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目的关于蛋白激酶C(PKC)在神经元突起生长和神经再生中的作用,目前仍存有争议。本研究主要观察PKC对离体培养的脊髓神经元生长的调节作用,旨在阐明PKC对突起生长的调节作用。方法分离纯化胎龄14天(E14)的SD胎鼠的脊髓前角神经元,进行原代培养,并检测不同时相点膜/浆PKC活性(m/c-PKCactivity)的比值。结果神经元培养3-11d期间,神经元内m/c-PKC比值以及PKC-βII在突起中的表达水平均与突起生长呈显著相关关系(r=0.95,P<0.01;r=0.73,P<0.01)。此外,PKC激动剂PMA能显著提高m/c-PKC比值,且与神经突起的生长一致(r=0.99,P<0.01)。而PKC抑制剂GF109203X则能显著抑制突起生长,且不被PMA作用所逆转。结论PKC的活性在脊髓神经元突起生长调节中具有重要作用,其中βII亚型可能扮演重要角色。
Purpose The role of protein kinase C (PKC) in neurite outgrowth and neurogenesis remains controversial. This study mainly observed the regulatory effect of PKC on the growth of spinal cord neurons in vitro and aimed to elucidate the regulatory effect of PKC on neurite outgrowth. Methods The spinal cord anterior horn neurons were isolated and purified from embryonic day 14 (E14) embryos and cultured in primary culture. The ratio of membrane / plasma PKC activity (m / c-PKC activity) at different time points was measured. Results The neuronal m / c-PKC ratio and the expression level of PKC-βII in the neurites were significantly correlated with the neurite outgrowth (r = 0.95, P <0.01; r = 0.73, P <0.01). In addition, PKC agonist PMA significantly increased m / c-PKC ratio and was consistent with neurite growth (r = 0.99, P <0.01). The PKC inhibitor GF109203X can significantly inhibit the growth of the protrusion, and is not reversed by PMA. Conclusion The activity of PKC plays an important role in the regulation of neurite outgrowth in spinal cord. The βII subtype may play an important role.