The study of neural crest cells by the mammalian whole embryo culture

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Mammlian neural crest cells have been investigated using morphological techniques with in vivo embryos or primary cell culture. Whole embryo culture (WEC) has made it possible to study the cells in more experimental way in which embryos are dossected out from their mother for manipulation. WEC can provide with appropriate method for developmental biology. In praticular, it has more advantages to use mouse embryos for wec since there are some mutants which have facial abmormality. In this current year, the following experiments were made to perturb the facial formation by chemical or operative treatment using mouse WEC from day 10 of gestation.1) Perturbation of facial formation with chytochalasin D (CD)C57BL/6 mouse embryos were exposed to 150ng/ml of CD for 2 hours using WEC. Facial mal-formation (cleft lip) was observed in 70.4% of the CD-treated embryos at the end point of the culture for 24 hours. The distribution of F-actin in nasal epithelium was observed to be partially broken in rhodamine-phalloidin staining image just after the treatment. Distrubed F-actin might be related to the cleft lip formation.2) Induction of cleft lip by early open yolk sacOrening of yolk sac (OYS) is indispensable to WEC. In C57BL/6 mouse embryos, when it has been made before 8-tail somite stage, cleft lip was induced to 100% of the embryos. Scanning electron microscopy of the presumptive fusion area in early OYS embryos showed that the disappearance of microvilli which were observed in mormal development was not occured, but that the surface of epithelium was spherical, and many debrises were observed.