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Phex Mutation Causes the Reduction of Npt2b mRNA in Teeth

Department of Pediatric Dentistry, Osaka University Graduate School of Dentistry, 1-8, Yamadaoka, Suita, Osaka 565-0871, Japan

View larger version (120K): [in this window] [in a new window]   Figure 1. Expression of Npt2 mRNA in wild-type mice, with the use of RT-PCR (a), Northern blot (b), and in situ hybridization (c–g) analyses. (a) Single bands of the expected size for the amplification products of Npt2b were observed in tooth germs from mice aged 2, 6, and 10 days. (b) A strong expression of Npt2b mRNA can be seen in the tooth germ specimens. (c) Photomicrograph of a distal portion of a lower first molar. (d) Higher magnification of the boxed area in (c). Mature ameloblasts (MA) show the signals. (e) Adjacent section of (d). H-E staining. (f) Higher magnification of the boxed area in (c). Signals can be seen in secretory ameloblasts (SA) and odontoblasts (arrows). (g) Section adjacent to (e). H-E staining. D, dentin; E, enamel; OB, odontoblasts.

View larger version (18K): [in this window] [in a new window]   Figure 2. Comparisons of quantities of Npt2b mRNA in Hyp and wild-type mice (n = 10 of each). (a) Relative quantities of mRNA (mean ± SEM) for Npt2b in teeth and intestines of mice at the age of 6 days. Significant differences between Hyp and wild-type mice were found. (b) Relative quantities of mRNA (mean ± SEM) for Npt2b in cultured tooth germs. The expression level in cultured tooth germs derived from Hyp mice was significantly lower than that in those from wild-type mice (*P < 0.05, **P < 0.01, Mann-Whitney U-test).

View larger version (161K): [in this window] [in a new window]   Figure 3. Photomicrographs of tooth germs derived from mice on embryonic day 17 and cultured for 14 days. (a) Tooth germs cultured without oligo-deoxynucleotide. (b) Higher magnification of the boxed area in (a). (c) Tooth germs cultured with sense oligo-deoxynucleotide for the Phex. (d) Higher magnification of the boxed area in (c). (e) Tooth germs cultured with antisense oligo-deoxynucleotide for the Phex gene. (f) Higher magnification of the boxed area in (e). AB, ameloblasts; D, dentin; DP, dental papilla; EO, enamel organ; OB, odontoblasts.

View larger version (14K): [in this window] [in a new window]   Figure 4. Comparisons of quantities of Phex and Npt2b mRNA in cultured tooth germs. (a) Relative quantities of Phex mRNA (mean ± SEM) in tooth germs cultured with or without oligo-deoxynucleotide (n= 8 of each). The expression level in tooth germs cultured with antisense oligo-deoxynucleotide was significantly lower than that of tooth germs cultured with sense oligo-deoxynucleotide and without any oligo-deoxynucleotides (*P < 0.01, Mann-Whitney U-test). (b) Relative quantities of Npt2b mRNA (mean ± SEM) in tooth germs cultured with or without oligo-deoxynucleotide (n = 8 of each). The expression level in tooth germs cultured with antisense ODN was significantly lower than that in tooth germs cultured with sense oligo-deoxynucleotide and without oligo-deoxynucleotide (*P < 0.01, Mann-Whitney U-test).