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We studied the effects of PTH on the expression of tumor necrosis factor-related activation-induced cytokine (TRANCE), osteoprotegerin (OPG), and receptor activator of NF kappaB (RANK) messenger RNA (mRNA) in cultured murine bone marrow, calvaria, and osteoblasts. TRANCE, OPG, and RANK are recently identified regulators of osteoclast formation. Bone marrow cells were cultured with or without PTH(1-34) for 6 days. TRANCE, OPG, and RANK mRNA were measured by RT-PCR. In 6-day cultures, PTH stimulated the number of OCL/well in a dose-dependent manner. A time course showed significant (P &lt; 0.01) increases in OCL/well after 24 h of PTH (100 ng/ml). TRANCE mRNA expression, like OCL formation, increased dose dependently and was maximal, with 10-100 ng/ml PTH. In contrast, OPG mRNA expression was decreased by 0.1 ng/ml PTH (40%) and completely abolished by 1 ng/ml. TRANCE mRNA expression was rapidly stimulated by PTH (maximal response at 1 h, 8.1-fold over control). Expression declined by 40% at 24 h but was still much greater than control at 6 days (4.6-fold) in a time-course study. PTH caused a transient stimulation of OPG mRNA at 1 h (2-fold), which returned to basal levels by 2 h. After 6 h, PTH completely inhibited OPG mRNA. There were only minor effects of PTH on RANK mRNA expression. PTH had less potent effects on TRANCE and OPG mRNA expression in calvaria organ cultures and osteoblasts. In mouse calvaria cultures, TRANCE expression was detectable in controls and was increased 2.9-fold by PTH at 24 h. PTH treatment of calvaria decreased OPG expression by 30% at 6 h. MC3T3 E-1 osteoblastic cells expressed minimal levels of TRANCE mRNA either before or after PTH treatment. OPG mRNA was present in MC3T3 E-1 cells, but levels were not modulated by PTH. In primary osteoblastic cells, PTH stimulated TRANCE mRNA expression 4-fold at 2 h and inhibited OPG mRNA expression by 46%. These results demonstrate a tight correlation between the ability of PTH to stimulate OCL formation in marrow culture and expression of TRANCE (r = 0.87, P &lt; or = 0.05) and OPG mRNA (r = -0.88, P &lt; or = 0.05). Reciprocal regulation of TRANCE and OPG mRNA by PTH preceded its effects on OCL formation by 18-23 h. Hence, it is likely that PTH regulates bone resorption, at least in part, via its effects on TRANCE and OPG expression.

endocrinology

Parathyroid Hormone Stimulates TRANCE and Inhibits Osteoprotegerin Messenger Ribonucleic Acid Expression in Murine Bone Marrow Cultures: Correlation with Osteoclast-Like Cell Formation*