Document Type
Article
Language
eng
Format of Original
9 p.
Publication Date
4-2008
Publisher
Elsevier
Source Publication
Bone
Source ISSN
8756-3282
Abstract
To determine the role of Ca2+ signaling in activation of the Mitogen-Activated Protein Kinase (MAPK) pathway, we subjected MC3T3-E1 pre-osteoblastic cells to inhibitors of Ca2+ signaling during application of fluid shear stress (FSS). FSS only activated ERK1/2, rapidly inducing phosphorylation within 5 min of the onset of shear. Phosphorylation of ERK1/2 (pERK1/2) was significantly reduced when Cai2+ was chelated with BAPTA or when Ca2+ was removed from the flow media. Inhibition of both the L-type voltage-sensitive Ca2+ channel and the mechanosensitive cation-selective channel blocked FSS-induced pERK1/2. Inhibition of phospholipase C with U73122 significantly reduced pERK1/2. This inhibition did not result from blockage of intracellular Ca2+ release, but a loss of PKC activation. Recent data suggests a role of ATP release and purinergic receptor activation in mechanotransduction. Apyrase-mediated hydrolysis of extracellular ATP completely blocked FSS-induced phosphorylation of ERK1/2, while the addition of exogenous ATP to static cells mimicked the effects of FSS on pERK1/2. Two P2 receptors, P2Y2 and P2X7, have been associated with the anabolic responses of bone to mechanical loading. Using both iRNA techniques and primary osteoblasts isolated from P2X7 knockout mice, we found that the P2X7, but not the P2Y2, purinergic receptor was involved in ERK1/2 activation under FSS. These data suggest that FSS-induced ERK1/2 phosphorylation requires Ca2+-dependent ATP release, however both increased Cai2+ and PKC activation are needed for complete activation. Further, this ATP-dependent ERK1/2 phosphorylation is mediated through P2X7, but not P2Y2, purinergic receptors.
Recommended Citation
Liu, Dawei; Genetos, Damian C.; Shao, Ying; Geist, Derik J.; Li, Jiliang; Ke, Hua Zhe; Turner, Charles H.; and Duncan, Randall L., "Activation of Extracellular-signal Regulated Kinase (ERK1/2) by Fluid Shear is Ca2+- and ATP-dependent in MC3T3-E1 Osteoblasts" (2008). School of Dentistry Faculty Research and Publications. 71.
https://epublications.marquette.edu/dentistry_fac/71
Comments
Accepted version. Bone, Vol. 42, No. 4 (April 2008): 644-652. DOI. © 2008 Elsevier. Used with permission.
NOTICE: this is the author’s version of a work that was accepted for publication in Bone. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Bone, VOL 42, ISSUE 4, April 2008, DOI.