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Am J Physiol Regul Integr Comp Physiol 283: R647-R652, 2002. First published June 6, 2002; doi:10.1152/ajpregu.00121.2002
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Vol. 283, Issue 3, R647-R652, September 2002

IGF-I binding in primary culture of muscle cells of rainbow trout: changes during in vitro development

Juan Castillo1, Pierre-Yves Le Bail2, Gilles Paboeuf2, Isabel Navarro1, Claudine Weil2, Benoit Fauconneau2, and Joaquim Gutiérrez1

1 Departament de Fisiologia, Facultat de Biologia, Universitat de Barcelona, E-08028 Barcelona, Spain; and 2 Institut National de la Recherche Agronomique-Station Commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement, Campus de Beaulieu, 35042 Cedex Rennes, France

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 h 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 and 0.95 ± 0.05 nM, respectively). IGF binding proteins did not interfere significantly because displacement of labeled IGF-I by either cold trout recombinant IGF-I or Des (1-3) IGF-I resulted in similar curves. Insulin did not displace labeled IGF-I even at very high concentrations (>1 µM), 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 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.

insulin-like growth factor-I; receptor


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