Growth hormone stimulates the secretion of insulin-like growth factor binding protein-2 (IGFBP-2) by monolayer cultures of sheep costal growth plate chondrocytes.
Using monolayer cultures of costal chondrocytes established from four week old Clun Forest lambs, we have demonstrated that, under serum free conditions the cells release three IGFBPs (32, 29 and 21 kDa) into the medium. The most abundant of these--the 32 kDa BP--was shown to be IGFBP-2 by Western blotting. Furthermore we demonstrate that the levels of IGFBP-2 in conditioned medium are acutely increased (6, 12 and 24 h time points) following treatment of cells with bovine GH (1-100 ng/ml). In a parallel set of experiments, using ovine fibroblasts (derived from dermis) we show that IGFBPs of Mr 32, 29 and 21 kDa are also secreted by this cell type. However the relative abundance of these BPs differed from that seen in the chondrocyte cultures, with the 21 kDa species now the most abundant. In addition, prolonged exposure of autoradiographs indicated that fibroblasts secreted a higher Mr IGFBP (most probably ovine BP-3) that was not detected in any of our chondrocyte cultures. Most significant however was the demonstration that bGH did not dramatically affect the levels of IGFBPs in fibroblast cell cultures. We conclude that GH stimulates BP-2 production from chondrocytes and this is a cell-type specific effect in as much as it is not replicated in cultures of dermal fibroblasts.
Borromeo
V
V
Institute of Veterinary Physiology, University of Milan, Italy.
Bramani
S
S
Holder
A T
AT
Carter
C
C
Secchi
C
C
Beattie
J
J
D000818:Animals; D015153:Blotting, Western; D002356:Cartilage / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D013006:Growth Hormone / Q000378:metabolism*; D006132:Growth Plate / Q000166:cytology / Q000378:metabolism*; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000378:metabolism*; D008297:Male; D011994:Recombinant Proteins / Q000378:metabolism; D012272:Ribs; D012756:Sheep
D018971:Insulin-Like Growth Factor Binding Protein 2; D011994:Recombinant Proteins; D013006:Growth Hormone