Research Article
Effects of interleukin-1 on calcium signaling and the increase of filamentous actin in isolated and in situ articular chondrocytes
Scott Pritchard,
Farshid Guilak,
Scott Pritchard
Duke University Medical Center, Durham, North Carolina
Search for more papers by this authorCorresponding Author
Farshid Guilak
Duke University Medical Center, Durham, North Carolina
Orthopaedic Research Laboratories, Duke University Medical Center, 375 Medical Sciences Research Building, Research Drive, Box 3093, Durham, NC 27710Search for more papers by this authorScott Pritchard,
Farshid Guilak,
Scott Pritchard
Duke University Medical Center, Durham, North Carolina
Search for more papers by this authorCorresponding Author
Farshid Guilak
Duke University Medical Center, Durham, North Carolina
Orthopaedic Research Laboratories, Duke University Medical Center, 375 Medical Sciences Research Building, Research Drive, Box 3093, Durham, NC 27710Search for more papers by this authorAbstract
Objective
To determine whether interleukin-1 (IL-1) initiates transient changes in the intracellular concentration of [Ca2+]i and the organization of filamentous actin (F-actin) in articular chondrocytes.
Methods
Articular chondrocytes within cartilage explants and enzymatically isolated chondrocytes were loaded with Ca2+-sensitive fluorescence indicators, and [Ca2+]i was measured using confocal fluorescence ratio imaging during exposure to 10 ng/ml IL-1α. Inhibitors of Ca2+ mobilization (Ca2+-free medium, thapsigargin [inhibitor of Ca-ATPases], U73122 [inhibitor of phospholipase C], and pertussis toxin [inhibitor of G proteins]) were used to determine the mechanisms of increased [Ca2+]i. Cellular F-actin was quantified using fluorescently labeled phalloidin. Toxin B was used to determine the role of the Rho family of small GTPases in F-actin reorganization.
Results
In isolated cells on glass and in in situ chondrocytes within explants, exposure to IL-1 induced a transient peak in [Ca2+]i that was generally followed by a series of decaying oscillations. Thapsigargin, U73122, and pertussis toxin inhibited the percentage of cells responding to IL-1. IL-1 increased F-actin content in chondrocytes in a manner that was inhibited by toxin B.
Conclusion
Both isolated and in situ chondrocytes respond to IL-1 with transient increases in [Ca2+]i via intracellular Ca2+ release mediated by the phospholipase C and inositol trisphosphate pathways. The influx of Ca2+ from the extracellular space and the activation of G protein–coupled receptors also appear to contribute to these mechanisms. These findings suggest that Ca2+ mobilization may be one of the first signaling events in the response of chondrocytes to IL-1.
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