Enamelin gene expression during fetal and neonatal rabbit tooth organogenesis

Epithelial differentiation is a complex process which requires an integrated synthesis of DNA along with synthesis of a full complement of unique mRNAs and their respective proteins characteristic for each cell type. The time of initial transcription of enamel protein mRNAs and subsequent translation of proteins characteristic for secretory ameloblasts is not known. In order to determine when enamel protein mRNAs appear during New Zealand White rabbit molar tooth organogenesis, and when nascent enamel proteins are first translated, we analyzed early cap stages through late crown stages of molar tooth formation (i.e., 21-days gestation through 2-days postnatal). The biochemical phenotype which characterized rabbit ameloblasts were the acidic glycoproteins termed enamelins. Polyclonal antibodies were produced against the major fetal rabbit enamelin of approximately 70,000 daltons. Immunoprecipitation of enamelins from mRNA-directed translation products in a reticulocyte cell-free system, was used to characterize enamelin mRNAs. Enamelin mRNAs were first detected during bell stages (circa 23-days gestation), and persisted till crown stage (circa 28-days gestation). Indirect immunofluorescent localization of enamelin antigen showed staining over the extracellular enamel organ matrix by 23-days gestation. Neither enamelin mRNAs or polypeptides were detected during early or late cap stages of odontogenesis. Transcription of enamelin mRNAs coding for two enamelins of 65 an 58 kd (kilodaltons) appeared to be closely coupled with the translation of these enamel proteins. We assume that close-range ectomesenchyme- derived instructions mediate the biochemical differentiation of ameloblasts between 21-days and 23-days gestation during fetal rabbit development.