Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage.
OBJECTIVE
Connexin (Cx) 43 hemichannels play a role in mechanotransduction. This study was undertaken in order to determine if Cx43 hemichannels were activated in rat temporomandibular joint (TMJ) chondrocytes under mechanical stimulation.


METHODS
Sprague-Dawley rats were stimulated dental-mechanically. Cx43 expression in rat TMJ cartilage was determined with immunohistochemistry and real-time PCR, and Cx43 hemichannel opening was evaluated by the extra- and intracellular levels of prostaglandin E2 (PGE2). Both primary rat chondrocytes and ATDC5 cells were treated with fluid flow shear stress (FFSS) to induce hemichannel opening. The Cx43 expression level was then determined by real-time PCR or Western blotting, and the extent of Cx43 hemichannel opening was evaluated by measuring both PGE2 release and cellular dye uptake.


RESULTS
Cx43 expression and intra- and extracellular PGE2 levels were increased in mechanically-stimulated rat TMJ cartilage compared to the unstimulated control. The FFSS treatment increased Cx43 expression and induced Cx43 hemichannel opening in primary rat chondrocytes and ATDC5 cells indicated by enhanced PGE2 release and dye uptake. Furthermore, the Cx43 hemichannel opening could be blocked by the addition of 18β-glycyrrhetinic acid, a Cx channel inhibitor, Cx43-targeting siRNA, or by withdrawal of FFSS stimulation. The migration of cytosolic Cx43 protein to the plasma membrane in ATDC5 cells was still significant after 8 h post 2-h FFSS treatment, and the Cx43 protein level was still high at 48 h, which returned to control levels at 72 h after treatment.


CONCLUSIONS
Cx43 hemichannels are activated and mediate small molecule exchange between TMJ chondrocytes and matrix under mechanical stimulation.
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China.
J
J
Zhang
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China.
H Y
HY
Zhang
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China.
M
M
Zhang
College of Life Science, Shaanxi Normal University, Xi'an, 710062, China.
Z Y
ZY
Qiu
Institute of Orthopaedics, Xijing Hospital, Fourth Military Medical University, 15 Changlexi Road, Xi'an, 710032, China.
Y P
YP
Wu
Department of Biochemistry, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
D A
DA
Callaway
Department of Biochemistry, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
J X
JX
Jiang
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China.
L
L
Lu
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China.
L
L
Jing
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China.
T
T
Yang
State Key Laboratory of Military Stomatology, Department of Oral Anatomy and Physiology, School of Stomatology, Fourth Military Medical University, 145 Changlexi Road, Xi'an, 710032, China. Electronic address: mqwang@fmmu.edu.cn.
M Q
MQ
Wang
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism / Q000648:ultrastructure; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D015232:Dinoprostone / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D007150:Immunohistochemistry; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D018613:Microscopy, Confocal; D034741:RNA, Small Interfering / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012016:Reference Values; D012680:Sensitivity and Specificity; D013314:Stress, Mechanical; D013704:Temporomandibular Joint / Q000378:metabolism* / Q000473:pathology
D018031:Connexin 43; D034741:RNA, Small Interfering; D015232:Dinoprostone
Animal model; Chondrocyte; Connexin43; Hemichannel; Mechanical loading; Osteoarthritis