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• W2024884314 abstract "PTH has anabolic and catabolic effects in bone through activation of the PTH-1 (PTH/PTHrP) receptor and the cAMP/protein kinase A pathway. The effects of agents that regulate cAMP in nontransformed osteoblasts in relation to cell differentiation have not been described. The purpose of this study was to determine the effects of PTH fragments with differing cAMP-stimulating activity, and nonPTH cAMP regulators on PTH-1 receptor expression and activity, and osteoblast differentiation in vitro using MC3T3-E1 and primary rat calvarial cells. PTH (1–34), but not PTH (53–84), (7–34), or PTHrP (107–139) treatment (24 h) resulted in down-regulation of steady-state messenger RNA for the PTH-1 receptor. Forskolin (a stimulator of cAMP accumulation) also down regulated the PTH-1 receptor, whereas 9-(tetrahydro-2-furyl) adenine (THFA) (an inhibitor of adenylyl cyclase) had no effect. Similarly, PTH (1–34) treatment for 48 h abolished PTHrP binding to cell surface receptors; however, neither the PTH analogs nor the cAMP regulating agents altered PTH binding or numbers of binding sites on osteoblastic cells. Basal levels of cAMP were reduced in cultured cells treated for 6 days with PTH (7–34) or THFA compared with controls. In contrast, PTH-stimulated cAMP levels were significantly increased in cultures treated with PTH (7–34) and THFA for 6 days during osteoblast differentiation and were decreased in cultures treated with PTH (1–34) and forskolin compared with controls. To evaluate effects of the cAMP pathway on osteoblast differentiation, cultures were treated continuously with PTH analogs and cAMP regulators during an 18-day differentiation regime, total RNA was isolated at multiple time points, and Northern blot analysis for osteocalcin (OCN) was performed. THFA and PTH (7–34)-treated cultures had increased OCN expression; whereas, PTH (1–34) and forskolin reduced OCN expression. Interestingly, PTH (7–34) and THFA-treated cultures had increased mineralized nodule formation, in contrast to PTH (1–34) and forskolin treatment, which reduced nodule formation. Similarly, calcium accumulation in cultures was significantly increased in the PTH (7–34) and THFA-treated cultures and reduced in the PTH (1–34) and forskolin-treated cultures. These data demonstrate that agents that increase cAMP down regulate PTH-1 receptor messenger RNA and inhibit osteoblast differentiation in vitro. Agents that reduce or block adenylyl cyclase or cAMP activity do not alter PTH-1 receptor expression or binding, but have striking effects on promoting osteoblast differentiation. We conclude that many effects of PTH on osteoblasts may be mimicked or antagonized by agents that alter cAMP activity and bypass the PTH-1 receptor." @default.
• W2024884314 created "2016-06-24" @default.
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• W2024884314 date "1999-07-01" @default.
• W2024884314 modified "2023-10-14" @default.
• W2024884314 title "3′,5′-Cyclic Adenosine Monophosphate Activation in Osteoblastic Cells: Effects on Parathyroid Hormone-1 Receptors and Osteoblastic Differentiation in Vitro*" @default.
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• W2024884314 doi "https://doi.org/10.1210/endo.140.7.6872" @default.
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