Components of signaling pathways for insulin and insulin-like growth factor-I in muscle myoblasts and myotubes.
To survey and compare the signaling pathways from the insulin and insulin-like growth factor-I (IGF-I) receptors in undifferentiated and differentiated muscle cells, we examined the phosphotyrosine (Ptyr)-containing polypeptides elicited in L6 and Sol8 myoblasts and myotubes by the combination of insulin and IGF-I. These polypeptides were detected by immunoblotting with antibodies against Ptyr. In the L6 myoblasts and myotubes and the Sol8 myoblasts, Ptyr polypeptides of approximately 240, 175, 115, 100, 41, and 37 kilodaltons (kDa) appeared in response to insulin-IGF-I. With the Sol8 myotubes, the 240-, 175-, and 37-kDa Ptyr polypeptides were detected in basal cells, and only the Ptyr content of the 175-kDa one increased in response to insulin-IGF-I. The polypeptides of 175, 41, and 37 kDa were tentatively identified as the insulin receptor substrate 1 (IRS1) and extracellular signal-regulated kinases 1 and 2 (ERK1 and -2), respectively, by immunoblotting with antibodies specific for these proteins, and the 115- and 100-kDa polypeptides are probably the beta-subunits of the insulin and IGF-I receptors. The amounts of IRS1, ERK1, and ERK2 were roughly the same in the L6 and Sol8 myoblasts and myotubes. Thus, differentiation of the myoblasts to myotubes was not accompanied by the detectable appearance of new insulin-IGF-I-elicited Ptyr polypeptides or marked changes in the amounts of known participants in their signaling pathways.
Lamphere
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Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755-3844.
Lienhard
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GE
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002478:Cells, Cultured; D015151:Immunoblotting; D007328:Insulin / Q000494:pharmacology / Q000502:physiology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology / Q000502:physiology*; D051379:Mice; D009132:Muscles / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D010766:Phosphorylation; D010770:Phosphotransferases / Q000502:physiology; D019000:Phosphotyrosine; D051381:Rats; D011972:Receptor, Insulin / Q000032:analysis / Q000502:physiology; D017451:Receptors, Somatomedin / Q000032:analysis / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D007328:Insulin; D017451:Receptors, Somatomedin; D019000:Phosphotyrosine; D014443:Tyrosine; D007334:Insulin-Like Growth Factor I; D010770:Phosphotransferases; D011972:Receptor, Insulin