GENERATION OF GROWTH-HORMONE BINDING-PROTEIN BY AVIAN GROWTH-PLATE CHONDROCYTES IS DEPENDENT ON CELL-DIFFERENTIATION

Growth hormone receptor (GH-R) gene expression was evaluated in avian growth-plates in situ and in cultured chondrocytes. In the epiphyseal growth-plate, chondrocytes at different stages of differentiation loca ted at the proliferative and upper hypertrophic zones express the GH-R gene. In culture, addition of ascorbic acid facilitated chondrocyte d ifferentiation as evaluated by decrease in collagen type II gene expre ssion and increase in alkaline phosphatase activity and osteopontin ge ne expression. Both the ascorbic acid-treated and untreated chondrocyt es expressed the gene coding for the chicken growth-hormone receptor ( cGH-R), but only the undifferentiated cells were capable of binding th e hormone. This reduction in GH-binding resulted in alteration in GH-d ependent regulation of the GH-R gene expression: only the undifferenti ated chondrocytes responded to chicken GH (cGH) by down-regulation of the cGH-R gene expression. Chondrocyte differentiation induced by eith er ascorbic acid or retinoic acid was associated with the appearance o f two growth hormone binding-proteins (GHBPs) in the culture medium wi th estimated MWs of 32 and 70 kDa, respectively. These GHBPs differ in their MW from the major GHBP found in chicken plasma. Chondrocyte GHB Ps specifically bind [I-125]CGH, which can be displaced by an excess o f unlabeled cGH. The differentiation-dependent increase in the 70 kDa GHBP was observed also using specific chicken GHBP antiserum. Our data suggest that the reduction of the differentiated chondrocytes respons e to GH is due to differentiation-dependent loss of the extracellular domain of the GH-R, resulting in a lack of functional receptors on the cell surface and generation of GHBP. (C) 1997 Elsevier Science Irelan d Ltd.