Mechanisms contributing to fluid-flow-induced Ca2+ mobilization in articular chondrocytes.
We previously showed that fluid flow, which chondrocytes experience in vivo and which results in a variety of morphological and metabolic changes in cultured articular chondrocytes, can also stimulate a rise in intracellular calcium concentration ([Ca2+]i). However, the mechanism by which Ca2+ is mobilized in response to flow is unclear. In this study, we investigated the roles of intracellular Ca2+ stores, G-proteins, and extracellular ATP in the flow-induced Ca2+ response in bovine articular chondrocytes (BAC). Cells loaded with the Ca2+ sensitive dye Fura-2 were exposed to steady flow at 34 ml/min (37 dynes/cm2) in a parallel plate flow chamber. Whereas ryanodine and caffeine had no effect, both neomycin and thapsigargin significantly decreased the Ca2+(i) response to flow, suggesting a role for Ca2+ store release, possibly through an inositol 1,4,5-trisphosphate (IP3)-dependent mechanism. Twenty-four-hour treatment with pertussis toxin also significantly decreased the response, suggesting that the mechanism may be G-protein regulated. In addition, ATP release by chondrocytes does not appear to mediate the flow-induced Ca2+ response because suramin, a P2 purinergic blocker, had no effect. These results suggest that BAC respond rapidly to changes in their mechanical environment, such as increased fluid flow, by a mechanism that involves IP3 stimulated Ca2+(i) release and G-protein activation.
Yellowley
C E
CE
Department of Orthopaedics and Rehabilitation, The Pennsylvania State University College of Medicine, Hershey 17033, USA. cyellowl@ortho.hmc.psu.edu
Jacobs
C R
CR
Donahue
H J
HJ
D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D001826:Body Fluids / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D002358:Cartilage, Articular / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000502:physiology; D007425:Intracellular Membranes / Q000378:metabolism
D000255:Adenosine Triphosphate; D019204:GTP-Binding Proteins; D002118:Calcium