Atxn2-CAG100-Knock-In Model for Spinocerebellar Ataxia Type 2

在由多聚谷氨酰胺结构域扩展引起的神经退行性疾病中，脊髓小脑共济失调2型（SCA 2）、肌萎缩性侧索硬化症（ALS 13）和左旋多巴反应性帕金森病据报道由突变型共济失调蛋白-2（RNA结合蛋白）触发。为了在小鼠中模拟这些疾病，在公开了42-谷氨酰胺-ATXN 2-敲入的表征之后，我们现在成功地产生了100-谷氨酰胺-ATXN 2-敲入。在这里，我们建议使用小脑和脑干/脊髓组织从老年小鼠阐明进行性神经退行性变，通过分子研究，相互作用分析，钙成像，电生理，组织学和行为测试。我们也计划建立小脑的器官型培养，以剖析关键分子的作用，并尝试拯救实验。总的来说，这种方法将记录扩增的ATXN 2对RNA和蛋白质的组织特异性作用，从而鉴定病理学的分子和功能生物标志物。

Among the neurodegenerative disorders caused by poly-glutamine domain expansions, Spinocerebellar Ataxia type 2 (SCA2), Amyotrophic Lateral Sclerosis (ALS13) and LevoDopa-responsive Parkinson were reported to be triggered by mutant Ataxin-2, a RNA-binding protein. To model these diseases in mouse, after the published characterization of a 42-glutamine-ATXN2-knock-in we now successfully generated a 100-glutamine-ATXN2-knock-in. Here, we propose to use the cerebellar and brainstem / spinal cord tissue from aged mice to elucidate the progressive neurodegeneration through molecular studies, interaction analyses, calcium-imaging, electrophysiology, histology and behaviour tests. We also plan to establish organotypic cultures of cerebellum to dissect the role of crucial molecules and attempt rescue experiments. Overall, this approach will document tissue-specific effects of expanded ATXN2 on RNAs and proteins, thus identifying molecular and functional biomarkers of pathology.