Serotonergic 5-HT(2B) receptor controls tissue-nonspecific alkaline phosphatase activity in osteoblasts via eicosanoids and phosphatidylinositol-specific phospholipase C.
In previous studies, we observed that mice knocked out for the serotonin-2B receptor (5-HT(2B)R) show defects in bone homeostasis. The present work focuses on the downstream targets relaying the anabolic function of this receptor in osteoblasts. A functional link between the 5-HT(2B)R and the activity of the tissue-nonspecific alkaline phosphatase (TNAP) is established using the C1 osteoprogenitor cell line. During C1 osteogenic differentiation, both 5-HT(2B)R and TNAP mRNA translations are delayed with respect to extracellular matrix deposition. Once the receptor is expressed, it constitutively controls TNAP activity at a post-translational level along the overall period of mineral deposition. Indeed, pharmacological inhibition of the 5-HT(2B)R intrinsic activity or shRNA-mediated 5-HT(2B)R knockdown prevents TNAP activation, but not its mRNA translation. In contrast, agonist stimulation of the receptor further increases TNAP activity during the initial mineralization phase. Building upon our previous observations that the 5-HT(2B)R couples with the phospholipase A2 pathway and prostaglandin production at the beginning of mineral deposition, we show that the 5-HT(2B)R controls leukotriene synthesis via phospholipase A2 at the terminal stages of C1 differentiation. These two 5-HT(2B)R-dependent eicosanoid productions delineate distinct time windows of TNAP regulation during the osteogenic program. Finally, prostaglandins or leukotrienes are shown to relay the post-translational activation of TNAP via stimulation of the phosphatidylinositol-specific phospholipase C. In agreement with the above findings, primary calvarial osteoblasts from 5-HT(2B)R-null mice exhibit defects in TNAP activity.
Baudry
Anne
A
Cellules Souches, Signalisation et Prions, INSERM U747, Université Paris Descartes, Paris, France.
Bitard
Juliette
J
Mouillet-Richard
Sophie
S
Locker
Morgane
M
Poliard
Anne
A
Launay
Jean-Marie
JM
Kellermann
Odile
O
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D002113:Calcification, Physiologic; D002454:Cell Differentiation; D002478:Cells, Cultured; D015777:Eicosanoids / Q000378:metabolism*; D015289:Leukotrienes / Q000096:biosynthesis; D051379:Mice; D010006:Osteoblasts / Q000201:enzymology / Q000378:metabolism*; D010012:Osteogenesis; D054801:Phosphoinositide Phospholipase C / Q000378:metabolism*; D044403:Receptor, Serotonin, 5-HT2B / Q000502:physiology*; D013997:Time Factors
D015777:Eicosanoids; D015289:Leukotrienes; D044403:Receptor, Serotonin, 5-HT2B; D000469:Alkaline Phosphatase; D054801:Phosphoinositide Phospholipase C