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The development of key structures within the mature vertebrate hindbrain requires the migration of neural crest (NC) cells and motor neurons to their appropriate target sites. Functional analyses in multiple species have revealed a requirement for the transcription factor gastrulation-brain-homeobox 2 (Gbx2) in NC cell migration and positioning of motor neurons in the developing hindbrain. In addition, loss of  Gbx2  function studies in mutant mouse embryos,  Gbx2 neo  , demonstrate a requirement for  Gbx2  for the development of NC-derived sensory neurons and axons constituting the mandibular branch of the trigeminal nerve (CNV). Our recent GBX2 target gene identification study identified multiple genes required for the migration and survival of NC cells (e.g.,  Robo1, Slit3, Nrp1 ). In this report, we performed loss-of-function analyses using  Gbx2 neo   mutant embryos, to improve our understanding of the molecular and genetic mechanisms regulated by  Gbx2  during anterior hindbrain and CNV development. Analysis of  Tbx20  expression in the hindbrain of  Gbx2 neo   homozygotes revealed a severely truncated rhombomere (r)2. Our data also provide evidence demonstrating a requirement for  Gbx2  in the temporal regulation of  Krox20  expression in r3. Lastly, we show that  Gbx2  is required for the expression of  Nrp1  in a subpopulation of trigeminal NC cells, and correct migration and survival of cranial NC cells that populate the trigeminal ganglion. Taken together, these findings provide additional insight into molecular and genetic mechanisms regulated by  Gbx2  that underlie NC migration, trigeminal ganglion assembly, and, more broadly, anterior hindbrain development.

Gbx2 Is Required for the Migration and Survival of a Subpopulation of Trigeminal Cranial Neural Crest Cells