Chicken serum transferrin duplicates the myotrophic effects of sciatin on cultured muscle cells.
Sciatin, a glycoprotein purified from chicken sciatic nerves, has been shown to have trophic effects on chicken skeletal muscle cells in culture. Since we recently observed pronounced structural similarities between sciatin and chicken serum transferrin [Markelonis et al, 1982a], we decided to investigate the muscle growth-promoting activity of transferrin on cultured muscle cells. Serum transferrin was isolated by the same protocol used to purify sciatin, viz., affinity chromatography on concanavalin A-agarose followed by ion-exchange chromatography on DEAE cellulose. The serum protein recovered by this purification scheme was indistinguishable immunologically from sciatin as evidenced by a positive precipitin reaction against goat anti-sciatin serum on double immunodiffusion in agar. Purified serum transferrin had myotrophic effects identical to those of sciatin when added to skeletal muscle cells in vitro. For example, even when chicken embryo extract--a constituent normally required for chicken muscle cell differentiation in vitro--was omitted from culture medium, either serum transferrin or sciatin promoted myogenesis in culture as measured by a stimulation of the fusion index. Furthermore, both proteins caused a significant increase in the level of protein synthesis, the number of acetylcholine receptors and the activity of acetylcholinesterase in treated muscle cultures. By contrast, commercially obtained ovotransferrin (conalbumin) or FeSO4 (100 microM) were unable to fully support myogenesis of skeletal muscle in vitro if embryo extract was omitted from the culture medium. From these data, we conclude that the neuronal myotrophic protein sciatin is both structurally and biologically related to serum transferrin. Furthermore, we suggest that sciatin may represent a neuronal form of this iron-transport protein.
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002642:Chick Embryo; D002645:Chickens / Q000097:blood; D046508:Culture Techniques; D007501:Iron / Q000502:physiology; D009124:Muscle Proteins / Q000096:biosynthesis; D009132:Muscles / Q000187:drug effects*; D009419:Nerve Tissue Proteins / Q000494:pharmacology*; D011950:Receptors, Cholinergic / Q000032:analysis; D014168:Transferrin / Q000097:blood / Q000494:pharmacology*
D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D011950:Receptors, Cholinergic; D014168:Transferrin; C026913:sciatin; D007501:Iron