Reversible suppression of in vitro biomineralization by activation of protein kinase A.
Parathyroid hormone (PTH-(1-34)) potently suppresses apatite deposition in osteoblastic cultures. These inhibitory effects are mediated through signaling events following PTH receptor binding. Using both selective inhibitors and activators of protein kinase A (PKA), this study shows that a transient activation of PKA is sufficient to account for PTH's inhibition of apatite deposition. This inhibition is not a result of reduced cell proliferation, reduced alkaline phosphatase activity, increased collagenase production, or lowering medium pH. Rather, data suggest a functional relationship between matrix assembly and apatite deposition in vitro. Bone sialoprotein (BSP) and apatite co-localize in the extracellular matrix of mineralizing cultures, with matrix deposition of BSP temporally preceding that of apatite. Transient activation of PKA by either PTH-(1-34) or short term cAMP analog treatment blocks the deposition of BSP in the extracellular matrix without a significant reduction in the total amount of BSP synthesized and secreted. This effect is reversible after allowing the cultures to recover in the absence of PKA activators for several days. Thus, a transient activation of PKA may suppress mineral deposition in vitro as a consequence of altering the assembly of an extracellular matrix permissive for apatite formation.
Wang
A
A
Department of Biomedical Engineering, The Lerner Research Institute of The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Martin
J A
JA
Lembke
L A
LA
Midura
R J
RJ
D000469:Alkaline Phosphatase / Q000378:metabolism; D001031:Apatites / Q000378:metabolism; D002113:Calcification, Physiologic / Q000187:drug effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D058574:Integrin-Binding Sialoprotein; D010281:Parathyroid Hormone / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D012795:Sialoglycoproteins / Q000032:analysis / Q000235:genetics
D001031:Apatites; D058574:Integrin-Binding Sialoprotein; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D000469:Alkaline Phosphatase