Modulation of type X collagen gene expression by calcium β-glycerophosphate and levamisole: implications for a possible role for type X collagen in endochondral bone formation

Abstract

Chondrocytes from the cephalic region of 18-day chick embryo sterna were cultured within type I collagen gels in the presence of increasing concentrations of calcium beta-glycerophosphate (Ca beta GP) (2.5 mM, 5 mM and 10 mM) or 2 mM levamisole. Addition of Ca beta GP produced an increase in type X collagen synthesis and when 10 mM Ca beta GP was used, mineral deposits were observed in the matrix elaborated by the chondrocytes. Measurement of mRNA levels of type II and type X collagen microgram-1 total RNA revealed that the level of alpha 1(II) mRNA decreased whereas the level of alpha 1(X) mRNA remained relatively constant in the presence of Ca beta GP. Thus, Ca beta GP appears to increase type X collagen synthesis by enhancing the rate of collagen type X mRNA translation. In levamisole-treated cultures the synthesis of type X collagen and its deposition within the matrix were inhibited. Measurement of mRNA levels micrograms-1 total RNA revealed that the level of alpha 1(II) mRNA increased whereas the level of alpha 1(X) mRNA was markedly decreased in the presence of levamisole. Consequently, it must be concluded that levamisole inhibits type X collagen synthesis by inhibiting type X collagen gene transcription. These results add support for type X collagen having an important role in the mineralization process and have demonstrated that type X collagen synthesis is probably regulated at the mRNA translational stage during chondrocyte hypertrophy.