Gene expression and localization of amelogenin were studied in the developing rat incisor by the methods of in situ hybridization and immunohistochemistry. ISH revealed the first expression of amelogenin mRNA in the inner enamel epithelium of the cervical loop. The signals were clearly observed in pre-ameloblasts in the region bordering on predentin formation and became more intense toward the cells on the initial enamel matrix secretion. The maximal signals were found in the cytoplasm of secretory ameloblasts. From the terminal secretion zone, the signals then became gradually weaker toward the incisal edge but were still evident in the cytoplasm of shortening, transitional ameloblasts and those at the early maturation stage. No signals were found in the cells of the stratum intermedium and stellate reticulum throughout amelogenesis. Immunohistochemistry by means of an antibody against amelogenin C-telopeptide consisting of 12 amino acids revealed immunoreaction in the secretory ameloblasts reacting to the ISH. When a polyclonal antibody against amelogenin was used, immunoreaction was found in the distal ends of ruffle-ended ameloblasts (RA) in the maturation zone. Those results indicated that amelogenin is synthesized by ameloblastic cells from the inner enamel epithelium to the early maturation stage and is then resorbed by the RA.

Morphological differentiation of the distal ends of ameloblasts (AMs) from the late presecretory to early secretory zones of the rat upper incisor was studied by electron microscopy. Preameloblasts (PAs) showed a high columnar shape, with the nucleus located in the basal portion. The Golgi apparatus occupied the supranuclear region with type-1 vesicles, and microvilli were present at the distal cell membrane. Coarse-textured material was observed inside the type-1 vesicles and in the lateral intercellular spaces as well as along the distal cell membrane, whereas fine-textured material was found along the distal cell membrane. Near the region of initial enamel formation, large matrix islands were found in the lateral intercellular space. A thin electron-dense layer was observed on the dentin surface. This layer might have occurred as a result of diffusion after degradation of the coarse-or fine-textured material into a finer substance in the extracellular spaces. In the region of initial enamel formation, the distal cell membrane of AMs was flat, but shallow and narrow membrane invaginations were associated with the cell membrane close to the matrix islands. In the region of inner enamel formation, a cone-shaped Tomes' process was formed between large matrix islands which had developed in the intercellular spaces between the lateral portions of the distal ends of AMs. It was considered that the membrane invaginations which had existed at the distal end of PAs moved lower toward the distal terminal bar, thereafter becoming microvilli.

The multi-functional protein osteonectin is widely distributed throughout many tissues. It plays a role in tissue morphogenesis and affects the interaction between cells and the extracellular matrix.

No abstract