دورية أكاديمية
IGF-I binding and receptor signal transduction in primary cell culture of muscle cells of gilthead sea bream: changes throughout in vitro development.
العنوان: | IGF-I binding and receptor signal transduction in primary cell culture of muscle cells of gilthead sea bream: changes throughout in vitro development. |
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المؤلفون: | Montserrat N; Departament de Fisiologia, Facultat de Biologia, Universitat de Barcelona, 08028 Barcelona, Spain. feliz119@gmail.com, Sánchez-Gurmaches J, García de la Serrana D, Navarro MI, Gutiérrez J |
المصدر: | Cell and tissue research [Cell Tissue Res] 2007 Dec; Vol. 330 (3), pp. 503-13. Date of Electronic Publication: 2007 Oct 17. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 0417625 Publication Model: Print-Electronic Cited Medium: Print ISSN: 0302-766X (Print) Linking ISSN: 0302766X NLM ISO Abbreviation: Cell Tissue Res Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: Berlin, New York, Springer-Verlag. |
مواضيع طبية MeSH: | Signal Transduction*, Fish Proteins/*metabolism , Insulin-Like Growth Factor I/*metabolism , Muscle Fibers, Skeletal/*cytology , Muscle Fibers, Skeletal/*metabolism , Receptor, IGF Type 1/*metabolism , Sea Bream/*growth & development, Animals ; Binding Sites ; Cell Culture Techniques ; Cell Differentiation ; Flow Cytometry ; Mitogen-Activated Protein Kinases/metabolism ; Phosphatidylinositol 3-Kinases/metabolism ; Proto-Oncogene Proteins c-akt/metabolism ; Sea Bream/metabolism |
مستخلص: | We examined the possibility of culturing muscle cells of gilthead sea bream in vitro and assessed variations in insulin-like growth factor-I (IGF-I) binding during myocyte development. The viability of the cell culture was determined by fluorescence-activated cell-sorting analysis, which showed that the percentage of dead cells decreased with cell differentiation. The intracellular reduction of MTT into formazan pigment was preferentially carried out as cells differentiated (from day 4) indicating an increase in metabolic activity. IGF-I-binding assays demonstrated that the number of receptors increased from 190 +/- 0.09 fmol/mg protein in myocytes at day 5 to 360 +/- 0.09 fmol/mg protein in myotubes at day 12. The affinity of IGF-I receptors did not change significantly during cell development (from 0.89 +/- 0.09 to 0.98 +/- 0.09 nM). The activation of various kinase (ERK 1/2 MAPK and Akt/PKB) proteins by IGFs and insulin was studied by means of Western blot analysis. Levels of MAPK-P increased after IGF and insulin treatment during the first stages of cell culture, with a low response being observed at day 15, whereas IGFs displayed a stimulatory effect on Akt-P throughout the cell culture period, even on day 15. This study thus shows that (1) gilthead sea bream myocytes can be cultured, (2) they express functional IGF-I receptors that increase in number as they differentiate in vitro; (3) IGF signalling transduction through IGF-I receptors stimulates the MAPK and Akt pathways, depending on the development stage of the muscle cell culture. |
المشرفين على المادة: | 0 (Fish Proteins) 67763-96-6 (Insulin-Like Growth Factor I) EC 2.7.1.- (Phosphatidylinositol 3-Kinases) EC 2.7.10.1 (Receptor, IGF Type 1) EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) EC 2.7.11.24 (Mitogen-Activated Protein Kinases) |
تواريخ الأحداث: | Date Created: 20071018 Date Completed: 20080411 Latest Revision: 20101118 |
رمز التحديث: | 20240628 |
DOI: | 10.1007/s00441-007-0507-2 |
PMID: | 17940802 |
قاعدة البيانات: | MEDLINE |
تدمد: | 0302-766X |
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DOI: | 10.1007/s00441-007-0507-2 |