Enamel Defects Associated With Dentin Sialophosphoprotein Mutation in Mice

Dentin sialophosphoprotein (DSPP) is an extracellular matrix protein that is highly expressed in odontoblasts, but only transiently expressed in presecretory ameloblasts during tooth development. We previously generated a knockin mouse model expressing a mouse equivalent (DSPP, p.P19L) of human mutant DSPP (p.P17L; referred to as “Dspp(P19L/+)”), and reported that Dspp(P19L/+) and Dspp(P19L/P19L) mice manifested a dentin phenotype resembling human dentinogenesis imperfecta (DGI). In this study, we analyzed pathogenic effects of mutant P19L-DSPP on enamel development in Dspp(P19L/+) and Dspp(P19L/P19L) mice. Micro-Computed Tomography (μCT) analyses of 7-week-old mouse mandibular incisors showed that Dspp(P19L/P19L) mice had significantly decreased enamel volume and/or enamel density at different stages of amelogenesis examined. Acid-etched scanning electron microscopy (SEM) analyses of mouse incisors demonstrated that, at the mid-late maturation stage of amelogenesis, the enamel of wild-type mice already had apparent decussating pattern of enamel rods, whereas only minute particulates were found in Dspp(P19L/+) mice, and no discernible structures in Dspp(P19L/P19L) mouse enamel. However, by the time that incisor enamel was about to erupt into oral cavity, distinct decussating enamel rods were evident in Dspp(P19L/+) mice, but only poorly-defined enamel rods were revealed in Dspp(P19L/P19L) mice. Moreover, μCT analyses of the mandibular first molars showed that Dspp(P19L/+) and Dspp(P19L/P19L) mice had a significant reduction in enamel volume and enamel density at the ages of 2, 3, and 24weeks after birth. Backscattered and acid-etched SEM analyses revealed that while 3-week-old Dspp(P19L/+) mice had similar pattern of enamel rods in the mandibular first molars as age-matched wild-type mice, no distinct enamel rods were observed in Dspp(P19L/P19L) mice. Yet neither Dspp(P19L/+) nor Dspp(P19L/P19L) mice showed well-defined enamel rods in the mandibular first molars by the age of 24weeks, as judged by backscattered and acid-etched SEM. In situ hybridization showed that DSPP mRNA level was markedly reduced in the presecretory ameloblasts, but immunohistochemistry revealed that DSP/DSPP immunostaining signals were much stronger within the presecretory ameloblasts in Dspp mutant mice than in wild-type mice. These results suggest that mutant P19L-DSPP protein caused developmental enamel defects in mice, which may be associated with intracellular retention of mutant DSPP in the presecretory ameloblasts.