Reversal of axonal pathways from rhombomere 3 correlates with extra Hox expression domains

Exposure of midgastrulation mouse embryos to retinoic acid induced anteriorized expression of the Hoxa-1 (Hox-1.6) and Hoxb-1 (Hox-2.9) genes. Separate, extra domains of Hoxb-1 expression were detected as stripes and patches up to the midbrain boundary within rhombomeres r3, r2, and r1. Morphological alterations were studied in embryos of the transgenic line L17, which allowed staining of cranial ganglia, motor neurons, and axons by means of the beta-galactosidase reaction. Axons of motor neurons in r3 normally project laterally, before they turn sharply rostrally to exit with the trigeminal nerve from r2. Altered projection patterns were observed for single neurons, groups of neurons, or the complete set of r3 motor neurons in different embryos exposed to retinoic acid. Here r3 axons turned in the opposite direction and exited as facial nerves from r4. These changes of neuroectodermal fates indicate a linkage between axonal pathfinding and intrinsic neuronal specification by Hox codes.