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Fluid flow induces Rankl expression in primary murine calvarial osteoblasts
Author(s) -
Mehrotra Meenal,
Saegusa Masatomo,
Wadhwa Sunil,
Voznesensky Olga,
Peterson Donald,
Pilbeam Carol
Publication year - 2006
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.20864
Subject(s) - rankl , protein kinase c , osteoclast , mapk/erk pathway , activator (genetics) , chemistry , osteoprotegerin , phosphorylation , receptor , protein kinase a , messenger rna , microbiology and biotechnology , medicine , endocrinology , biology , biochemistry , gene
Mechanical loading of bone generates fluid flow within the mineralized matrix that exerts fluid shear stress (FSS) on cells. We examined effects of FSS on receptor activator of nuclear factor κ B ligand (RANKL), a critical factor for osteoclast formation. Primary murine osteoblasts were subjected to pulsatile FSS (5 Hz, 10 dynes/cm 2 ) for 1 h and then returned to static culture for varying times (post‐FSS). Protein levels were measured by Western analysis and mRNA by Northern analysis, RT‐PCR and quantitative PCR. There were 20‐ to 40‐fold increases in RANKL mRNA at 2–4 h post‐FSS. RANKL protein was induced by 2 h post‐FSS and remained elevated for at least 8 h. Effects were independent of cyclooxygenase‐2 activity. Small increases (up to three‐fold) in mRNA of the decoy receptor for RANKL, osteoprotegerin, were seen. Five min of FSS, followed by static culture, was as effective in stimulating RANKL mRNA as 4 h of continuous FSS. FSS induced cAMP activity, and H‐89, a protein kinase A (PKA) inhibitor, blocked the FSS induction of RANKL. H‐89 also inhibited the PKC pathway, but specific PKC inhibitors, GF109203X and Go6983, did not inhibit FSS‐induced RANKL. FSS induced phosphorylation of ERK1/2, and PD98059, an inhibitor of the ERK pathway, inhibited the FSS induction of RANKL mRNA 60%–90%. Thus, brief exposure to FSS resulted in sustained induction of RANKL expression after stopping FSS, and this induction was dependent on PKA and ERK signaling pathways. Increased RANKL after mechanical loading may play a role in initiating bone remodeling. J. Cell. Biochem. 98: 1271–1283, 2006. © 2006 Wiley‐Liss, Inc.