Inhibition of sonic hedgehog signaling in vivo results in craniofacial neural crest cell death.
BACKGROUND
Sonic hedgehog (Shh) is well known for its role in patterning tissues, including structures of the head. Haploinsufficiency for SHH in humans results in holoprosencephaly, a syndrome characterized by facial and forebrain abnormalities. Shh null mice have cyclopia and loss of branchial arch structures. It is unclear, however, whether these phenotypes arise solely from the early function of Shh in patterning midline structures, or whether Shh plays other roles in head development.


RESULTS
To address the role of Shh after floorplate induction, we inhibited Shh signaling by injecting hybridoma cells that secrete a function-blocking anti-Shh antibody into the chick cranial mesenchyme. The antibody subsequently bound to Shh in the floorplate, notochord, and the pharyngeal endoderm. Perturbation of Shh signaling at this stage resulted in a significant reduction in head size after 1 day, loss of branchial arch structures after 2 days, and embryos with smaller heads after 7 days. Cell death was significantly increased in the neural tube and neural crest after 1 day, and neural crest cell death was not secondary to the loss of neural tube cells.


CONCLUSIONS
Reduction of Shh signaling after neural tube closure resulted in a transient decrease in neural tube cell proliferation and an extensive increase in cell death in the neural tube and neural crest, which in turn resulted in decreased head size. The phenotypes observed after reduction of Shh are similar to those observed after cranial neural crest ablation. Thus, our results demonstrate a role for Shh in coordinating the proliferation and survival of cells of the neural tube and cranial neural crest.
Ahlgren
S C
SC
Division of Biology, Beckman Institute MC 139-74, California Institute of Technology, Pasadena, 91125, USA. ahlgren@caltech.edu
Bronner-Fraser
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016923:Cell Death; D002455:Cell Division; D019070:Cell Lineage; D002465:Cell Movement; D002642:Chick Embryo; D005145:Face / Q000196:embryology*; D006257:Head / Q000196:embryology*; D053823:Hedgehog Proteins; D016142:Holoprosencephaly / Q000196:embryology*; D006825:Hybridomas / Q000378:metabolism / Q000637:transplantation; D008648:Mesoderm / Q000473:pathology; D009024:Morphogenesis / Q000235:genetics; D009432:Neural Crest / Q000254:growth & development*; D009436:Neural Tube Defects / Q000196:embryology*; D010641:Phenotype; D011506:Proteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D012249:Rhombencephalon / Q000196:embryology; D015534:Trans-Activators
D000911:Antibodies, Monoclonal; D053823:Hedgehog Proteins; D011506:Proteins; D015534:Trans-Activators