Takashi Fujita, Ryo Fukuyama, Nobuo Izumo, Takao Hirai, Toru Meguro, Hiromichi Nakamuta and Masao Koida*
Department of Pharmacology, Faculty of Pharmaceutical Sciences, Setsunan University, Nagaotohgecho 45-1, Hirakata, Osaka 573-0101, Japan
*Corresponding author. FAX: +81-72-866-3108, E-mail: koida@pharm.setsunan.ac.jp
Abstract: During 28-day culture of bone marrow- and calvaria-derived osteoblasts, the constant presence of parathyroid hormone (PTH)(1 - 34) retarded differentiation and nodule formation (NF) in a dose-dependent fashion (C-phase). In contrast, addition of PTH(1 - 34) in late stage cultures (from day 10 to 21) accelerated NF (A-phase). The stable production of such an A-phase allowed us to study the mechanism of bone anabolic action of PTH(1 - 34). Subcellular localization studies of core binding factor a1 (Cbfa1) and reporter assays provided the results indicating that in the A-phase, PTH(1 - 34) triggers its bone anabolic action via enhancement of Cbfa1 transactivation. RT-PCR and Northern blot analyses revealed that alkaline phosphatase, osteocalcin and bone sialoprotein expression decreased in the C-phase and increased in the A-phase; however, expression of other bone proteins (Cbfa1, PTH/PTH-related peptide-receptor, osteopontin, collagen I a1, collagen I a2, vitamin K-dependent g-glutamyl carboxylase) did not change in a phase transition-related manner. Ovariectomized osteopenic mice, treated with PTH(1 - 34) (4 and 40 mg/kg, s.c., every other day, 4 or 6 weeks), recovered lost bone, displayed elevated nuclear localization of Cbfa1 in tibiae without alteration of its cytosolic level and exhibited upregulation of expressions of the same set of proteins (alkaline phosphatase, osteocalcin and bone sialoprotein) in femora. These results obtained by a concerted study in vitro and in vivo suggest that PTH triggers its osteogenic action via promotion of the transactivation of Cbfa1.
Keywords: Parathyroid hormone, Osteogenesis, Osteoblast, Core binding factor a1, Transactivation
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