Molecular PathoBiology, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
PLoS Pathog. 2012;8(11):e1003002. doi: 10.1371/journal.ppat.1003002. Epub 2012 Nov 8.
Prion diseases typically have long pre-clinical incubation periods during which time the infectious prion particle and infectivity steadily propagate in the brain. Abnormal neuritic sprouting and synaptic deficits are apparent during pre-clinical disease, however, gross neuronal loss is not detected until the onset of the clinical phase. The molecular events that accompany early neuronal damage and ultimately conclude with neuronal death remain obscure. In this study, we used laser capture microdissection to isolate hippocampal CA1 neurons and determined their pre-clinical transcriptional response during infection. We found that gene expression within these neurons is dynamic and characterized by distinct phases of activity. We found that a major cluster of genes is altered during pre-clinical disease after which expression either returns to basal levels, or alternatively undergoes a direct reversal during clinical disease. Strikingly, we show that this cluster contains a signature highly reminiscent of synaptic N-methyl-D-aspartic acid (NMDA) receptor signaling and the activation of neuroprotective pathways. Additionally, genes involved in neuronal projection and dendrite development were also altered throughout the disease, culminating in a general decline of gene expression for synaptic proteins. Similarly, deregulated miRNAs such as miR-132-3p, miR-124a-3p, miR-16-5p, miR-26a-5p, miR-29a-3p and miR-140-5p follow concomitant patterns of expression. This is the first in depth genomic study describing the pre-clinical response of hippocampal neurons to early prion replication. Our findings suggest that prion replication results in the persistent stimulation of a programmed response that is mediated, at least in part, by synaptic NMDA receptor activity that initially promotes cell survival and neurite remodelling. However, this response is terminated prior to the onset of clinical symptoms in the infected hippocampus, seemingly pointing to a critical juncture in the disease. Manipulation of these early neuroprotective pathways may redress the balance between degeneration and survival, providing a potential inroad for treatment.
朊病毒病通常具有较长的临床前潜伏期,在此期间,感染性朊病毒颗粒和感染性稳步在大脑中传播。在临床前疾病期间,异常的神经突发芽和突触缺陷是明显的,然而,直到临床阶段开始才检测到大量神经元丢失。伴随早期神经元损伤并最终导致神经元死亡的分子事件仍然不清楚。在这项研究中,我们使用激光捕获显微切割分离海马 CA1 神经元,并确定了它们在感染期间的临床前转录反应。我们发现,这些神经元内的基因表达是动态的,并具有不同的活性阶段。我们发现,一大类基因在临床前疾病期间发生改变,然后表达要么恢复到基础水平,要么在临床疾病期间直接逆转。引人注目的是,我们表明,这个簇包含一个高度类似于突触 N-甲基-D-天冬氨酸(NMDA)受体信号和神经保护途径激活的特征。此外,参与神经元投射和树突发育的基因也在整个疾病过程中发生改变,最终导致突触蛋白的基因表达普遍下降。同样,失调的 miRNAs,如 miR-132-3p、miR-124a-3p、miR-16-5p、miR-26a-5p、miR-29a-3p 和 miR-140-5p,也遵循伴随的表达模式。这是第一个深入的基因组研究,描述了海马神经元对早期朊病毒复制的临床前反应。我们的发现表明,朊病毒复制导致持续刺激程序性反应,至少部分由突触 NMDA 受体活性介导,该活性最初促进细胞存活和神经突重塑。然而,在感染的海马体中出现临床症状之前,这种反应就终止了,这似乎指向了疾病的一个关键转折点。对这些早期神经保护途径的操纵可能会改变退化和存活之间的平衡,为治疗提供潜在的途径。