Initial innervation of the palatal gustatory epithelium in the rat as revealed by growth-associated protein-43 (GAP-43) immunohistochemistry.

We studied the earliest stages of the palate in rat embryos using scanning electron microscopy and immunohistochemistry of growth-associated protein-43 (GAP-43) to investigate the role of nerves in the development of the palatal taste buds. Chronological sequences of the palatal gustatory structures revealed characteristic several stages: 1) At embryonic day 13.5 (E13.5), the palatal shelves were widely separated, and no nerves could be observed in the vicinity of their epithelium which was formed of an undifferentiated single cell layer. 2) At E14, intraepithelial GAP-43-immunoreactive fine nerves were first observed along the medial border of the palatal shelves which became several layers thick but still separate along their entire length. 3) At E15, the fusion process resulted in the formation of cranial parts of the soft palate, the epithelium of which was heavily innervated and revealed small fungiform-like papillae devoid of nerves. 4) As the fusion process continued more caudally at E15, there was a substantial increase in palatal innervation and number of fungiform-like papillae. Primordial stages of taste buds were first distinguished in the papillae where they coincided with sparsely distributed GAP-43-immunoreactive nerve fibers. 5) At E16, the whole soft palate was eventually differentiated and attained its definitive morphology. Different stages of taste buds (i.e. pored and non-pored) were recognized, and an extensive subgemmal plexus characteristic for the adult palatal taste buds was observed. 6) Mature taste buds with alpha-gustducin-immunopositive cells were observed at E18, and their numbers increased gradually with age. The present study reveals that the gustatory nerves preceded the development of taste buds in the palate of rats, and therefore may have some roles in the initial induction of taste buds as proposed in lingual taste buds.