Sonic hedgehog and retinoic acid are not sufficient to induce motoneuron generation in the avian caudal neural tube.

The caudal neural tube (CNT) of the avian embryo is devoid of both dorsal and ventral roots. We show that the lack of ventral roots in the CNT, from somite 48 caudalwards, is due to an absence of post-mitotic motoneurons (MNs). The absence of MNs is not due to a defective notochordal induction since Sonic Hedgehog (SHH) signaling is intact and the caudal notochord is able to induce ectopic MNs when grafted laterally to a host neural tube. The transcription factors involved in MN specification (Pax6, Nkx6.1, and Olig2) are all expressed in the CNT, despite the lower expression level of Pax6, but an overlap between Olig2 and the ventrally expressed transcription factor Nkx2.2 is observed in the CNT. Grafting a quail CNT into the cervical level of a chick host rescues MN generation, demonstrating both the CNT potential for MN generation and the key role of the caudal environment in the MN differentiation blockade. The transplantation of the CNT-flanking somites into the cervical level does not inhibit MN generation. Furthermore, implantation of a retinoic-acid-soaked bead laterally to the CNT does not rescue MN generation. Together, these data indicate that the rostral environment contains a signal different from both SHH and Retinoic Acid that acts on MN differentiation.