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Articles in PresS, published online ahead of print June 6, 2002
Am J Physiol Regu Physiol, 10.1152/ajpregu.00121.2002
Submitted on February 22, 2002
Accepted on May 30, 2002
1 Departamento de Fisiologia, Universitat de Barcelona, Barcelona, Spain
2 Institut Nationale de la recherche agronomique (INRA), Rennes, France
* To whom correspondence should be addressed. E-mail: joaquim{at}porthos.bio.ub.es.
To characterize and study the variations of IGF-I binding during the development of trout muscle cells, in vitro experiments were conducted using myocyte cultures and IGF-I binding assays were performed in three stages of cell development: mononuclear cells (day 1), small myotubes (day 4) and large myotubes (day 10). Binding experiments were done by incubating cells with IGF-I for 12 hours at 4°C. Specific IGF-I binding increased with the concentration of labeled IGF-I and reached a plateau at 32 pM. The displacement of cold human and trout IGF-I showed a very similar curve (EC50=1.19 ± 0.05 nM and 0.95 ± 0.05 nM, respectively). IGF binding proteins (IGFBP) did not interfere significantly since displacement of labeled IGF-I by either cold rtIGF-I or Des (1-3) IGF-I resulted in similar curves. Insulin did not displace labeled IGF-I even at very high concentrations (>1µM), which indicates the specificity of IGF-I binding. The amount of receptor (R0) increased from 253 ± 51 fmol/mg DNA on day 1 to 766 ± 107 fmol/mg DNA on day 10. However, the affinity (Kd) of IGF-I receptors did not change significantly during this development (from 1.29 ± 0.19 nM to 0.79 ± 0.13 nM). On the basis of our results, we conclude that rainbow trout muscle cells in culture express specific IGF-I receptors, which increase their number with development from mononuclear cells to large myotubes.
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