Rohm Marlena, May Caroline, Marcus Katrin, Steinbach Simone, Theis Verena, Theiss Carsten, Matschke Veronika
Ruhr University Bochum, Medical Faculty, Institute of Anatomy, Department of Cytology, Bochum, Germany.
Ruhr University Bochum, Medical Faculty, Medizinisches Proteom-Center, Bochum, Germany.
Cell Physiol Biochem. 2019;52(6):1412-1426. doi: 10.33594/000000099.
BACKGROUND/AIMS: Amyotrophic lateral sclerosis (ALS) is the most common degenerative motor neuron disease in humans. However, the pathogenesis of ALS is not yet understood. The wobbler mouse is considered as an animal model for the sporadic form of ALS due to its spontaneous mutation in the Vps54 gene. Due to transactivation of NDRG2 by p53, this tumor suppressor might play a functional role in stress induced cell death in wobbler mice as well as ALS patients. Furthermore, deregulated microRNAs are often related to neurodegenerative diseases. Thus, the NDRG2 linked miR-375-3p was of interest for this study.
Here, we investigated the relevance of NDRG2 and miR-375-3p for the pathomechanism of the motor neuronal degeneration in wobbler mice by investigating expression level via qPCR and Western Blot as well as localization of these molecules in the cervical spinal cord by in situ hybridization, immunostaining and mass spectrometric analysis.
We were able to show a differential regulation of the expression of NDRG2 as well as miR-375-3p in the cervical part of the spinal cord of wobbler mice. In addition, for the first time we were able to demonstrate an expression of NDRG2 in motor neurons using different techniques.
The present study has shown NDRG2 and miR-375-3p to be promising targets for further research of the pathogenesis of sporadic ALS in the wobbler mouse model. Based on these results and in combination with previous published data we could develop a putative pro-apoptotic mechanism in the spinal cord of the wobbler mouse.
背景/目的：肌萎缩侧索硬化症（ALS）是人类最常见的退行性运动神经元疾病。然而，ALS的发病机制尚未明确。震颤小鼠由于其Vps54基因的自发突变，被认为是散发性ALS的动物模型。由于p53对NDRG2的反式激活作用，这种肿瘤抑制因子可能在震颤小鼠以及ALS患者的应激诱导细胞死亡中发挥功能性作用。此外，失调的微小RNA通常与神经退行性疾病有关。因此，与NDRG2相关的miR-375-3p成为本研究的关注点。
在此，我们通过qPCR和蛋白质免疫印迹法研究NDRG2和miR-375-3p的表达水平，并通过原位杂交、免疫染色和质谱分析研究这些分子在颈脊髓中的定位，以此来探究NDRG2和miR-375-3p与震颤小鼠运动神经元变性发病机制的相关性。
我们能够证明在震颤小鼠脊髓颈部，NDRG2以及miR-375-3p的表达存在差异调节。此外，我们首次使用不同技术在运动神经元中证实了NDRG2的表达。
本研究表明，NDRG2和miR-375-3p是进一步研究震颤小鼠模型中散发性ALS发病机制的有前景的靶点。基于这些结果并结合先前发表的数据，我们可以在震颤小鼠的脊髓中建立一种假定的促凋亡机制。
