The vertebrate neural crest arises at the border of the neural plate during early stages of nervous system development; however, little is known about the molecular mechanisms underlying neural crest formation. Here we identify a secreted protein, Noelin-1, which has the ability to prolong neural crest production. Noelin-1 messenger RNA is expressed in a graded pattern in the closing neural tube. It subsequently becomes restricted to the dorsal neural folds and migrating neural crest. Over expression of Noelin-1 using recombinant retroviruses causes an excess of neural crest emigration and extends the time that the neural tube is competent to generate as well as regenerate neural crest cells. These results support an important role for Noelin-1 in regulating the production of neural crest cells by the neural tube.

Early stages in chick neurogenesis were investigated with tritiated thymidine (3H-Tdr) autoradioraphy to determine the location and identity of the first neurons produced for the central nervous system. These cells have been shown to arise prior to neural tube closure (Sechrist, '75). Chicks were treated at selected intervals between 20 and 72 hours of incubation with 3H-Tdr in a modified pulse-labeling technique, and terminated on the 18th day of embryonic development (E18), when neuronal types could be determined. Some of the earliest neurons start their final DNA synthesis before 20 hours of incubation (head process, Hamburger-Hamilton stage 5). These are primarily medium-sized cells of the reticular formation in the medulla and at the diencephalic-mesencephalic junction, but also in the intermediate zone of the spinal cord. Motor neurons of the brainstem and spinal cord begin to appear next, after 26-28 hours incubation; the first sensory neurons arise after 32 hours. Other workers (Ramon y Cajal, '60; Tello, '23; Windle and Austin, '36) found that neurons of the reticular formation were the first to differentiate neurofibrils, during the latter part of E2, indicating that fibrillogenesis in these cells begin about 24 hours after the initial cessation of DNA replication.

The largest of the cranial ganglia, the trigeminal ganglion relays cutaneous sensations of the head to the central nervous system. Its sensory neurons have a dual origin from both ectodermal placodes and neural crest. Here, we show that birth of neurons derived from the chick ophthalmic trigeminal placode begins prior to their ingression (HH11), as early as HH8, and considerably earlier than previously suspected (HH16). Furthermore, cells exiting the cell cycle shortly thereafter express the ophthalmic trigeminal placode marker Pax3 (HH9). At HH11, these post-mitotic Pax3+ placode cells begin to express the pan neuronal marker, neurofilament, while still in the ectoderm. Analysis of the ectodermal origin and distribution of these early post-mitotic neurons reveals that the ophthalmic placode extends further rostrally than anticipated, contributing to neurons that reside in and make a significant contribution to the ophthalmic trigeminal nerve. These data redefine the timing and extent of neuron formation from the ophthalmic trigeminal placode.

Previous studies have suggested that the rostrocaudal patterning of branchial arches in the vertebrate embryo derives from a coordinate segmental specification of gene expression in rhombomeres (r) and neural crest. However, expression of the Krox‐20 gene is restricted to neural crest cells migrating to the third branchial arch, apparently from r5, whereas this rhombomere contributes cells to both the second and third arches. We examined in the chick embryo how this spatially restricted expression is established. Expression occurs in precursors in both r5 and r6, and we show by cell labelling that both rhombomeres contribute to Krox‐20‐expressing neural crest, emigration occurring first from r6 and later caudally from r5. Krox‐20 transcripts are not detected in some precursors in rostral r5, presaging the lack of expression in cells migrating rostrally from this rhombomere. After transposition of r6 to the position of r4 or r5, many Krox‐20‐expressing cells migrate rostral to the otic vesicle, whereas when r5 is transplanted to the position of r4, only a small number of migrating cells express Krox‐20. These results indicate that, in the chick, Krox‐20 expression in branchial neural crest does not correlate with rhombomeric segmentation, and that there may be intrinsic differences in regulation between the r5 and r6 Krox‐20‐expressing populations.