Centre for Discovery Brain Science, University of Edinburgh, Hugh Robson Building, Edinburgh EH8 9XD, UK; Euan McDonald Centre for Motor Neuron Disease Research, University of Edinburgh, UK; The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, EH25 9RG, UK.
Euan McDonald Centre for Motor Neuron Disease Research, University of Edinburgh, UK; Centre for Cognitive and Neural Systems, University of Edinburgh, 1 George Square, Edinburgh EH8 9JZ, UK.
Neurobiol Dis. 2019 Oct;130:104496. doi: 10.1016/j.nbd.2019.104496. Epub 2019 Jun 6.
Neurodegenerative and neuromuscular disorders can manifest throughout the lifespan of an individual, from infant to elderly individuals. Axonal and synaptic degeneration are early and critical elements of nearly all human neurodegenerative diseases and neural injury, however the molecular mechanisms which regulate this process are yet to be fully elucidated. Furthermore, how the molecular mechanisms governing degeneration are impacted by the age of the individual is poorly understood. Interestingly, in mice which are under 3 weeks of age, the degeneration of axons and synapses following hypoxic or traumatic injury is significantly slower. This process, known as Wallerian degeneration (WD), is a molecularly and morphologically distinct subtype of neurodegeneration by which axons and synapses undergo distinct fragmentation and death following a range of stimuli. In this study, we first use an ex-vivo model of axon injury to confirm the significant delay in WD in neonatal mice. We apply tandem mass-tagging quantitative proteomics to profile both nerve and muscle between P12 and P24 inclusive. Application of unbiased in silico workflows to relevant protein identifications highlights a steady elevation in oxidative phosphorylation cascades corresponding to the accelerated degeneration rate. We demonstrate that inhibition of Complex I prevents the axotomy-induced rise in reactive oxygen species and protects axons following injury. Furthermore, we reveal that pharmacological activation of oxidative phosphorylation significantly accelerates degeneration at the neuromuscular junction in neonatal mice. In summary, we reveal dramatic changes in the neuromuscular proteome during post-natal maturation of the neuromuscular system, and demonstrate that endogenous dynamics in mitochondrial bioenergetics during this time window have a functional impact upon regulating the stability of the neuromuscular system.
神经退行性和神经肌肉疾病可以在个体的整个生命周期中表现出来,从婴儿到老年人。轴突和突触退化是几乎所有人类神经退行性疾病和神经损伤的早期和关键因素,然而,调节这一过程的分子机制尚未完全阐明。此外,调节退化的分子机制如何受到个体年龄的影响还知之甚少。有趣的是,在年龄小于 3 周的小鼠中,缺氧或创伤性损伤后轴突和突触的退化明显较慢。这个过程被称为沃勒变性(Wallerian degeneration,WD),是一种分子和形态上不同的神经退行性变亚型,其中轴突和突触在受到一系列刺激后经历明显的碎片化和死亡。在这项研究中,我们首先使用轴突损伤的离体模型来确认新生小鼠 WD 显著延迟。我们应用串联质量标记定量蛋白质组学技术,在 P12 至 P24 之间对神经和肌肉进行全面分析。对相关蛋白质鉴定的无偏倚计算流程应用突出显示氧化磷酸化级联的稳定升高,对应于加速的退化率。我们证明,抑制复合物 I 可以防止轴突切断诱导的活性氧增加,并在损伤后保护轴突。此外,我们揭示了氧化磷酸化的药理学激活在新生小鼠的神经肌肉接头显著加速退化。总之,我们揭示了神经肌肉系统出生后成熟过程中神经肌肉蛋白质组的巨大变化,并证明了在此时间窗口内线粒体生物能量学的内源性动态对调节神经肌肉系统的稳定性具有功能影响。