An in vitro model for the study of the role of innervation in circumvallate papillae morphogenesis.

The following study was done to demonstrate the reliability of an in vitro model for use in the study of early events and the role of innervation in mouse circumvallate papillae development. Gestational day (gd)-11 fetuses were partially dissected to produce explants that included the mandibular, hyoid, third and fourth branchial arches and their ganglia. In ganglionectomized explants, the nodose ganglia and either the geniculate, petrosal or both ganglia were removed. Explants were cultivated in roller tube culture for 24, 48, 72, and 96 h of culture and examined for the presence of papillary structures. Innervation was verified by immunostaining for neural cell adhesion molecule (NCAM). In all control explants, circumvallate papillae had formed by 72 h in culture. These papillae were innervated by fibers originating in petrosal or nodose ganglia, although, in a small number, fibers from the geniculate also contributed. Circumvallate papillae also formed in some explants in which either the geniculate or petrosal ganglia had been removed. However, placodal structures failed to mature into papillary structures even by 96 h in explants in which both ganglia had been removed. Our results demonstrate that an in vitro model using branchial arch explants supports the morphogenesis of an epithelial placode through the formation of a definite papillary structure, the circumvallate papilla, with an integrated nerve. Our results also indicate that, whereas the initial stages in gustatory papillae formation, the formation of a placode, are nerve-independent, the maturation of the placodal structure to form a papilla requires the presence of an intact nerve.