Expression, localisation and synthesis of versican by the enamel organ of developing mouse molar tooth germ: An in vivo and in vitro study

Objective: Versican is a large, aggregating chondroitin sulphate proteoglycan. In dental tissue, versican expression occurs primarily in mesenchymal tissue but rarely in epithelial tissue. We investigated the expression, localisation and synthesis of versican in the enamel organ of the developing tooth germ. Design: To elucidate versican localisation in vivo, in situ hybridisation and immunohistochemistry were conducted in foetal ICR mice at E11.5-E18.5. Epithelium and mesenchyme from the lower first molars at E16.0 were enzymatically separated and versican mRNA expression was investigated by semi-quantitative RT-PCR. Organ culture of the separated samples combined with metabolic labelling with [S-35], followed by gel filtration, was performed to analyse secreted proteoglycans. Results: Versican mRNA was first expressed in the thickened dental epithelium at E12.0 and continued to be expressed in the enamel organ until the bell stage. Versican immunostaining was detected in the stellate reticulum areas from the bud stage to the apposition stage. The enamel organ at E16.0 expressed versican mRNA at a level comparable to that in dental mesenchyme. Furthermore, when compared to dental mesenchyme, about 1/2-3/4 of the [S-35]-labelled versican-like large proteoglycan was synthesised and released into tissue explants by the enamel organ. Conclusions: The dental epithelium of developing tooth germ is able to synthesise significant amounts of versican. (C) 2010 Elsevier Ltd. All rights reserved.