Department of Biological Sciences, The State University of New York at Buffalo, Buffalo, NY, 14260, US.
The State University of New York at Buffalo, 109 Cooke Hall, North/Amherst Campus, Buffalo, NY, 14260, US.
Apoptosis. 2019 Apr;24(3-4):341-358. doi: 10.1007/s10495-019-01520-4.
High levels of oxidative stress is detected in neurons affected by many neurodegenerative diseases, including huntington's disease. Many of these diseases also show neuronal cell death and axonal transport defects. While nuclear inclusions/accumulations likely cause cell death, we previously showed that cytoplasmic axonal accumulations can also contribute to neuronal death. However, the cellular mechanisms responsible for activating cell death is unclear. One possibility is that perturbations in normal axonal transport alter the function of the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT)-pathway, a signal transduction pathway that promotes survival/growth in response to extracellular signals. To test this proposal in vivo, we expressed active PI3K in the context of pathogenic huntingtin (HTT-138Q) in Drosophila larval nerves, which show axonal transport defects and neuronal cell death. We found that excess expression of active P13K significantly suppressed HTT-138Q-mediated neuronal cell death, but had no effect on HTT-138Q-mediated axonal transport defects. Expression of active PI3K also rescued Paraquat-mediated cell death. Further, increased levels of pSer9 (inactive) glycogen synthase kinase 3β was seen in HTT-138Q-mediated larval brains, and in dynein loss of function mutants, indicating the modulation of the pro-survival pathway. Intriguingly, proteins in the PI3K/AKT-pathway showed functional interactions with motor proteins. Taken together our observations suggest that proper axonal transport is likely essential for the normal function of the pro-survival PI3K/AKT-signaling pathway and for neuronal survival in vivo. These results have important implications for targeting therapeutics to early insults during neurodegeneration and death.
高水平的氧化应激在许多神经退行性疾病(包括亨廷顿病)影响的神经元中被检测到。这些疾病也表现出神经元细胞死亡和轴突运输缺陷。虽然核内包涵体/积聚可能导致细胞死亡,但我们之前已经表明,细胞质轴突积聚也可能导致神经元死亡。然而,负责激活细胞死亡的细胞机制尚不清楚。一种可能性是,正常轴突运输的干扰改变了磷酸肌醇 3-激酶(PI3K)-蛋白激酶 B(AKT)-信号通路的功能,该信号通路可响应细胞外信号促进存活/生长。为了在体内验证这一假设,我们在果蝇幼虫神经中表达了致病亨廷顿蛋白(HTT-138Q)背景下的活性 PI3K,这些神经表现出轴突运输缺陷和神经元细胞死亡。我们发现,过量表达活性 P13K 显著抑制了 HTT-138Q 介导的神经元细胞死亡,但对 HTT-138Q 介导的轴突运输缺陷没有影响。活性 PI3K 的表达也挽救了百草枯介导的细胞死亡。此外,在 HTT-138Q 介导的幼虫大脑中以及在动力蛋白功能丧失突变体中,观察到 pSer9(失活)糖原合酶激酶 3β的水平增加,表明生存途径的调节。有趣的是,PI3K/AKT 信号通路中的蛋白质与运动蛋白表现出功能相互作用。总之,我们的观察结果表明,适当的轴突运输可能是生存的 PI3K/AKT 信号通路的正常功能和体内神经元存活所必需的。这些结果对于在神经退行性变和死亡过程中针对早期损伤靶向治疗具有重要意义。