Spatial and temporal activity of the dentin sialophosphoprotein gene promoter: differential regulation in odontoblasts and ameloblasts

Dentin sialoprotein and dentin phosphoprotein are non-collagenous proteins that are cleavage products of dentin sialophosphoprotein (DSPP). Although these two protein products are believed to have a crucial role in the process of tooth mineralization, their precise biological functions and the molecular mechanisms of gene regulation are not clearly understood. To understand such functions, we have developed a transgenic mouse model expressing a reporter gene (lacZ) under the control of approximately 6 kb upstream sequences of Dspp. The transgenic fusion protein was designed to reside within the cells to facilitate the precise identification of cell type and developmental stages at which the Dspp-lacZ gene is expressed. The results presented in this report demonstrate: (a) the 6 kb upstream sequences of Dspp have the necessary regulatory elements to direct the tissue specific expression of the transgene similar to endogenous Dspp, (b) both odontoblasts and ameloblasts exhibit transgene expression in a differentiation dependent manner, and (c) a differential regulation of the transgene in odontoblasts and ameloblasts occurs during tooth development and mineralization.     

Tissue-specific expression of dentin sialophosphoprotein (DSPP) and its polymorphisms in mouse tissues

Dentin sialophosphoprotein (DSPP) consists of dentin sialoprotein (DSP) and dentin phosphoprotein (DPP). DSPP is highly expressed in mineralized tissues. However, recent studies have shown that DSPP is also expressed in several active metabolic ductal epithelial tissues and exists in a variety of sequences. We have investigated DSPP expression in various mouse tissues using RT-PCR, in situ hybridization and immunohistochemical analyses. To identify DSPP gene polymorphisms, we screened a mouse tooth cDNA library as well as isolated and characterized DSPP variations. Our results show that DSPP is predominantly expressed in teeth and moderately in bone tissues. We also have characterized a full-length DSPP cDNA clone with an open-reading frame of 940 codons and this polyadenylation signal. Compared to previously reported mouse DSPP cDNAs, 13 sequence variations were identified, including 8 non-synonymous single nucleotide polymorphisms and an in-frame indel (8 amino acids) at DPP domain of the mouse DSPP. These 8 amino acids are rich in aspartic acid and serine residues. Northern blot assay showed a prominent band at 4.4 kb. RT-PCR demonstrated that this mouse DSPP gene was dominantly expressed in teeth. The predicted secondary structure of DPP domain of this DSPP showed differences from the previously published mouse DPPs, implying that they play different roles during tooth development and formation.