Stimulation of protein and DNA synthesis in mouse C2C12 satellite cells: evidence for phospholipase D-dependent and -independent pathways.

Morrison KS; Mackie SC; Palmer RMThompson MG

摘要

In C2C12 myoblasts, 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated a phospholipase D (PLD) to degrade phosphatidylcholine (PC) as measured by the release of choline and an increase in the formation of phosphatidic acid (PA) (or phosphatidylbutanol [PtdBuOH] in the presence of 0.5% butanol). Exogenous PLD also stimulated choline release, PA and PtdBuOH formation. The protein kinase C (PKC) inhibitor, Ro-31-8220, and PKC downregulation significantly inhibited the effects of TPA but Ro-31-8220 had no effect on PLD action. Neither basic Fibroblast Growth Factor (bFGF) or Epidermal Growth Factor (EGF) increased PLD activity. All agonists stimulated protein synthesis during both a 90 min and a 6 hr incubation and increased RNA accretion after 6 hr. The response at 90 min was not inhibited by the transcription inhibitor, actinomycin D. Ro-31-8220 and PKC downregulation significantly inhibited all the effects of TPA. In contrast, Ro-31-8220 significantly inhibited the increase in RNA accretion elicited by PLD but had no effect on the ability of agonists other than TPA to enhance protein synthesis. All agonists also stimulated thymidine incorporation into DNA. The effects of EGF, bFGF, and PLD were rapid and transient whereas that of TPA was delayed and sustained. Ro-31-8220 and PKC downregulation significantly inhibited the response due to TPA. Furthermore, Ro-31-8220 also significantly inhibited the effects elicited by EGF and PLD but not that induced by bFGF. In differentiated myotubes, TPA and PLD, but not bFGF or EGF, again stimulated choline release and PtdBuOH formation. However, all agents failed to stimulate protein synthesis and RNA accretion. The data demonstrate the presence in C2C12 myoblasts, but not differentiated myotubes, of both a PLD-dependent and PLD-independent pathway(s) leading to the stimulation of protein synthesis, RNA accretion, and DNA synthesis.

机译：在C2C12成肌细胞,12-O-tetradecanoylphorbol-13-acetate (TPA)刺激磷脂酶D(骑士)降解磷脂酰胆碱(PC)的释放的胆碱和增加(或形成磷脂酸(PA)phosphatidylbutanol [PtdBuOH]的存在丁醇0.5%)。胆碱释放,PA和PtdBuOH形成。蛋白激酶C (PKC)抑制剂,ro - 31 - 8220,显著差别PKC对这些基因的抑制TPA的影响但ro - 31 - 8220没有影响骑士的行动。因子(bFGF)和表皮生长因子(EGF)骑士活动增加。蛋白质合成在90分钟和6小时孵化和增加RNA后吸积6人力资源。转录抑制剂放线菌素D。显著差别ro - 31 - 8220和PKC对这些抑制TPA的影响。ro - 31 - 8220显著抑制增长RNA吸积引起的骑士,但没有影响受体激动剂TPA以外的能力提高蛋白质合成。刺激胸苷并入DNA。表皮生长因子的影响、bFGF和骑士都迅速和瞬态而TPA误点持续。显著抑制由于TPA的响应。此外,ro - 31 - 8220也显著抑制EGF和骑士,但引发的影响bFGF引起的。肌管,TPA和骑士,但不是bFGF或EGF刺激胆碱释放和PtdBuOH形成。然而,所有代理未能刺激蛋白质合成和RNA吸积。出现在C2C12成肌细胞,但不是差异化的肌管,PLD-dependent和PLD-independent通路(s)导致的刺激蛋白质合成的RNA吸积,和DNA合成。

Stimulation of protein and DNA synthesis in mouse C2C12 satellite cells: evidence for phospholipase D-dependent and -independent pathways.

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