IGF-I binding in primary culture of muscle cells of rainbow trout: changes during in vitro development - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Access content directly
Journal Articles AJP - Regulatory, Integrative and Comparative Physiology Year : 2002

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

Abstract

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 onday 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.

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hal-02677784 , version 1 (31-05-2020)

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Juan Castillo, Pierre-Yves Le Bail, Gilles Paboeuf, Isabel Navarro, Claudine Weil, et al.. IGF-I binding in primary culture of muscle cells of rainbow trout: changes during in vitro development. AJP - Regulatory, Integrative and Comparative Physiology, 2002, 283 (3), pp.R647-R652. ⟨10.1152/ajpregu.00121.2002⟩. ⟨hal-02677784⟩

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