Stimulation of regulatory volume increase (RVI) in avian articular chondrocytes by gadolinium chloride

Abstract

Chondrocytes, the resident cell-type of articular cartilage, are responsible for the regulation of the extracellular matrix (ECM) in response to their physico-chemical environment. Due to the nature of cartilage loading, chondrocytes are exposed to constant changes in extracellular osmolality with a gradual increase throughout the day. As an increase in osmolality attenuates matrix synthesis, we have studied cell volume regulation (regulatory volume increase (RVI)) after hypertonic challenge and the regulation of RVI by the actin cytoskeleton. Using freshly isolated avian articular chondrocytes, changes in actin organisation were studied by confocal laser scanning microscopy following a 43% increase in extracellular osmolality. Using calcein-loading chondrocytes, the capacity for RVI was determined and the rate of volume recovery (t1/2) mathematically extrapolated. Following an increase in extracellular osmolality there was a significant increase (p < 0.05) in cortical actin, inhibited by the removal of extracellular calcium EGTA or by the addition of 100 micromol.L-1 gadolinium chloride. Most cells exhibited slow RVI (t1/2 = 55.5 +/- 5.5 min), whereby inhibition of actin polymerisation by gadolinium chloride or the removal of extracellular calcium significantly increased the rate of volume recovery via a bumetanide-sensitive pathway (t1/2 of 29.6 +/- 6.5 min and 13.8 +/- 3.1 min, respectively). These data suggest the Na+-K+-2Cl(-) (NKCC) co-transporter regulated by the actin cytoskeleton is involved in avian chondrocyte RVI.