Absence of neural crest cells from the region surrounding implanted notochords in situ

Avian neural crest cells migrating along the trunk ventral pathway are distributed throughout the rostral half of the sclerotome with the exception of a neural crest cell-free space of approximately 85 microns width surrounding the notochord. To determine if this neural crest cell-free space results from the notochord inhibiting neural crest cell migration, a length of quail notochord was implanted lateral to the neural tube along the neural crest ventral migratory pathway of 2-day chicken embryos. The subsequent distribution of neural crest cells was analyzed in embryos fixed 2 days after grafting. When the donor notochord was isolated using collagenase, neural crest cells avoided the ectopic notochord and were absent from the area immediately surrounding the implant (mean distance of 43 microns). The neural crest cell-free space was significantly less when notochords were isolated using trypsin or chondroitinase digestion and was completely eliminated when notochords were fixed with paraformaldehyde or methanol prior to implantation. The implanted notochords did not appear to affect the overall number of neural crest cells, and therefore were unlikely to exert this effect by altering their viability. These results suggest that the notochord produces a substance that can inhibit neural crest cell migration and that this substance is trypsin and chondroitinase labile.