The role of actin cytoskeleton in oscillatory fluid flow-induced signaling in MC3T3-E1 osteoblasts.
Fluid flow due to loading in bone is a potent mechanical signal that may play an important role in bone adaptation to its mechanical environment. Previous in vitro studies of osteoblastic cells revealed that the upregulation of cyclooxygenase-2 (COX-2) and c-fos induced by steady fluid flow depends on a change in actin polymerization dynamics and the formation of actin stress fibers. Exposing cells to dynamic oscillatory fluid flow, the temporal flow pattern that results from normal physical activity, is also known to result in increased COX-2 expression and PGE(2) release. The purpose of this study was to determine whether dynamic fluid flow results in changes in actin dynamics similar to steady flow and to determine whether alterations in actin dynamics are required for PGE(2) release. We found that exposure to oscillatory fluid flow did not result in the development of F-actin stress fibers in MC3T3-E1 osteoblastic cells and that inhibition of actin polymerization with cytochalasin D did not inhibit intracellular calcium mobilization or PGE(2) release. In fact, PGE(2) release was increased threefold in the polymerization inhibited cells and this PGE(2) release was dependent on calcium release from the endoplasmic reticulum. This was in contrast to the PGE(2) release that occurs in normal cells, which is independent of calcium flux from endoplasmic reticulum stores. We suggest that this increased PGE(2) release involves a different molecular mechanism perhaps involving increased deformation due to the compromised cytoskeleton.
Malone
Amanda M D
AM
Bone and Joint Rehabilitation R and D Center, Veterans Affairs Medical Center, Palo Alto, CA, USA.
Batra
Nikhil N
NN
Shivaram
Giri
G
Kwon
Ron Y
RY
You
Lidan
L
Kim
Chi Hyun
CH
Rodriguez
Joshua
J
Jair
Kai
K
Jacobs
Christopher R
CR
D016475:3T3 Cells; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001696:Biomechanical Phenomena; D002118:Calcium / Q000378:metabolism; D048430:Cell Shape; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D015232:Dinoprostone / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D040542:Mechanotransduction, Cellular / Q000187:drug effects; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D011673:Pulsatile Flow; D022502:Stress Fibers / Q000378:metabolism; D013314:Stress, Mechanical; D015854:Up-Regulation
D000199:Actins; D015638:Cytochalasin D; D015232:Dinoprostone; D002118:Calcium