A transient increase of snoN transcript by growth arrest upon serum deprivation and cell-to-cell contact

To analyze the possible involvement of c-ski and c-sno during the course of in vitro myogenesis, expression of their transcripts during differentiation of a murine muscle cell line (C2C12) was monitored by competitive reverse transcription-polymerase chain reaction (RT-PCR). The transcripts of c-snoN were temporarily increased 25-fold above basal level at 12 h prior to the onset of transcription of muscle-specific gene, e.g. myogenin and muscle creatine kinase, whereas c-ski was expressed invariably. The transient increase of c-snoN was blocked when myogenesis was interrupted by the presence of fetal calf serum in culture medium, probably due to growth factors being included; basic fibroblast growth factor (b-FGF) blocked the transient increase whereas epidermal growth factor (EGF) did not, consistent with the inhibitory effect of b-FGF and no effect of EGF on myotube formation of C2C12. In fibroblastic C3H10T1/2 cells, snoN exhibited a similar transient increase of transcript when growth arrested under the same conditions as for in vitro myogenesis, indicating that the expression of snoN is not sufficient to induce the onset of muscle differentiation and an unknown factor involved in myogenic cells is necessary. The transient increase of snoN transcript may represent a common entrance step of cells into the G0 phase where muscle differentiation is substantiated, considering that it was observed upon growth arrest of fibroblastic C3H10T1/2 cells and prior to the elevation of MCK in C2C12 but undetected when entry into G0 was blocked by b-FGF.