube defects, which were rescued by GEF-H1 RNA. Lineage tracing of GEF-H1 morphants at different developmental stages revealed abnormal cell intercalation and apical constriction, suggesting that GEF-H1 regulates these cell behaviors. Molecular marker analysis documented defects in Myosin II light chain (MLC) phosphorylation, Rab11 and F-actin accumulation in GEF-H1-depleted cells. In gain-of-function studies, overexpressed GEF-H1 triggered Rho-associated kinase-dependent ectopic apical constriction, marked by apical accumulation of phospho-MLC, γ-tubulin and F-actin in superficial ectoderm, and stimulated apical protrusive activity of deep ectoderm cells. Together, our observations demonstrate novel functions of GEF-H1 in morphogenetic movements leading to neural tube closure.