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Hyper-expression of Osteocalcin mRNA in Odontoblasts of Hyp Mice

¹ Department of Pediatric Dentistry, Osaka University Graduate School of Dentistry, 1-8, Yamadaoka, Suita, Osaka 565-0871, Japan; and
² Division of Pediatric Dentistry, Department of Developmental and Reconstructive Medicine, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1, Sakamoto, Nagasaki, 852-8588, Japan;

View larger version (146K): [in a new window]   Figure 1. Photomicrographs of lower first molars on PN6d. (a) WT mouse. (b) Hyp mouse. Widened predentin can be seen. (c) OC mRNA expression in WT mouse specimen. Reactivity is observed in odontoblasts (arrowheads), which weakened with the progress of the cytodifferentiation and mineralization processes (large arrowheads). (d) OC mRNA expression in Hyp mouse. The distribution of signals (arrowheads) is similar to that of WT mice. (a,b) HE staining; (c,d) in situ hybridization. AB, ameloblasts; D, dentin; E, enamel; OB, odontoblasts; PD, predentin; PO, pre-odontoblasts.

View larger version (24K): [in a new window]   Figure 2. Comparison of the quantities of OC, OSN, OPN, DMP1, Col I, and ß-actin mRNA in tooth germs of Hyp and WT mice. (A) Electrophoresis of PCR products for OC, OSN, OPN, DMP1, ColI, and ß-actin with PN6d WT mice tooth germs. Single bands of the expected size for each amplification product are shown. (B) Relative quantities of mRNA (mean and SEM) for OC, OSN, OPN, DMP1, and ColI in tooth germs on PN2d and PN6d. Significant differences in the amount of OC mRNA expression between Hyp and WT mice were seen on both post-natal days (*P < 0.05; Mann-Whitney U test).

View larger version (173K): [in a new window]   Figure 3. Photomicrographs of tooth germs removed from mice on E17d (a-f) and tooth germs cultured for 10 days after E17d (E17d-C10) (g-l). (a-c, g-i) WT mouse. (d-f, j-l) Hyp mouse. (b,e,h,k) Higher magnifications of the boxed areas in (a,d,g,j), respectively. (c,f,i,l) In situ hybridization. (a-f) Polarized odontoblasts cannot be seen. (c,f) No reactivity for OC mRNA is shown. (g-l) Most of the odontoblasts have differentiated into the secretory phase, and thin dentin matrix can be observed [between arrowheads in (h) and (k)]. (i,l) Reactivity for OC mRNA in odontoblasts. AB, ameloblasts; D, dentin; DP, dental papilla; EO, enamel organ; OB, odontoblasts; PA, pre-ameloblasts; PO, pre-odontoblasts.

View larger version (11K): [in a new window]   Figure 4. Relative quantities of mRNA (mean and SEM) for OC, OSN, OPN, DMP1, and ColI in tooth germs cultured for 10 days after E17d. Significant differences in the amount of OC mRNA expression between Hyp and WT mice are shown (**P < 0.01; Mann-Whitney U test).