Ultrastructure and composition of basement membrane separating mature ameloblasts from enamel

Abstract

At a late stage of amelogenesis, a basement-membrane-like (BML) structure appears between mature ameloblasts and the enamel surface. Although this BML structure is known to contain certain basement membrane components, its detailed nature and role were not well defined. As such, this study examined the BML structure using high-resolution electron microscopy combined with immunohistochemical staining. Mandibular rat incisors were processed for the preparation of Epon sections for ultrastructural observations, and frozen sections were used for immunostaining laminin, heparan sulphate proteoglycan (HSPG) and type IV collagen. The BML structure was characterized by the presence of abundant ribbon-like ‘double tracks’, 4.5–5.0 nm wide; the form known to be taken by HSPG in basal laminae. The main ultrastructural component of basal laminae, known as ‘cords’, was replaced by fine filaments of type IV collagen. Immunohistochemical staining of the BML structure showed an intense reaction for HSPG, moderate staining for type IV collagen and negligible staining for laminin. These observations indicate that this structure is an atypical basement membrane in which the cord network is replaced by type IV collagen filaments. However, the BML structure was found to be unusually rich in HSPG, similar to kidney glomerular basement membrane. It is likely that this specialized basement membrane mediates firm attachment of mature ameloblasts to the enamel surface, and filters the influx and efflux of materials to and from enamel during maturation.